Resistin – possible immune regulator
I. Nagaev, M. Andersen, M. K. Olesen, O. Nagaeva, J. Wikberg, L. Mincheva-Nilsson and G. N. Andersen
In this paper by Scandinavian researchers it is shown that the inflammatory cytokine resistin (resistance to insulin) has pro-inflammatory as well as proresolving roles in active rheumatoid arthritis.
Rheumatoid arthritis is a debilitating autoimmune disease, which affects about 1% of the population in Western countries. Rheumatoid arthritis is T cell mediated, implying activation and expansion of auto-reactive T cells, their migration into the synovia and induction of local inflammation with monocyte activation, leading to cartilage damage and bone resorption.
In recent years, the cytokine resistin has attracted attention as a pro-inflammatory cytokine in rheumatoid arthritis.
The main finding presented in this paper is that resistin gene expression in CD4+ T cells is susceptible to changes in the cytokine environment, which indicate that resistin may have other properties than “just” being a pro-inflammatory cytokine secreted by monocytes. Considering this, the authors assume that resistin synthesis may be used by CD4+ T cells to direct the immune response into regulatory and/or effector Th1, Th2, Th17.
– Interestingly, we found a correlation between fold change in resistin and TGFβ gene expressions in monocytes, which points to a connection between the regulation of these two genes, says Marlene Andersen, first author of the paper.
Marlene shares the first authorship with Ivan Nagaev, who did all the biochemistry experiments in the study.
– I’m a medical doctor and Ph.D. student at Aalborg University and at present fulfilling my specialist training at North Denmark Regional Hospital to become a rheumatologist.
Furthermore, Marlene Andersen is a member of the “Scandinavian Melanocortin Study Group” of Grethe Neumann Andersen, with whom this study is done in collaboration.
– During the work that led to this paper, I have mostly enjoyed the rewarding cooperation with my amazing main supervisor, Grethe Neumann Andersen. She always shares her wisdom, knowledge and experience.
As TGFβ is important in the maintenance of immune tolerance, the findings presented here support the view that resistin may have an immune regulatory function.
– Our next step will be to study resistin gene expression in the context of a panel of cytokine gene expressions discriminating between Th1, Th2, Th17 and regulatory response in various leukocyte subsets from DMARD naïve patients with rheumatoid arthritis of recent onset, Marlene Andersen concludes.
On the HLA-DR-presence on T cells
A. L. S. Revenfeld, R. Steffensen, L. H. Pugholm, M. M. Jørgensen, A. Stensballe and K. Varming
Here, Danish researchers elaborate on the longstanding observation of HLA-DR being present on T cells.
It has been observed for more than four decades that T cells in peripheral blood can present MHC class II (MHCII) molecules on their outer surface and that the number of MHCII+ T cells increases upon activation. In line with this, one of the three human MHCII isotypes, called HLA-DR, is frequently used as a T cell activation marker along with other molecules, such as CD69 and CD25. Nonetheless, the functional role and significance of HLA-DR on human T cells is not fully determined and the unequivocal confirmation of an endogenous expression or a protein acquisition from other immune cells is absent.
The present study was carried out to elucidate the physiological details of HLA-DR on T cells as well as to obtain novel information about a possible endogenous expression.
Anne Louise Revenfeld, the first author of the paper, carried out the work that led to this publication as a part of her PhD, which she finalized about a year ago.
– The topic of my thesis was HLA-DR in relation to T cells and the article addresses some of the initial questions we wanted to answer. I think that the strength of our investigation is based on the combination of highly tailored protein and transcript analyses, which elucidate the phenomenon on different levels simultaneously
Especially one technique was very dear to Anne Louise during this work.
– My major interest was definitely centered on the flow cytometric work, as this is a fantastic and indispensible technique, when working with immunological research.
With her work, Anne Louise and her colleagues show that the characteristics for the presence of HLA-DR on T cells differ from when it is found on antigen-presenting cells.
– Since a lot of the expected functionality of HLA-DR on T cells has been deduced from what we know from its function on antigen-presenting cells, we question that this knowledge might simply be extrapolated to T cells. Hence, we do not immediately believe that HLA-DR-presenting T cells are antigen-presenting cells. In addition, the use of HLA-DR as an activation marker for T cells is questioned, and this is rather important because many laboratories still use HLA-DR in this context without relating their findings to something functional (e.g. WHY it is present on T cells and what does this mean for the pathological/physiological issues that are investigated?). With our results we hope that this practice may be re-evaluated.
Anne Louise Revenfeld is currently a postdoctoral researcher, doing mainly research about basic immunological mechanisms with a focus on CD4+ T cells.
– We continue to work with HLA-DR in the context of T cells during my postdoctoral work. Ultimately, it is the possible functionality of this phenomenon that we are interested in. Consequently, functional investigations have been initiated. We hope to show that HLA-DR is connected to regulatory functions of T cells and our work in this context also involves the role of extracellular vesicles produced by cells of the immune system, as means of communication and regulation.
C-type Lectin Receptors, Basophils and Allergen-Specific Immunotherapy
K. Lundberg, F. Rydnert, S. Broos, M. Andersson, L. Greiff and M. Lindstedt
In this recent publication, researchers from Lund University in Sweden show that several C-type lectin receptors are present on the surface of basophils, suggesting that basophils may play an important role for the body´s ability to recognize different type of agents and can regulate the immune system to a larger extent than was previously known.
Basophils are emerging as immunoregulatory cells capable of interacting with their environment not only via their characteristic IgE-mediated activation, but also in an IgE-independent manner. Basophils are known to express and respond to stimulation via different receptors such as TLR2, TLR4, DC-SIGN and DCIR, but whether basophils also express other C-type lectin receptors is largely unknown. In this study, the researchers investigate the C-type lectin expression profile of human basophils using multicolour flow cytometry.
Kristina Lundberg is one of the lead authors of the paper and she is a senior postdoc in the group. She participated in all the parts of the study.
– I actively took part in the designing of the study, and I and Dr Rydnert, the shared lead author were responsible for detailed planning, performance and analysis of the experiments, she says.
The presence of the C-type lectin receptors on basophils did change during so called allergen specific immunotherapy, implicating that they might play a roll in changing the immune response during this kind of treatment, leading to a lower allergic reaction.
– This study is the first to show a broader picture on how basophils express C-type lectin receptors, and it opens up for further investigations of the role of basophils during allergic reactions and other immune responses, which is very exciting, Kristina Lundberg concludes.
Cardiac Syndrome X and the complement system
Z. Horváth, D. Csuka, K. Vargova, S. Leé, L. Varga, P. Garred, I. Préda, E. T. Zsámboki, Z. Prohászka and R. G. Kiss
In this recent publication, researchers from Hungary show that complement system activation via the lectin pathway might play a role by inducing microvascular dysfunction in patients with Cardiac Syndrome X.
Patients with typical angina pectoris and inducible myocardial ischaemia, but with macroscopically healthy coronary arteries are commonly known as ‘cardiac syndrome X’. In these patients, a significantly elevated plasma level of terminal complement complex, the common end product of complement activation, has been observed without accompanying activation of the classical or the alternative pathways. Therefore, the aim of this study was to clarify the role of the ficolin–lectin pathway in Cardiac Syndrome X.
In summary, the researchers demonstrate significantly lower serum levels of lectin pathway parameters, namely ficolin-2, ficolin-3, ficolin-3/MASP-2 complex and FCN3-TCC deposition, and significantly higher terminal complement complex levels in patients with Cardiac Syndrome X compared to healthy controls or to patients with angiographically proven coronary heart disease.
Zsofia Horvath was a PhD student in the research group at the time of the study and is currently working as a clinician, a trainee in cardiology.
– Since this was a clinical study, I enrolled all the patient as well as collected and prepared the plasma samples, she says. Measurements were performed in the frame of an international collaboration. Data analysis and preparation of the first draft of the manuscript were performed in our research group.
Zsofia Horvath enjoyed the collaboration between the different research groups, and she is hopeful that another study in the same field will soon be published.
– We are doing analysis on acute and stable coronary heart disease patients and complement system activation via the alternative pathway and I also hope to be able to defend my thesis soon, she concludes.
On antiphospholipid antibodies in patients with systemic lupus erythematous
L. Garabet, I.-M. Gilboe, M.-C. Mowinckel, A. F. Jacobsen, T. E. Mollnes, P. M. Sandset and E.-M. Jacobsen
Here, researchers from Norway show that systemic lupus erythematous (SLE) patients with antiphospholipid (aPL) have significantly lower levels of complement C3 and C4 compared to SLE patients without aPL.
SLE is an autoimmune disease characterized by multiorgan involvement, presence of immune complexes in the affected organs and the excessive production of a variety of autoantibodies, including aPL antibodies. Activation of the complement system and low complement levels are common in SLE.
This study aimed to investigate the association between aPL and complement levels in patients with SLE.
Lamya Garabet is the first author of the paper:
– My main responsibility was, together with my supervisors, to plan the study, to do the required preparations as the ethical and data permission requirements before starting the study, acquiring the blood samples and the clinical information from the registry, putting the test results together and to do the statistical analysis together with my main supervisor Dr. Eva Jacobsen. Then I wrote the article, did the changes suggested by my co-authors and did the submission. The blood tests were analyzed by a study bioengineer who is a co-author; Marie-Christine Mowinckel.
– Our main findings are that SLE patients with aPL antibodies have significantly lower levels of complement C3 and C4 than SLE patients without aPL, and that there was no significant difference in CRP between aPL-positive and aPL-negative SLE patients. Our findings suggest that lower C3 and C4 values in aPL-positive SLE patients may reflect a higher consumption of C3 and C4 due to higher complement activation probably indicating that these antibodies cause complement-activation. Another possible explanation is that this subgroup of SLE patients has a higher disease activity and thus higher complement activation, Lamya Garabet concludes.
Association between type I IFN signature and a subgroup of myositis patients
L. Ekholm, S. Vosslamber, A. Tjärnlund, T. D. de Jong, Z. Betteridge, N. McHugh, L. Plestilova, M. Klein, L. Padyukov, A. E. Voskuyl, I. E. M. Bultink, D. Michiel Pegtel, C. P. Mavragani, M. K. Crow, J. Vencovsky, I. E. Lundberg and C. L. Verweij
In this study, Swedish, Dutch and Czech researchers reveal an association between the type I IFN signature and a subgroup of myositis patients with autoantibodies against RNA-binding proteins.
Myositis is a heterogeneous group of autoimmune diseases characterized by proximal muscle weakness and muscle inflammation, and can be subgrouped into polymyositis, dermatomyositis and inclusion body myositis. A common feature is the presence of autoantibodies.
The aim of the study was to test the hypothesis that autoantibodies directed against RNA-binding proteins in patients with myositis are associates with a type I IFN signature and thus potentially could induce IFN production.
Louise Ekholm, the first author of the paper is a PhD student at the Karolinska Institutet in the myositis group of professor Ingrid Lundberg.
– I will include this study in my thesis, which is defended on December 16 this year, she explains.
She has coordinated the work in this study, collected the clinical information on the myositis cohort as well as written the actual paper in collaboration with her co-authors.
– Otherwise, this project was collaboration between researchers here at the Karolinska Institutet, in the Netherlands and in the Czech Republic.
– Our study confirms that there are several different pathogenic mechanisms in the different sub groups of myositis. We have found that the IFN system is activated in patients with autoantibody mono-specific against RNA-binding proteins and in patients with multispecific antibody status, meaning that they have two ore more types of antibodies per individual.
The researchers also found that IFN alpha is mainly responsible for the IFN signature.
Taken together, the results from this study emphasize the need for careful molecular phenotyping of patients to gain better understanding of molecular pathogenesis and to improve treatment for myositis patients.
On the importance of the oligomeric state of MBL
T. R. Kjaer, L. Jensen, A. Hansen, R. Dani, J. C. Jensenius, J. Dobó, P. Gál and S. Thiel
In this publication, Danish researchers shed light on the importance of the oligomeric state of a lectin pathway pattern recognition molecule, MBL.
Troels R. Kjaer is the first author of the paper.
– I had recently finished my PhD and was working as a research assistant at time of preparation of this paper, he says.
Troels R. Kjaer was involved in all aspects of the work, e.g. designing, planning, and executing experiments, and in addition he had the main responsibility of writing the manuscript.
– When assessing complement activation and pathogen recognition, which are the classical features to explore for pattern recognition molecules, it is clear that higher oligomeric forms of MBL perform better. I believe that our main finding is the demonstration of this.
The group´s finding underscores that within the immune system it is too simplistic only to consider the one-to-one interaction between, e.g. two proteins. Instead it is important to consider the importance of multivalency that occurs in several situations.
– This publication is the outcome of fantastic cooperation between all parties involved. To be part of the pulsing research environment created by my co-authors has been a fun and rewarding experience. Obviously, the moment we observed results that correlated with our hypothesis was an exciting moment where things suddenly started to make sense.
MBL is just one of many pattern recognition molecules in the lectin pathway.
– It would be nice to see how oligomerization affects the others. Maybe the conclusions we draw here for MBL are transferable to these molecules, Troels R. Kjaer concludes.
On MBL in Hepatitis C virus infection
L. Zupin, V. Polesello, G. Alberi, G. Moratelli, S. L. Crocè, F. Masutti, G. Pozzato, S. Crovella and L. Segat
In this work, 203 Italian Hepatitis C patients and 61 healthy controls were enrolled and genotyped for the five MBL2 polymorphisms to investigate their role in Hepatitis C virus (HCV) infection susceptibility, spontaneous viral clearance and treatment response.
Hepatitis C is a common hepatic disease caused by infection with the hepatitis C virus, affecting globally around 130–150 million persons. An important component of the innate immune system, the mannose-binding protein C (MBL) potentially possesses a functional activity against HCV.
So, with the aim to investigate a possible role of MBL2 genetic variants in the context of HCV infection susceptibility, spontaneous viral clearance and response to certain therapies, an association study was performed for analysing MBL2 polymorphisms in a group of HCV patients and healthy controls from north-east Italy.
Additionally, a meta-analysis was performed to unravel the possible role of MBL2 genetic variants in HCV infection.
Although the MBL innate immunity molecule has been reported as possessing antiviral activity against HCV, in this study the researchers did not find association between functional MBL2 polymorphisms and susceptibility to be infected by HCV.
The genetic factors that influence the natural course of HCV infection and the response to treatment are not yet fully understood. In this work, MBL2 gene polymorphisms were analysed as a potential factor involved in HCV infection susceptibility and therapy outcome. MBL2 polymorphisms seemed to interfere marginally with interferon therapy response.
On immunosuppressive treatment during pregnancy
J. Kabat-Koperska, A. Kolasa-Wołosiuk, B. Wojciuk, I. Wojciechowska-Koszko, P. Roszkowska, B. Krasnodębska-Szponder, E. Paczkowska, K. Safranow, E. Gołembiewska, B. Machaliński and K. Ciechanowski
In this study, researchers from Poland show that there are qualitative, quantitative and morphological changes of the immune system in pharmacologically immunosuppressed female Wistar rats.
Female graft recipients are advised to become pregnant at least 1–2 years after successful kidney transplantation, when graft function is good and stable. In this period, a reduction in doses of immunosuppressive drugs is possible with acceptable low danger of acute rejection. Some immunosuppressive drugs are considered to be relatively safe during pregnancy while others are contraindicated due to their toxicity. However, experience regarding the use of many immunosuppressive drugs in human pregnancy is limited.
In this experimental study, the researchers assessed the impact of ‘safe’ and ‘contraindicated’ drugs in combinations (the ones most frequently used in therapy of human kidney recipients) on changes in the immune system of female Wistar rats after exposure during pregnancy.
Joanna Kabat-Koperska is the first author of this study and a senior assistant in the group.
– I designed this study, collected and interpreted the data and wrote the manuscript, she explains.
This publication is part of J.Kabat-Koperska´s scientific achievement that she prepared to apply for a habilitation degree in Poland.
Most previous studies focused on immunotoxicity were carried out on male rats, but here the group focus on female, pregnant rats, and find that thymus structure, spleen composition and splenocytes IL17 production were mostly affected in a drug regimen-dependent manner.
In summary, the researchers observed some important alterations in composition of splenic lymphocytes and their function on the basis of changed production of cytokines in female rats exposed to different combinations of immunosuppressive drugs during pregnancy. Altered sensitivity to immunotoxins or immune-modulating drugs can be expected in humans as well as the immune system of human changes during pregnancy, for example reflected by the frequent improvement of symptoms of rheumatoid arthritis.
Immature phenotype of regulatory T cells in neonates
C. Rennó, M. I. V. Nadaf, C. A. Zago, M. Carneiro-Sampaio andP. Palmeira
Here, Brazilian researchers have investigated the frequency of T regulatory cells (Tregs) in neonates and show differences in weight and gestational age among three groups of newborns (moderate and very preterm newborns, late preterm newborns and full-term newborns), which demonstrate the importance of a detailed division into groups.
Tolerance to self-antigens is a highly regulated process, and to maintain it, the immune system must be able to distinguish autoreactive lymphocytes and their development. Tregs are particularly important for the maintenance of immune homoeostasis, and failures in their development and function are the primary causes of a fatal autoimmune and inflammatory disease as evidenced in both humans and mice, which results from a mutation in the forkhead box P3 gene (FOXP3), the master transcription factor expressed in Treg.
The establishment of normal baseline values of Treg in healthy human newborns and how they are numerically and proportionately affected by prematurity provides the basis for further investigation into the function of these cells. Several authors have described reference values for leucocyte subpopulations in full-term neonates but few have analysed the same parameters in preterm newborns.
Camila Rennó was a Master’s degree student in Dr Patricia Palmeira´s lab and designed the overall study with her supervisor and collected samples at the Obstetric Center.
– I performed the experiments, analyzed and interpreted the data, and wrote the manuscript with Dr Patricia Palmeira, she explains.
The differences in weight and gestational age among the three groups of newborns shown in this publication can also be seen in the phenotyping results, such as the number of circulating leucocytes, which probably reflects the dynamic developmental phases, in which a difference of only a few days during pregnancy already shows a different degree of maturity.
Although higher frequencies of Treg cells are present in term and preterm newborns compared with adults and these are inversely correlated with gestational age, an immature phenotype with a higher expression of CD45RA and α4β7/α4β1 and a lower expression of CTLA-4 is found, particularly in the very preterm group.
– This could imply lower functional activity of these cells in neonates, particularly in the very preterm group, Camila Rennó concludes.
Bioinformatic screen for Ebola-vaccine candidates
T. Yasmin & A. H. M. Nurun Nabi
In a publication in the May issue of Scandinavian Journal of Immunology researcher from the University of Dhaka investigated potential candidates for designing a vaccine against Ebola virus. They performed sequence and structure based analyses and found a cluster of B cell and T cell specific epitopes as the best potential candidates for a vaccine.
Amongst the T cell epitopes, “LANETTQAL” and “FLYDRLAST” from glycoprotein wereconsidered as the most potent candidates because of their interaction with the highest number of different HLA alleles, conservancy, percentage of population coverage and docking analysis. On the other hand, peptides “79VPSATKRWGFRSGVPP9455” from GP1 and “515LHYWTTQDEGAAIGLA53056 ” from GP2 of Ebola glycoprotein were predicted as linear B cell epitopes of which the later contains a region 519TTQDEG524 that fulfilled all the criteria of accessibility, hydrophilicity, flexibility and beta turn region for becoming an ideal B cell epitope.
– We believe that this suggested cluster of B and T cell epitopes will definitely reduce time, cost and labor during in vivo and in vitro studies to be carried out for developing a vaccine against Ebola virus, says Tahirah Yasmin, first author of the study
Tahirah Yasmin is a lecturer at the Department of Biochemistry and Molecular Biology, University of Dhaka and a member in the research group supervised by Dr. A H M Nurun Nabi.
– I enjoyed discussing the study plan with my supervisor and witnessing how the plan evolved through these discussions, she says. But perhaps the most fun part was correlating these dry lab data with previous wet lab experiments conducted in some other parts of the world. This only shows the vast potential of bioinformatics in the current world.
Gum arabic have a potential pro-inflammatory effect
F. Bovo, R. M. Lenzi, F. T. Yamassaki, I. J. Messias-Reason, L. H. Campestrini, F. R. Stevan, S. F. Zawadzki-Baggio & J. B. B. Maurer*
In this study, researchers from Brazil demonstrate that gum arabic, considered to be an inert and widely used as an incipient in the pharmaceutical industry and as food additive, can induce activation of both alternative and classical pathway of complement system in vitro, and as such a potential pro-inflammatory effect.
This work was the result of a collaborative study of two research groups, in order to investigate of the effect of Gum arabic and cashew nut tree gum exudate polysaccharide (CNTG) on the classical and alternative pathway of the complement system.
Dr Bovo, the first author of the study is an employee of the Federal University of Parana (UFPR), and did her PhD and post-doctoral training in the research group supervised by Prof. Dr. Iara de Messias Reason (Laboratory of Molecular Immunopathology) and by Prof. Dr. Juliana Maurer (Parana Center for Research in Medicinal Plants- NUPPLAMED).
– Doing research in the area of immunology specifically on complement system has always made me very happy, she says.
Ever since her first year of college she was interested in immunology and thus enjoys doing this research. She points out the importance of a good supervisor.
– Working under a good supervisor that always shares her wisdom, knowledge and experience, having a good work team and achieving significant results is really fulfilling, Dr Bovo says.
However, there are some drawbacks to her days:
– Our tasks as researchers are quite difficult to fulfill most of the time in our country due to financial restrictions and a lack of official support, she concludes.
Ageing and lymphocyte subsets
R. Valiathan, M. Ashman and D. Asthana
Here, researchers from the University of Miami Miller School of Medicine retrospectively analysed the data for complete blood count (CBC) and lymphocyte subsets from infant to elderly age groups (infants, children, adolescents, adults and elderly) to determine changes taking place during ageing.
The researchers show that B cells continuously decrease from childhood, which highlights the importance of establishing age specific reference points.
Ranjini Valiathan is a postdoctoral associate in the group, and was responsible for the lab work, analysis and writing of the article.
– The NK cells represent an important component of the innate immune response against infection and we observed a significant decrease in NK cell percentages from infancy to adolescent and then an increase from adults to the elderly population, she explains.
In addition, the researchers investigated levels of epidermal growth factor, EGF, involved in the growth and proliferation of several types of cells in the central nervous system.
– In our analysis there were decreased plasma EGF levels in elderly individuals compared to adults, Ranjini Valiathan says. High levels of EGF are present in the CNS and it plays a crucial role in controlling proliferation and differentiation of nervous tissue during neurogenesis. Additionally, it is also a major factor in promoting wound healing by expression at sites of injury, which inhibits the activity of nitric oxide synthase, preventing inflammation.
– This study revealed the changes in many different lymphocyte subsets and blood count parameters during ageing process and shows us the requirement of establishing age specific reference points, she concludes.
On obesity and inflammation
P. Ramos-Ramírez, C. Malmhäll, K. Johansson, J. Lötvall and A. Bossios
In a study by researchers from the Krefting Research Center at the University of Gothenburg it is found that T-regulatory cells in adipose tissue express Helios, a transcription factor that it has been suggested as a marker for thymic-derived T-regulatory cells.
Obesity has increased significantly in recent decades and has become the largest global chronic health problem in adults. It is now recognized that in addition to its role as a storage depot for lipids, adipose tissue also secretes diverse proteins that influence metabolism as well as the immune system.
The aim of the present study was to evaluate the adiponectin receptor 1 (AdipoR1) expression in adipose tissue-resident T-regulatory cells and to evaluate the effect of weight gain on this expression.
The researchers found that adipose tissue-resident T-regulatory cells express high levels of AdipoR1 compared with their circulating counterparts in the spleen, suggesting an adipose tissue-specific function of AdipoR1-positive T-regulatory cells.
Patricia Ramos-Ramirez, originating from Mexico came to Sweden to do a postdoc at the Krefting Research Center, in the group of Dr Apostolos Bossios, associate professor at the University of Gothenburg.
– In this work, I was involved every aspect, from the experimental design and maneuverers, such as tracking weight gain of the mice, the collection and processing of samples, analysis of data and of preparing the manuscript, she says.
Patricia Ramos-Ramirez enjoyed the data analysis of the research very much.
– After seeing tiny cells doing, or not, their work, you can understand some of the mechanisms leading to the development of pathologies, as in this case the obesity, she says. The most fun during a project is when you find a possible answer to your initial question, but often you find an answer and also more questions, and it is very exciting to generate new hypotheses and ways to understand how cells work in homeostasis or during pathologies, as in our study, during obesity, she concludes.
Obesity is a growing public health problem, and understanding as the low-grade inflammation during obesity is regulated could be a potential subject to control it.
Cytokine profile in patients with DiGeorge syndrome
D. M. Aresvik, K. Lima, T. Øverland, T. E. Mollnes and T. G. Abrahamsen
In a study conducted by Norwegian researchers an association between increased levels of the pro-inflammatory and angiostatin chemokine IP-10 and presence of congenital heart disease in patients with the 22q11.2 deletion syndrome is shown.
The syndrome, also known as DiGeorge syndrome, is a genetic disorder with an estimated incidence of 1:4000 births. Patients may suffer from affection of many organ systems with cardiac malformation, thymic hypoplasia or aplasia, hypoparathyroidism, palate anomalies and psychiatric disorders being the most frequent. The incidence of autoimmune diseases is increased in older patients.
In the present study, researchers examined a cytokine profile in patients with DiGeorge syndrome by measuring a broad spectrum of serum cytokines.
Dina Aresvik is the first author of the paper, and a PhD student in the group of Tore Abrahamsen. She planned the study with her supervisor and their collaboration partnerTom Eirik Mollnes. In addition she performed the laboratory work and analyzed the data.
– I just love the feeling you get when you discover something in the lab, she says, however admitting that the favorite part of the study was working with the patients.
– Many children participated in our study, and it was very meaningful to work with them.
Dina Aresvik did also write the paper in collaboration with the rest of he research group, and they conclude that their finding that the pro-inflammatory and angiostatic chemokine IP-10 is increased in DiGeorge-patients may have implications for the pathogenesis of the syndrome. However, further studies are needed to clarify the association between increased IP-10 and congenital heart defects.
Activation of Complement Following Total Hip Replacement
S. Thordardottir, T. Vikingsdottir, H. Bjarnadottir, H. Jonsson Jr and B. Gudbjornsson
Here, researchers from Iceland show that there is significant activation of the complement system following cemented total hip replacement surgery. The study also indicates that these processes are likely still active on the 3rd post-operative day.
First author of the study, Soley Thordardottir, was doing her bachelor internship at the Landspitali – University Hospital in Iceland under the supervision of Thora Vikingsdottir and Bjorn Gudbjornsson.
– Although during the time the article was submitted, revised and accepted for publication, I was a PhD student at the Radboud university medical center in the Netherlands, she says.
Soley Thordardottir performed the majority of the experimental work, analysed the data, conducted statistical analysis and prepared the manuscript for publication.
– What I found most fun was to go and pick up the freshly drawn blood samples from the orthopaedic unit, because there I got to witness the cooperation and willingness of the patients involved in the study. It was inspiring to hear their curiosity and excitement about the study results, encouraging my believes that findings of studies like these might in the future help speed up recovery of patients and reduce the chance of painful complications following total hip replacement or other joint replacement surgeries.
Previously, the group have demonstrated that patients undergoing total hip replacement due to osteoarthritis develop an acute inflammatory reaction measured by an increase in CRP and cytokine levels as well as a decrease in albumin serum levels. Here, they investigated whether the complement system was also activated in these patients and show that the complement system indeed is activated for a longer period and conclude that this activation may play a role in the inflammatory reaction following the surgery.
The perks of papain-based vaccination
N. Abdel Aziz, H. Tallima, E. A. Hafez and R. El Ridi
Here, Egyptian researchers show that injection of mice with papain one day before exposure to S. mansoni appeared to diminish or shut off all infection-induced epidermal and lymph node cytokine signals, which were relegated to levels similar to or lower than those released by naive mice.
Schistosoma mansoni and Schistosoma haematobium infect more that 200 million people, causing severe disease characterized by intense inflammatory responses to the parasite eggs that fail to exit the mammalian host to continue the life cycle in the intermediate snail stage.
Nada Abdel Aziz was doing her masters under supervision of Associate Prof. Hatem Tallima, Prof. Ebtisam Hafez and Prof. Rashika El Ridi
– My responsibilities were doing the practical work and analyzing the data under my supervisors’supervision and monitoring the mice.
In the present study, the researchers started to elucidate the molecular basis of C57BL/6 mouse resistance to S. mansoni following treatment with the cysteine peptidase papain. They attempted to detect the early immunological signals in the epidermal, dermal and draining lymph nodes following S. mansoni infection in untreated and papain-pretreated mice.
– The main findings here are that Schistosoma mansoni infection in C57BL/6 mice induces immunological chaos and that a papain pretreatment or papain-based vaccination diminishes or shuts off S. mansoni infection early induction of type 1, type 17, and type 2 cytokines except for thymic stromal lymphopoietin, and respectively, re-programs or conditions the immune system towards a polarized type 2 immune milieu, Nada Abdel Aziz says.
Similarly to other helminthic parasites, significant resistance to S.mansoni infection is dependent on type 2 immune responses.
The researchers believe that the establishment of infection can be prevented if a proper composition of a cysteine peptidase-based vaccine can be selected, that is capable of entirely overcoming the infection-induced immunological chaos and generating a powerful, schistosome ESP-specific type 2 polarized memory immune response.
– The most fun during this study was that we strongly believe and are sure that our work will have a great impact on people’s life, particularly those 800 million who are at risk of infection which is the main reason actually, that creates high inspiration and motivation to do such work, Nada Abdel Aziz concludes.
Molecular mechanisms of macrophage M1/M2 polarization
L. V. Sakhno, E. Ya. Shevela, M. A. Tikhonova, A. A. Ostanin and E. R. Chernykh
In this study performed by Russian researchers it is demonstrated that GM-CSF-induced macrophages are not stringently predetermined to M1 polarization.
Macrophages are highly plastic cells which can differentiate into either a classical, pro-inflammatory M1 type or an alternatively activated, anti-inflammatory M2 type. Activation conditions determine which type they differentiate into. The present study was aimed at the investigation of cytokine production and pro-apoptogenic/inhibitory molecule expression in macrophages generated with GM-CSF using either different conditions (with or without serum and different growth factors).
The researchers find that deprivation of serum factors in the cultures of GM-CSF-differentiated macrophages skew their phenotype towards M2-like cells and that macrophages generated in growth factor deficient conditions differ from conventional M1 cells by the decreased expression of HLA-DR molecules, higher expression of M2-marker CD206 and pro-apoptogenic/co-inhibitory molecules TRAIL, FasL, B7-H1 and also possessed a decreased ability to stimulate the proliferation of allogenic PBMCs.
In summary, the study expands the knowledge of the molecular mechanisms of M1/M2 polarization.
Association between childhood asthma and the expression of functional markers by mDCs
H. Kääriö, J. K. Nieminen, A. M. Karvonen, K. Huttunen, P. C. Schröder, O. Vaarala, E.
In this study, a group of Finnish researchers for the first time describe phenotype and functional properties of circulating dendritic cells in association with asthma and farm exposure at 4.5 year old children.
The aim of the investigation was to explore where asthma and farm exposures are associate with the proportions and functional properties of dendritic cells from 4.5 year-old children in a subgroup of the Finnish PASTURE birth cohort study.
Farm exposure in early childhood is known to protect from childhood asthma and allergies, suggesting effects on the maturing immune system. Here, the researchers suggest an association between childhood asthma and the expression of functional markers by myeloid dendritic cells. Farm exposure may have immunomodulatory effects by decreasing mDC proportions.
Heidi Kääriö was a PhD student in the group during the course of this study.
– My main responsibilities during this study were laboratory work, data analysis and writing the manuscript, so basically all but sample collection, she says.
Heidi Kääriö defended her thesis, where this paper is included, in December last year.
– Thinking about this study this as a part of my thesis the main finding was definitely the lower myeloid dendritic cell-proportions of farm children when compared to non-farm children, she says.
Heidi Kääriö enjoyed designing the laboratory analyses.
– But later, when I was writing my thesis, the most fun part was to realize that this publication is really part of the bigger picture going on here.
All in all, this study provides important insights for the future studies investigating underlying immunological mechanisms of farm-related asthma-protection.
Reference genes for qPCR
D. B. Oturai, H. B. Søndergaard, L. Börnsen, F. Sellebjerg and J. Romme Christensen
Here, a group of researchers from Copenhagen, Denmark, have identified suitable reference genes for quantitative real-time PCR studies using different peripheral blood cell subsets from patents with relapsing-remitting multiple sclerosis, interferon-beta-treated patiens with relapsing-remitting multiple sclerosis and healthy controls.
Determination of gene expression in different cells or tissues using quantitative real-time PCR is today an essential method in understanding many biological and pathological mechanisms. The application of a reliable reference gene (constitutively active, endogenous gene) is a prerequisite to obtain correct estimations of target gene expression.
Jeppe Romme Christensen, MD, PhD is a post doc at the Department of Neurology at Rigshospitalet, Copenhagen University Hospital and he conceptualized and designed the study and supervised the first author.
– I think our study demonstrates that reference gene expression and stability varies substantially between different peripheral blood subsets, which have important implications for gene expression studies. Furthermore, I think our study highlights that the widespread use of GAPDH as single reference gene should be abandoned.
Taken together, the findings demonstrate that PBMC subsets and interferon-beta contribute substantially to reference gene stability and recommend the use of specific reference genes, single or in combination, for future studies involving patients with MS, PBMC subsets and interferon-beta treatment. The use of GAPDH as reference gene is not recommended, as it was generally the least stable reference gene.
New ways of targeting antigen to dendritic cells
L. H. Pugholm, L. R. Petersen, E. K. L. Søndergaard, K. Varming and R. Agger
The December issue of Scandinavian Journal of Immunology contains two papers from a Danish group at Aalborg University, covering different aspects of the same topic – namely targeting of antigens to dendritic cells. The first paper comprise an in vivotargeting study investigating a panel of different targets. The second paper describes an in vitro targeting assay that confirmed the results obtained in the in vivo targeting study, which enable this assay to be chosen as at first-line evaluation of potential targets.
Lotte Hatting Pugholm was a PhD student at Laboratory of Immunology, Aalborg University and at Department of Clinical Immunology, Aalborg University Hospital at the time these studies were undertaken. Now she is a post doc in a research group at the Department of Clinical Immunology at the Aalborg University Hospital.
– The work was done during my PhD, thus I was responsible for planning and performing all or the majority of the work related to the data, she says.
In addition, Lotte Hatting Pugholm drafted the manuscripts but the study design was a joint decision.
The main findings in the in vivo study was that targeting of antigen to dendritic cells can induce strong humoral responses against an otherwise weak antigen, even without addition of adjuvant and that inherent properties of the targeting antibodies are of great importance for the outcome. The in vitro study demonstrated that it is possible to obtain comparable results by the presented in vitro screening assay.
Lotte Hatting Pugholm like the work that led to these publication
– I especially enjoyed working with laboratory mice. In addition, setting up the ELISA test for investigation of the humoral response was interesting. The fact that we looked for mouse IgG antibodies directed against rat IgG made the ELISA setup more complicated than usual and needed several steps of optimization, she concludes.
Investigations on the role of FURIN in plasma
N. Ranta, H. Turpeinen, A. Oksanen, S. Hämäläinen, R. Huttunen, R. Uusitalo-Seppälä, E. Rintala, J. Aittoniemi and M. Pesu
In a prospective cohort study, performed by researchers from University of Tampere, FURIN levels in plasma from 537 patients was determined using ELISA. The patients were admitted to the emergency room with suspected infection.
FURIN belongs to the family of proprotein convertases, is widely expressed and regulates the maturation of numerous precursor protein such as growth factors, enzymes, hormones, cytokines and much more. In addition, it is also involved in the pathogenesis of several diseases.
The group in Finland have previously shown that FURIN is upregulated upon the activation of both T lymphocytes and innate immune cells, and others have demonstrated many pathogens are activated through FURIN/PCSK dependent proteolysis. However, whether the FURIN levels in plasma are also upregulated upon immune activation had not previously been studied.
– We hypothesized that patients with infections might have unregulated plasma FURIN levels, but we found that, at least with the method we used, elevated plasma FURIN does not associate with infections or predict patient survival, says Marko Pesu, principal investigator and Associate Professor at the University of Tampere.
In contrast, patients that had rheumatic disease had significantly more often elevated plasma levels of FURIN. Thus, it might be that plasma FURIN is not a good marker for infections, but it could be beneficial in the diagnostics of autoimmune diseases.
– Of course more work is needed, because the patient cohort we used was not collected to study autoimmunity, says Marko Pesu who was also not very happy about the sensitivity of the FURIN measurement.
Nevertheless, Marko Pesu was glad to see that observations from basic research, i.e. FURIN up-regulation inside immune cells might be beneficial for future diagnostics in rheumatoid diseases.
– It was also fruitful to collaborate with doctors who work with infectious patients.
In the future, Marko Pesu and his group would like to continue evaluation plasma FURIN levels in different patient cohorts, especially those with autoimmunity, given a more sensitive method for FURIN detection is available.
Dynamics of tissue redistribution of human NK cell subsets
Tissue Distribution Dynamics of Human NK Cells Inferred from Peripheral Blood Depletion Kinetics after Sphingosine-1-Phosphate Receptor Blockade
M. Mehling, A.-V. Burgener, V. Brinkmann, G. R. Bantug, S. Dimeloe, G. Hoenger, L. Kappos and C. Hess
In this study, conducted by researchers in Switzerland, the dynamics of tissue redistribution of human NK cell subsets in a cohort of patients with multiple sclerosis newly treated with the drug fingolimod was investigated.
In humans, two main functional subsets, CD56bright and CD56dim are distinguished. They both originate from the bone marrow, but following egress they redistribute in a tissue-specific manner. The dynamics with which these NK cell subsets redistribute to tissues remain unexplored. Here, the researchers took advantage of the mode of action of the drug fingolimod which efficiently blocks spingosine-1-phosphate-directed lymphocyte egress from tissue to blood. By studying depletion kinetics of NK cells in the blood of de novo fingolimod-exposed individuals, the researchers monitored indirectly dynamics of the redistribution of CD56bright and CD56dim NK cells from blood to tissue.
During this project the shared first co-authors M. Mehling and AV. Burgener worked at the Immunobiology research group of the Department of Biomedicine (Basel/Switzerland) as a postdoc and a master student, respectively. M Mehling also works as a neurologist at the University Hospital of Basel.
M. Mehling was involved in conceptualizing the study and organized most clinical and laboratory-related study logistics. Together, M. Mehling and AV. Burgener performed the experimental work, analysed the data, and conducted statistical analysis. Both were significantly involved in preparing the manuscript.
– For us, the main findings were the differential depletion kinetics of CD56bright NK cells and CD56dim NK cells cells following the first dose of fingolimod as this indicates a differential tissue distribution of these NK cell subpopulations. Specifically, this indicates that CD56bright NK cells cells preferentially home to secondary lymphoid organs where they are trapped by fingolimod, whereas CD56dim NK cells distribute mostly to non-lymphoid tissue.
In all, these data add basic insight to the understanding of the tissue distribution dynamics of NK cell in humans and by this to the immune surveillance process.
The researchers enjoyed this project that can have an impact on patients.
– The most fascinating moment was when we realized that we were able to capture with our approach differential depletion kinetics in NK cell subsets following the first dose of fingolimod. It was truly exciting to see that our hypothesis was correct and that we could generate biologically meaningful data while immunomonitoring humans starting with this selective immunomodulatory therapy.
New insights into the regulation of BCR signalling
IRF4 Deficiency Leads to Altered BCR Signalling Revealed by Enhanced PI3K Pathway, Decreased SHIP Expression and Defected Cytoskeletal Responses
P. M. Budzyńska, M. Niemelä, A.V. Sarapulov, M.K. Kyläniemi, K.-P. Nera, S. Junttila, A. Laiho, P.K. Mattila, J. Alinikula and O. Lassila
Here, a group from Finland show that transcription factor interferon regulator factor 4 (IRF4) has an important role in the regulation of BCR signalling and help to shed light on the molecular mechanisms of B cell development and germinal centre response.
Graded expression of IRF4 regulates B cell development and is critical for plasma cell differentiation. Here, researchers have characterized molecular targets of IRF4 in B cells, by establishing an IRF4-deficient cell line.
Paulina Budzynska is a PhD student in the research group of Professor Olli Lassila at the Department of Medical Microbiology and Immunology at the University of Turku. This article is a part of her PhD thesis.
– When I joined Olli Lassila’s laboratory I started this project by writing a study plan, I designed and performed most of the experiments, analysed the obtained data and wrote the first draft of the manuscript, she says. Furthermore, I was responsible for conducting active and fruitful collaboration with other authors especially with Dr Pieta Mattila whose research expertise is in the lymphocyte cytoskeleton and advanced microscopy.
Using the IRF4-deficient cell line Paulina Budzynska and co-workers found a compromised B cell receptor signalling in the absence of IRF4. In more detail, IRF4 disruption upregulated the BCAP and downregulated SHIP expression, leading to strongly enhanced activity of PI3K/Akt pathway upon BCR ligation. It was also found that attenuated early BCR signalling events in stimulated IRF4-deficient cells severely compromised signalling to the actin cytoskeleton.
– I think that this study advances our knowledge about the transcriptional role of IRF4 in the regulation of BCR signalling and helps to shed light on the molecular mechanisms of B cell development and activation response, since in physiological conditions vast majority of germinal center B cells lack IRF4 expression, Paulina Budzynska says. In addition, the study provides insights into how B cells might sense the antigen in germinal centers.
Paulina Budzynska appreciate spending time in the lab.
– I really enjoyed performing the experiments in this study. The greatest fun was waiting in the dark room for results from Western blots. It was so exciting to be the first to see the unknown, she concludes.
New link between TL1A and pro-inflammatory cytokines in inflammatory bowel disease
T. J. Ślebioda, A. Bojarska-Junak, M. Stanisławowski, M. Cyman, P. M. Wierzbicki, J. Roliński, K. Celiński and Z. Kmieć
In a study in the October issue of Scandinavian Journal of immunology a group of researchers from Poland show that in humans the tumor necrosis factor superfamily member TL1A may contribute to intestinal inflammation by local induction of expression of pro-inflammatory cytokine IL-17A. They also show that TL1A and its receptor DR3 is expressed not only on mononuclear cells present in intestinal lamina propria but also on enterocytes.
– The role of TL1A in inflammatory bowel disease is very complex and still remains not entirely explained, says dr Tomasz Slebioda, post-doctoral researcher at the Department of Histology at the Medical University of Gdansk.
The present study aimed to establish a link between expression of TL1A and selected TL1A-induced pro-inflammatory cytokines involved in the pathogenesis of inflammatory bowel disease. In addition, the researchers wanted to investigate a connection between serum concentration of TL1A in patients with inflammatory bowel disease and activation of peripheral blood T cells.
Tomasz Slebioda designed the study and conducted most of the research and especially enjoyed the ability to continue and expand the studies which he started as a PhD student at the University of Southampton (UK) under the supervision and direction of Prof. Aymen Al-Shamkhani.
– It has to be emphasized, however, that this publication is a result of successful cooperation with a team of medical doctors and scientists from Medical University of Lublin in Poland, he says.
In summary, the data presented in this publication indicate that TL1A may contribute to pathogenesis of inflammatory bowel diseases via local but not systemic induction of IL-17A but not IL-4, IL-13 or IFN-gamma.
Novel mechanism by which Varicella-zoster virus suppresses the antiviral response of the host
E.-J. Choi, C.-H. Lee and O. S. Shin
Here, a group of Korean researchers demonstrate that the expression of suppressor of cytokine signaling (SOCS)3 impaired host control of Varicella-zoster virus, resulting in modulation of type I IFN signaling and viral replication.
Varicella-zoster virus is an important viral pathogen that is responsible for causing varicella (chickenpox) hand herpes zoster (shingles). The virus has been shown to suppress early anti-viral innate immune responses, but the exact mechanisms are not yet well understood.
– In this study, we used three different cell types, keratinocytes, fibroblasts, and macrophages to investigate VZV-induced anti-viral responses, Eun-Jin Choi says.
Eun-Jin Choi is the first author of the study and is a post doc in the lab of infection and immunology at Korea University.
– It was intriguing to observe distinct characteristics of immune responses against Varicella-zoster virus infection, depending on the cell types, she says.
Taken together, the data presented in this publication indicate that Varicella-zoster virus-induced expression of SOCS3 represents a viral evasion mechanism and suggest that type I IFN signaling may be modulated by SOCS3-mediated immune suppression. This is a novel mechanism by which Varicella-zoster virus suppresses the anti-viral response of the host and paves a path to efficient virus replication.
Previously unknown role for IRF4 in liver transplant rejection
W. Zhao, Z. Zhang, Q. Zhao, M. Liu and Y. Wang
In this study, published in the September issue of Scandinavian Journal of Immunology, a Chinese research group shows that interferon regulatory factor 4 (IRF4) plays a crucial rule in regulating acute liver allograft rejection.
Acute liver allograft rejection is a serious complication after liver transplantation. IRF4 is expressed predominantly in the immune system and plays an important role in its development and function. However the role of IRF4 in liver transplantation has not been investigated before.
Wei Zhao is the first author of this paper and was in charge of experiments, analysis and interpretation of the data.
– In this study, we show that inhibition of IRF4 attenuates acute liver allograft rejection in mice, she says.
This effect of IRF4 is associated with promoted M2 macrophage differentiation; the study show an up regulation of the M2 numbers in liver of recipients treated with IRF4 siRNA.
The results in this study suggest that IRF4-inhibition reprogrammed macrophages to the M2, thereby favoring a ”graft protecting” status rather than progressive inflammation and destruction. IRF4 siRNA treatment increases IL-10 production and promotes M2 macrophage differentiation both in vitro and in vivo.
Limited cutaneous SSc and diffuse cutaneous SSc – more than two sites of the same coin
A. Wagner, M. Köhm, A. Nordin, E. Svenungsson, J.M. Pfeilschifter and H. H. Radeke
In this study performed by German and Swedish researchers it is shown that the soluble receptor for IL-33, sST2 is elevated in late phase limited cutaneous systemic sclerosis (lcSSc) as compared to patients with shorter disease duration or with the diffuse subtype of SSc. The researchers demonstrate that sST2 and not IL-33 is significantly increased in serum of lcSSc patients with disease duration over 9 years.
Annika Wagner was a PhD in the group of Professor Radeke when this study was undertaken and was responsible for the recruitment of the German patients and control subjects. She also prepared major part of serum and plasma samples, performed the ELISA measurements and did some of the statistical analyses.
– Our results give an additional hint towards the hypothesis that limited cutaneous SSc and diffuse cutaneous SSc seem to be more than two sites of the same coin. The ongoing discussion if lcSSc and dcSSc are two complete different diseases may give us the opportunity to gain a deeper understanding of the pathophysiology of lcSSc and dcSSc. This might result in an earlier diagnosis of the “two forms of SSc” and may give the physicians new opportunities to improve SSc treatment.
Data raised within this trial highlight the functional interaction of IL-33 and its neutralizing receptor sST2 in pathological processes of SSc and stresses the role of sST2 as a progression marker of lcSSc.
Annika Wagner, who is a trained biologist appreciated the work with this study and was happy to get a deeper insight into clinical research.
– And I got the chance to learn capillaroscopy at a workshop by the capillarscopy expert professor Cutolo, that was amazing, she concludes.
No difference in vitamin D levels in HIV patients and healthy controls
C. Missailidis, J. Höijer, M. Johansson, L. Ekström, G. Bratt, B. Hejdeman and P. Bergman
In this study, performed by researchers at Karolinska Institutet, it was found that vitamin D levels do not differ between chronic HIV patients and healthy controls. The researchers also found that despite perfect viral control, the HIV patients in the study still express higher levels of biomarkers of systemic inflammation than the control group. Vitamin D levels do not associate with this higher degree of systemic inflammation.
Catharina Missailidis is a PhD student in the group of Peter Bergman and she took an active part in all aspects of this study, from formulating study plan, objectives and ethical proposal, collecting samples, performing some of the assays, analysing data and drafting the manuscript.
– I thoroughly enjoyed all aspects of the study from hypothesis to written presentation.
Primary HIV infection causes massive damage to lymphoid and mucosal tissues leading to microbial translocation across a defective gut mucosa and progressive immunodeficiency. The overall aim of the current study was to provide baseline data for a future interventional trial testing the hypothesis that vitamin D could reduce this microbial translocation. However, the results from this study do not support such a study, where vitamin D is given to chronic HIV patients in Sweden.
Immunoinformatic design of new Ebola vaccine
M. A. Khan, M. U. Hossain, S. M. Rakib-Uz-Zaman and M. N. Morshed
Researchers from Bangladesh here use immunoinformatics to design an epitope-based vaccine against the Ebola virus. In addition, the group also performed a genome wide search to recognize the most suitable drug target site and simulated inhibition of the target site by a predicted inhibitor molecule.
Arif Khan, is a M.Phil student in professor Mohammad Neaz Morshed´s Chemoinformatics research group. He worked with conception and design of the study, performed the immunoinformatics, acquisition, analysis and interpretation of data, molecular docking study and drafted the manuscript.
– Active and fruitful collaboration with the other authors especially professor Mohammad Neaz Morshed led to this successful publication, Arif Kahn says.
Many Ebola vaccine candidates have been developed over the last ten years, but none has yet been approved for clinical use in humans. The aim of this study was to recognize major immunogenic epitopes on Ebola viral proteins and predict a vaccine as well as effective inhibitor binding sites. This study will enhance future laboratory-based attempts to develop effective treatment and prevention of Ebola virus infection.
T cell activation status in diabetic patients after dialysis
A. Almeida, O. Lourenço and A. M. Fonseca
In a study performed by researchers in Portugal it is shown that a haemodialysis session in patients with diabetic nephropathy affects their T cell activation status in the two major subpopulations and differentially modulates the production of inflammatory cytokines.
Dr Ana Mafalda Fonseca is a post doc in the research subgroup Human Integrative Immunology at the Health Sciences Research Centre at the University of Beira Interior in Covilhã, Portugal and coordinated the study.
– Lab work with human samples needs a dedicated and organized team and all steps contribute to the final work, she says.
The objective of the study was to assess the expression of activation-related markers on T cells, as well as to quantify inflammatory cytokines, before and after a single haemodialysis session in patients with diabetic nephropathy. A group of 17 patients where included in the study and the effect of haemodialysis on T cells and their major subpopulations, CD4 and CD8 was performed.
It is known that a significant proportion of patients with diabetes develop diabetic nephropathy. Dialysis causes some relevant changes in the immune system such as the activation of complement, the functions of monocyte-derived dendritic cells and their release of various pro-inflammatory cytokines. This study has focused on the effect of dialysis on T cells, specifically looking at changes that occur after a single haemodialysis session in the same individual. The Portuguese researchers find here a significant reduction of CD8 T cells after dialysis, but also an increase in the T CD4 subset, which promotes a continuous state of activation of T cells.
– With all the improvement in the haemodyialysis equipment, such as the dialyzer membranes, I didn´t expect the impact that the procedure has on the T cells activation status and inflammation. So, the immunomodulatory effects that haemodialysis sessions have on patients still impresses me, Ana Mafalda Fonseca says.
New insights into the role of B10 cells in Wiskott-Aldrich Syndrome
H.-Q. Du, X. Zhang, Y.-F. An, Y. Ding and X.-D. Zhao
In this paper a group from China show that Wiskott-Aldrich Syndrome protein (WASp) deficiency adversely affects the homeostasis of B10 cells in both humans and mice. This may cause an imbalance in the frequency of Th1 cells and regulatory T cells, resulting in a pro-inflammatory environment.
WASp is an important regulator of the actin cytoskeleton and is required for immune cell function. A mutation in the WAS gene causes Wiskott-Aldrich syndrome (WAS), an X-linked primary immunodeficiency disease. WASp deficiency leads to a reduction in the number of transitional type 2 marginal zone precursor and marginal zone B cells.
In this paper the group set out to investigate if WASp deficiency also lead to a reduction of regulatory B cells, known as B10 cells, a reduction that could contribute to the autoimmunity of WAS-patients. Looking both at patients and WASp knock out mice they found that the percentage of B10 cells was reduced in both mice and patients.
Hong-qiang Du is a master degree candidate in the group of professor Zhao and the first author of this paper.
–When looking at B10 cell frequency we found that they are decreased in many tissue compartments, indicating a global effect of WASp deficiency on B10 cells, which is very striking he says.
In the mouse experiments the group used old WASp KO mice with spontaneous autoimmune colitis instead of WASp KO mice with induced inflammatory status and found an imbalance of Th1/Treg ratio, possibly due to defective B10 cells,
– This is another important finding I think, Hong-qiang Du says.
Autoimmunity in Wiskott-Aldrich syndrome has long been an intriguing field of research.
– I believe that there are missing parts in the picture of cellular mechanism underneath this phenomenon, Hong-qiang Du says.
During the design of the research and writing of the manuscript Hong-qiang Du got a comprehensive and detailed understanding of autoimmune disease and autoimmunity in WAS, leading him towards his master degree.
Cytokines and chemokines in the pathogenesis of malaria during pregnancy
R. Megnekou, A. Lissom, J. D. Bigoga and J. C. Djontu
Here a Cameroonian group of scientists has studied this effects of pregnancy-associated malaria on Th1, IL-10 family Th17 cytokines and on CXCL-10 chemokine profiles in pregnant woman. They find that during pregnancy-associated malaria the IFN-gamma inducible protein 10, CXCL-10 and IL-10 biomarkers are implicated in the pathogenesis while Th17 and Th1 immune responses, via IL-17A and IFN-gamma might play protective roles.
Infection with the malaria parasite Plasmodium falciparium during pregnancy of often cause severe maternal and neonatal outcomes, which sometimes are influenced by pregnancy-induced changes in cytokine and chemokine levels. Thus, this study sought to understand the contributions of such cytokines and chemokines on the pathogenesis of pregnancy-associated malaria.
The cross-sectional study was carried out at the Marie Reine Health Centre in Etoudi, Yaoundé, Cameroon with Dr Rosette Megnekou as the head of the research group.
– I conceived, analysed and interpreted the results and drafted and reviewed the final manuscript for important intellectual content she says.
The findings from this study should provide additional clues to the development of therapeutic and preventive interventions for malaria in pregnancy.
Aire-deficient mice can be used as a model for autoimmune infertility
E. Kekäläinen, N. Pöntynen, S. Meri, T. P. Arstila and H. Jarva
In this study carried out by Finnish researchers it is confirmed that Aire-deficient mice suffer from adaptive immune system – dependent infertility. These findings indicate that Aire-knockouts can be used also as a model of male infertility of autoimmune origin.
The autoimmune regulator (AIRE) protein is a transcriptional regulator, whose most described function is to promote ectopic expression of tissue-specific antigens in the thymic medulla for presentation to developing T cells. This expression is the basis for deleting autoreactive developing thymocytes and when AIRE is missing, the autoreactive T cells are able to complete their maturation and eventually cause autoimmunity. In addition, AIRE is involved in developmental processes.
Patients that lack AIRE suffer from a disease called autoimmune polyendocrinopathy, candidiasis, and ectodermal dystrophy (APECED) that is characterized by multiple autoimmune attacks against mainly endocrine organs. In addition, infertility is a common problem in these patients.
Aire-deficient mice are used as a model for APECED and in the present study the researchers wanted to resolve whether or not the reduced fertility observed in Aire-deficient mice is dependent on the adaptive immune system, and therefore a manifestation of autoimmunity in these mice. In order to address this question lymphopenic mice without Aire, AireRag1 mice, were created
Eliisa Kekäläinen was a PhD student when this study was carried out and was involved in all of the stages of the project.
– I planned and conducted the breeding experiments, did the cell transfers, and also wrote the manuscript, she says.
Eliisa Kekäläinen says that the whole project was a spinoff of a previous project with the same AireRag1 mice that has been also published in the Scandinavian Journal of Immunology.
– We created the AireRag1 model for this other project but accidentally noted that they bred normally even though the parental Aire-deficient strain did not.
The group show that whereas Aire-deficient males had a significantly reduced breeding capacity, the AireRag1-males, who lack functional T and B cells, regained full fertility. This fertility was again lost when AireRag1-males were adoptively transferred with lymphocytes from Aire-deficient donors. Thus the Aire-deficient male mice have a lymphocyte-dependent infertility, making them a good model for human autoimmune infertility.
New insights into clinical regulatory T cell sorting
M. Bergström, A.-L. Joly, P. Seiron, S. Isringhausen, E. Modig, B. Fellström, J. Andersson and D. Berglund
In the study, by a Swedish research group, flow cytometry was used to assess peripheral blood lymphocyte profiles of young healthy individuals and patients undergoing haemodialysis treatment. The study show an overall similar immunological profile of both cohorts in spite of great differences in both age and health.
Patients that have undergone organ transplants require lifelong treatment with immunosuppressive drugs to prevent acute transplant rejection. This leads to an elevated risk for, amongst other, infections and cancer and therefore novel strategies for these patients are being investigated. One approach involves adoptive transfer of regulatory T cells (Tregs) that suppress the immune system via multiple mechanisms. To use these cells in therapy it is important to optimize and standardize the identification of Tregs and to elucidate possible differences between potential Treg donors.
In this study, the immunological profiles of healthy blood donors and patients undergoing hemodialysis treatment was analyzed. In particular, identification of Tregs using flow cytometry was performed, and the Treg-marker FOXP3 mRNA expression was studied.
– We have been able to statistically show and visually depict the differences in FOXP3 purity after flow cytometric sorting depending on which cell surface antigens and cell populations that are selected for, says Marcus Bergström, PhD student in transplantation immunology and first author of the study.
During this work, Marcus Bergström enjoyed the collaboration between various research groups and people with different areas of expertise.
– This can sometimes be challenging but ultimately very rewarding.
Marcus Bergström also appreciated the privilege of taking part in and supporting the development of cell based therapies.
– It’s a marathon, but every addition takes us a few steps closer to the to goal. We hope that our findings will come to use in the isolation of regulatory T cells for clinical therapy, for example in hematopoietic stem cell transplantation or in the prevention of kidney transplant rejection.
Collaborative effort to investigate IgE and its receptors in seasonal allergic rhinitis
M. Carlsson, L. Thorell, A. Sjölander and S. Larsson-Faria
In a study by Swedish researchers on seasonal allergic rhinitis subjects a small seasonal variation of IgE-related biomarkers studies was found, suggesting that allergen exposure affects expression of these biomarkers only to a minor degree. The objective of the study was to assess the inter- and intra-individual variability and seasonal variation of the level of IgE, high- and low-affinity receptor expression and receptor occupation in blood from seasonal allergic rhinitis subjects, both before and during a birch pollen season.
Mats Carlsson was, at the time of the study, a member of a project team developing a fully human anti-IgE monoclonal antibody, MEDI4212. The project team consisted of members in Lund and MedImmune Cambridge, UK.
– At the time I had a position as Senior Scientist at the Department of Clinical Discovery Medicine, AstraZeneca R&D Lund, Sweden, Mats Carlsson says. My main responsibility was early clinical planning and leading the biomarker program in the project. I was the main project contact for Thermo Fisher Scientific (Phadia AB), Uppsala, Sweden. Together with Anders Sjölander we lead the work on free-IgE assay development. After the completion of the study I took on the role as main author of the publication.
Mats Carlsson says that robustness for most of the assay was very good.
– As an example, the free-IgE assay was verified with results for discriminatory ability, precision, linearity and stability in comparison to the commercial total IgE assay.
Biomarkers can serve as indicators in early clinical development of efficacy and increase the confidence in the program for further development. Furthermore, robust biomarkers may be used in early clinical testing for dose selection and not at least as markers for patient stratification.
– In the perspective of developing a drug, as in this case the anti-IgE mAb MEDI4212, it is advantageous to have robust biomarkers, Mats Carlsson says.
This study was a big collaborative effort, which Mats Carlsson says was very fruitful and fun.
– First we had the collaboration with our MedImmune Cambridge colleagues to develop MEDI4212. Secondly, the work in Lund between several departments, such as Clinical Discovery Medicine, Discovery Translational Sciences and the Clinical Pharmacology Unit that took care of the subjects, was very inspiring. It is also worth mentioning Susanne Larsson-Faria and the Biomarker Team in Lund and their dedicated work with developing the flow cytometry assays and the planning of the clinical study. We also had a very fruitful collaboration with Thermo Fisher Scientific and acknowledging of the excellent work of Lisa Thorell developing the free-IgE assay on the ImmunoCap platform.
On the function of human Foxp3
R. Elhage, M. Cheraï, B. Levacher, G. Darrasse-Jeze, C. Baillou, X. Zhao, A.-M. Khatib, E. Piaggio and D. Klatzmann
Regulatory T cells (Tregs) have been shown to play a major role in the control of many autoimmune and inflammatory diseases. Yet, little is known about the molecular mechanisms that control the properties of human Tregs. In the present study, the researchers from Paris investigated the transcription factor Foxp3 that is central for the development and function of Tregs.
Human Foxp3 exists as four isoforms generated by alternative splicing. Mouse Foxp3 only exists as one isoform, but can be proteolytically cleaved by N-terminal and/or C-terminal proprotein convertase subtilisin/kexins (PCSKs).
Here, the researchers show by transcriptome analysis that the proprotein convertases PCSK7, PCSK5 and Furin are present in human CD4-positive T cells with different expression patterns.
In addition, human CD4+ cells where transduced with Foxp3-expressing lentiviral vectors and generation of proteolytically cleaved Foxp3 was detected by Western blot. Three different Foxp3 forms were detected, indicating that human Foxp3 can also be subjected to proteolytic cleavage at the N-terminal and C-terminal ends.
Thereafter, the suppressive activity associated with these three splice forms was assessed, showing that C-cleaved of N&C-cleaved Foxp3 had almost no suppressive function, indicating a crucial role of the human Foxp3 C-terminal region in the suppressive activity of T regulatory cells.
Genetically modified maize that express a bacterial protein does not exert adjuvant effects
Cry1Ab Protein from Bacillus thuringiensis and MON810 cry1Ab-transgenic Maize Exerts No Adjuvant Effect After Airway Exposure
M. Andreassen T. Bøhn, O.-G. Wikmark, J. Van den Berg, M. Løvik, T. Traavik and U. C. Nygaard
In a study by researchers in Norway and South Africa it is shown that pollen or plant material from the genetically modified maize MON810 did not promote allergic responses to a well-known unrelated allergen in a mouse model of airway allergy. This means that this genetically modified maize does not act as an adjuvant, a concern that have been raised when discussing the safety for consumers of transgenic crop plants.
The genetically modified maize used in this study has been inserted with a processed version of a transgene from the soil bacteria Bacillus thuringiensis to express proteins with insecticidal properties. Such proteins may introduce new allergens or act as adjuvants that promote allergic response. The aim of the present study was to investigate whether plant material from this transgenic maize could act as adjuvants towards the unrelated allergen ovalbumin.
A mouse model of airway allergy was used and the known pro allergenic adjuvant cholera toxin was used as a positive control. To investigate the allergic response increase in specific IgE, eosinophils and Th2 cytokines in mediastinal lymph node cell supernatants was measured.
Monica Andreassen is a PhD student in the group and was responsible for the animal experiments, that were conducted both in Norway and South Africa.
– The most fun part of this study has been to work with inspiring researchers in both countries. It was also rewarding to be involved in all the steps in the process – from the planning and the conduction of the experiments to the analyses and the scientific writing. Now I look forward to my PhD dissertation the 19th of March, she says.
The observation that the Bacillus thuringiensis protein from the transgene maize did not exhibit an adjuvant effect similar to that observed with cholera toxin means that the hypothesis of adjuvant effect from insecticidal genetically modified maize was not supported in the mouse model. However, although the doses used in these experiments may represent ”relevant doses”, long-term exposure of the protein should be included in future studies.
Antibodiy-responses against citrullinated proteins in Leishmania patients
Anti-Citrullinated Peptide Antibodies in Sudanese Patients withLeishmania donovani Infection Exhibit Reactivity not Dependent on Citrullination
E. Åhlin, A. I. Elshafie, M. A. M. Nur and J. Rönnelid
Sudanese patients infected with the parasite Leishmania donovani have antibodies against citrullinated peptides, similar to what is seen in patients with reumatoid arthritis. However, the parasite-infected patients, but not the arthritic patients, also show reactivity to the non-citrullinated form of the antigen.
This study was carried out by a Swedish group and Erik Åhlin was a PhD student at the time and did the work as a part of his thesis.
– I was responsible for the work in the lab, and I also compiled the results and analyzed the data, he says.
Infection with Leishmania donovani causes an internal disease called visceral leishmaniasis with an immunopathology characterized by a strong humoral immune response and high production of anti-Leishmania antibodies, circulating immune complexes and polyclonal activation of B-lyphocytes. As inflammation in general is associated with citrullination, the Swedish group sought to investigate antibodiy-responses against citrullinated proteins in Sudanese Leishmania patients.
– I really enjoy the field of diagnostics, it is so exciting, Erik Åhlin says. It is particularly interesting to take the test result further and to dissect what it means, both in a clinical and a methodological meaning.
The findings in the study stress the importance to interpret a positive citrullinated protein/peptide-test carefully when evaluate in non-rheumatic conditions.
Placental transfer of IgG antibodies in twin pregnancies
S. C. L. Stach, M. L. Brizot, A. W. Liao, P. Palmeira, R. P. V. Francisco, M. M. S. Carneiro-Sampaio and M. Zugaib
In a study carried out in the twin clinic of the Department of Obstetrics and Gynecology together with the Pediatric Department of Sao Paulo University it was found that IgG umbilical serum concentration against Klebsiella and Pseudomonas LPS and Group BStreptococcus is associated with specific maternal IgG concentrations and the presence of maternal diabetes.
Sonia Stach is a postgraduate student and the main researcher involved in the study, which is a part of her PhD work.
– Twin pregnancies have a high risk of prematurity and adverse neonatal morbidity, such as admission to intensive care units and infections. Neonatal immunity depends on the acquisition of intrauterine maternal antibodies. Therefore, it is important to understand factors related to IgG maternal transfer in twins to try to develop strategies to improve neonatal prognosis, she says.
Since there is no effective prevention therapies for premature delivery in twin pregnancies, it is important to make an effort to minimize the risk of newborn complications such as lung immaturity and neonatal infections. Strategies, such as mother´s vaccination may be an option in the future.
– The most exciting part of the work was to observe the results that demonstrated factors related to the transfer of immunoglobulins from mother to neonates in twin pregnancies, Sonia Stach concludes.
Characteristics of peritoneal macrophages in NOD mice
R. Emani, C. Alam, S. Pekkala, S. Zafar, M. R. Emani and A. Hänninen Rohini Emani
In the present study, researchers from Turku, Finland show that peritoneal macrophages in non-obese diabetic (NOD) mice have increased levels of activation markers soon after weaning.
The autoimmune disease type 1 diabetes (T1D) is primarily mediated by T and B cells in the NOD mouse. However, innate immune cells including macrophages also influence this disease. Macrophages play a crucial role in innate immune reactions, and peritoneal macrophages guard the sterility of this compartment. In T1D the gut microbiota and gut immune system appear to contribute to disease pathogenesis. The Finnish group have recently reported elevated free radical production and increased permeability of gut epithelium in NOD mice. This impaired barrier function could lead to bacterial leakage to the peritoneal cavity and to explore the consequences they here characterize peritoneal lavage cells from young, newly weaned NOD mice.
Rohini Emani is a PhD student in the group and the first author of the study who is currently writing her thesis.
– My main responsibilities were to study the levels of activation markers on macrophages and to analyse IRAK-M from the macrophages with Western blotting, she says.
The peritoneal macrophages displayed increased levels of IRAK-M at protein levels after weaning compared to the control strains, and became desensitized to LPS stimulation. This resulted in lower TNF-α production upon in-vitro stimulation with LPS.
In conclusion, the group demonstrate that the number of macrophages in the peritoneal cavity increases rapidly in NOD mice after weaning, ant that these cells show phenotypic characteristics of macrophages exposed to TLR ligands in vivo. In addition, they show enhanced plasma levels of LPS, suggesting translocation of gut microbes or their products into the systemic circulation.
– I enjoyed very much to discuss with my supervisor and colleague the hypothetical model of what causes macrophage influx in the peritoneal cavity and how that may connect to the pathogenesis of islet autoimmunity. The most fun was perhaps to discover that IRAK-M becomes strongly expressed in NOD macrophages after weaning and that the presence of LPS in peritoneal cavity was associated with activation of T cells in pancreatic lymph nodes, Rohini Emani says.
IL-7 – a possible new prognostic marker for acute graft-versus-host disease
T cell Reconstitution in Allogeneic Haematopoietic Stem Cell Transplantation: Prognostic Significance of Plasma Interleukin-7
K. Kielsen, K. K. Jordan, H. H. Uhlving, P. L. Pontoppidan, Z. Shamim, M. Ifversen, C. Heilmann, C. H. Nielsen, H. Sengeløv, L. P. Ryder and K. G. Müller
In this study a Danish group from Copenhagen shows that the levels of the cytokine interleukin-7 only seven days after allogeneic myeloablative haematopoietic stem cell transplantation might be able to predict the rate of T cell reconstitution and development of acute graft-versus-host disease.
Katrine Kielsen was a medical student during the work that lead to this publication, and is currently extending the findings from this work as a PhD student.
– My responsibilities were to collect and store patient samples in collaboration with three of the co-authors, to measure plasma IL-7 levels by ELISA and to perform the data analysis and draft the paper. The most enjoyable part of the work was to analyze the data in order to explain the immunological mechanisms involved in the complex reconstitution of T cell immunity, which are central for development of severe complications following allogeneic hematopoietic stem cell transplantation, she says.
Katrine Kielsen thinks that one of their main findings is the possible use of IL-7 as a prognostic marker at a time point where lymphocyte count are immeasurable. This may provide new tools for individualizing treatment
Earlier, treatment with recombinant IL-7 has been suggested for the long-lasting T cell deficiency in patients undergoing allogeneic stem cell transplantation. Here, the group finds that patients suffering from slow T cell reconstitution actually have the highest levels of IL-7, suggesting that the up-regulation of IL-7 based on less consumption by T cells might not fully compensate the lymphopenia.
– However, the association of high levels of IL-7 and alloreactivity – showed by us and others – suggest that IL-7 should be used with caution in non-T cell-depleted allogeneic haematopoietic stem cell transplantation, Katrine Kielsen concludes.
Better understanding of Mycobacterium tuberculosis infection
PstS-1, the 38-kDa Mycobacterium tuberculosis Glycoprotein, is an Adhesin, Which Binds the Macrophage Mannose Receptor and Promotes Phagocytosis
M. Esparza, B. Palomares, T. García, P. Espinosa, E. Zenteno and R. Mancilla
In this study from the January issue of the Scandinavian Journal of Immunology the authors investigate the strategies that Mycobacterium tuberculosis uses to infect host cells. Infection the critical first step in the pathogenesis of tuberculosis. Specifically, the authors investigate the pathway of macrophage infection that involves adhesins present on the bacterial surface that are recognized by receptors on the macrophage. The authors show that the microbe cell wall has several adhesins that are mainly glycoproteins.
This study is part of the PhD work of Miguel Esparza at the department of immunology at Universidad Nacional Autonoma de Mexico and was performed under the direction of professor Raul Mancilla.
After finding several adhesions on the macrophage surface, the Mexican research group chose to focus their studies on one of them, PstS-1 and shows that it is a mannose-containing glycoprotein. PstS-1 is shown to bind the mannose receptor on the macrophage, an interaction that promotes phagocytosis.
– The phagocytosis of Mycobacterium tuberculosis through the mannose receptor is of big interest because it promotes a permissive intracellular environment that favors infection, Raul Mancilla says.
Raul Mancilla enjoyed performing this study, and especially he liked the close interaction with his students during the research. He also believes that their findings can be important for future drug development.
– Mycobacterial adhesins are potential targets for vaccine development!
The redoxome – a complicated system that can react in unexpected ways
Bøyum, R. J. Forstrøm, I. Sefland, K. L. Sand andH. B. Benestad
In this publication, the authors examine if a simple, in vitro chemiluminiscence set-up with redox components from human polymorphonuclerar neutrophils (PMN) and red blood cells (RBC) can clarify some unexplained workings of the redoxome. The authors present proof-of-principle evidence that the assay can assess redox effects, and demonstrate the intricacies of redox reactions.
The redox potential is determined by an interacting system of redox enzymes and sulfhydryl proteins, as well as their substrates: smaller molecules such as NADPH/NADP and superoxide – they all together constitute the redoxome.
The redoxome can be said to ”buffer” a cell´s electrochemical (redox) potential similarly to how buffering of pH is important for the function of enzymes and other cellular constituents.
Arne Bøyum, MD, is a senior investigator, 86 years old, and the first author of the paper. He designed the experiments and did most of the laboratory work.
– It was a pleasure, at my age, to find unexpected, or unforeseeable responses that might be of value in further work, he says.
One of the surprising findings was that calproctin and its component peptides A8 and A9 make up a surprisingly high concentration in PMN. Several functions have been ascribed to these proteins but rarely redox functions.
After a series of various dose-response experiments, utilizing their chemiluminescence assays and different reactants, the authors suggest that the redoxome is a complicated system that can react in unexpected ways. This makes it very difficult to foresee the results of perturbations in the intact organism and they describe prescriptions of antioxidants to an intact organism as ”risky business”.
On chemokine-levels in children with type 1 diabetes
S. U. Thorsen, S. Eising, H. B. Mortensen, K. Skogstrand, F. Pociot, J. Johannesen, J. Svensson and on behalf of the Danish Childhood Diabetes Registry
In a study performed by researchers at Copenhagen University levels of cytokines is found to differ according to age, time period and season among children newly diagnosed with type 1 diabetes (T1D) and their healthy siblings. T1D is a T cell-driven autoimmune disease caused by a lack of tolerance towards beta cells in the pancreas. Auto-reactive T cells migrate to the islets of Langerhans where they destroy the beta cells. However the mechanism by which these T-cells migrate to the islets is not very well known, but studies have shown a role of chemokines in the pathogenesis of T1D. Therefore, the aim of this study was to elucidate the role of inflammatory chemokines in T1D at time of diagnosis. The hypothesis was that systemic levels of inflammatory chemokines in patients with recent onset T1D is different from their health siblings and that gender, age, time period, season and autoantibody status influences the chemokine levels.
– However, we found very few differences in levels of the chosen chemokines between cases and controls, says Steffen Thorsen, first author of the paper. But we saw that levels of chemokines vary with season, period and age.
Steffen Thorsen was responsible for the analysis and the interpretation of the data in the article. The data was derived from a population-based register of children with T1D, the Danish Diabetes Register. It contains information on more than 3500 newly diagnosed patients aged 0-15 years with an associated biobank with blood samples from the children and their first-degree relatives.
From this study it becomes clear that it is important to match or adjust for especially season and period when conducting new studies concerning chemokine levels in children and adolescents.
New insights into human peripherally induced regulatory CD8+T cells
U. Bjarnadottir, A. L. Lemarquis, S. Halldorsdottir, J. Freysdottir and B. R. Ludviksson
In a study from an Icelandic research group it is delineated how the innate immune system is capable of directly influence both the de novo development and function of human peripherally induced regulatory CD8+ T cells (CD8+ iTregs).
– In particular we demonstrate that the initial pro-inflammatory response has a negative tolerogenic effect mediated through reduced IL-10 and TGF-beta1 driven mechanism, says Björn Ludviksson, principal investigator.
The study provides direct evidence for the role of IL-2 and TGF-beta in human CD8+iTreg differentiation and that their suppressive function is inhibited in the presence of IL-1beta and TNFalpha, which may be linked to their effect to reduce IL-10 and TGF-beta1 secretion.
– One of the greatest joy of all scientific work is to participate in a collaborative effort involving lot of hard working people dedicated to tackle difficult basic biological questions. The most fun regarding this work in particular was to discover how significant proportion of the CD8+ human T-cells can be induced into development of iTregs and the major regulatory role of the innate immune system upon that pathway, Björn Ludviksson conlcudes.
D3G – a candidate gene for autoimmunity?
B. Gokturk, S. Keles, M. Kirac, H. Artac, H. Tokgoz, S. N. Guner, U. Caliskan, Z. Caliskaner, M. van der Burg, J. van Dongen, N. V. Morgan and I. Reisli
CD3G, the gene for the CD3gamma chain, part of the T-cell co-receptor, should be studied as a candidate gene for autoimmunity, suggests a group from Konya Training and Research Hospital in Turkey. In a study in the November issue of Scandinavian Journal of Immunology the group reports frequent and multiple autoimmune findings in patients with CD3gamma deficiency. In the study, three new CD3gamma-deficient siblings from a consanguineous family is reported. Further a previously reported non-consanguineous family with two CD3gamma-deficient siblings with the same mutation was re-evaluated. All five patients displayed several different autoimmune symptoms, but also family members that are heterozygous carriers of mutations display autoimmunity.
Bahar Göktürk who directed the research in this study, says that CD3G could be a candidate gene for autoimmunity.
– I think CD3 gamma deficiency is more than we expected especially in countries that have high frequencies of consanguinity. I hope that this study can create an awareness about the association between autoimmune diseases and immune deficiency.
CD3gamma-deficiency is a very rare condition, up to date only nine patients have been described, including the ones in this study. In addition it seems like siblings with the same mutation may have different clinical courses, which may be due to existence of undefined modifying genes in these individuals. Interestingly, CD3gamma-deficient patients have normal immunoglobulin levels and antibody responses, and not even T cell counts or functions might be affected. In the present study, only one of the patients had markedly abnormal T cell functions.
In conclusion, the authors suggest that CD3gamma-deficiency should be kept in mind when investigating patients with autoimmunity and atopic diseases
Allergy to flowers, a potential problem for florists
Albert van Toorenenbergen
In the present report by retired clinical chemist Albert van Toorenenbergen the IgE response towards decorative flowers was studied. The author describes the pattern of IgE binding to allergens in freesia, gerbera and chrysanthemum pollen. It is concluded that occupational exposure to many different flowers can induce IgE against these flowers.
– Before my retirement as a clinical chemist I supervised the routine analysis of specific IgE antibodies. Clinicians in the Department of Allergy occasionally asked for allergen-specific IgE tests, that were not commercially available, Albert van Toorenenbergen, says.
For the purpose, a CNBr-activated Sepharose bead procedure for preparation of solid-phase allergen extracts was used. With this procedure tests for IgE against pollen from some decorative flowers, sweet bell pepper pollen, and more, were set up.
– Shortly before my retirement I realized that I had not yet published such experiments for pollen from some decorative flowers, performed earlier, and decided to write the present paper.
In the paper, sera were obtained from patients with symptoms associated with their occupational handling of flowers; they worked in greenhouses or florist shops, and one was a student at a florist school. With the limited number of sera tested, cross-reactivity between mugwort pollen and pollen from freesia, gerbera and chrysanthemum pollen was found.
– I find the demonstration of IgE against rare allergens an exciting aspect of work like this, Albert van Toorenenbergen says.
The diagnosis of an IgE-mediated allergy to decorative flowers drove one of the patients to drop out of florist school and one to stop working in a florist shop.
Gut microbiota of the mother is important for the immunological status of the offspring
N. Tormo-Badia, Å. Håkansson, K. Vasudevan, G. Molin , S. Ahrné and C.M. Cilio
Alteration of the maternal gut microbiota during pregnancy permanently alter the gut microbiota in the offspring, is shown by Neivis Tormo Badia and co-workers in a study published in the October Issue of the Scandinavian Journal of Immunology. This seems to be important for the establishment of tolerance, ultimately modulating the development of diabetes in the offspring.
In an earlier human prospective study, it was shown that children with type 1 diabetes had a decreased percentage of Firmicutes, whereas species belonging to Bacteriodeteswas increased in the same children. Because of this link between the gut microbiota and type 1 diabetes, the authors wanted to investigate the effect of antibiotic treatment during pregnancy. To this end, female NOD mice was treated during gestation with a mix of broad spectrum antibiotics. The offspring was thereafter examined for immunological status, alterations in the gut microbiota and diabetes development.
Neivis Tormo Badia was a PhD student in the group at Lund University and did the practical work with the animal model as well as analysis and interpretation of the results.
– Because of the clinical relevance in our hypothesis it was very exciting to analyse the results of the study and to find out whether the data fits with our hypothesis that antibiotics treatment during pregnancy would affect the outcome of diabetes development, she says.
New insights into the expression pattern of T Cell-specific adapter protein in healthy human tissues
A. D. Pandya, T. B. Leergaard, E. Dissen, G. Haraldsen and A. Spurkland
The current study, published in the September issue of the Scandinavian journal of immunology, is the first to address to what extent T cell-specific adapter protein (TSAd) is expressed in various tissues in situ, including lymphoid and non-lymphoid tissue.
TSAd is expressed in activated T cells, NK cells and endothelial cells, but due to previous lack of suitable reagents, its tissue expression has not been mapped before. The protein plays a role in cell signalling, proliferation and cellular migration, but its precise role in these processes is yet not well established.
The main findings from the current study include:
I) In lymphoid tissues, a fraction of CD3 expressing T cells also expressed TSAd.
–This may indicate that a particular T cell subset expresses TSAd. It will be more interesting to quest about this subset in future, says Abhilash D. Pandya, PhD student in the group of Anne Spurkland
II) In Mucosa-associate lymphoid tissues (MALT), the group observed that all the intraepithelial lymphocytes (IELs, specifically T cells) expressed TSAd.
III) In non-lymphoid tissues, they observed TSAd expression in Langerhans’ islets of pancreas. However, the identity of TSAd expressing islet cells, and the significance of TSAd expression in these cells is yet to be established.
– We did not detect TSAd expression in the endothelium of non-lymphoid organs including liver, kidney, lung and heart as well as in lymphoid tissues, Abhilash D. Pandya says. But we observed TSAd expression in endothelium of blood vessels in skin. This indicates varied expression of TSAd in endothelial cells, based on endothelium of specific tissue. Henceforth, it could be that the TSAd level in some endothelium is normally below the limit of IHC detection.
Abhilash D. Pandya developed the tissue specific IHC protocol for the study, and also performed a series of experiments including analysis. He also wrote the manuscript.
– I really enjoyed working on such a novel project, especially to analyse and conclude the results!
One step closer to solving the riddle of early pregnancy termination.
N. Sotnikova, D. Voronin, Y. Antsiferova and E. Bukina
In this study Natalia Sotnikova and co-workers set out to detail the mechanisms of the interaction of decidual CD56+ NK cells (dNK), infiltrating the maternal part of placenta, and trophoblast cells of foetal origin. They conclude that unbalanced activation of dNK can lead to the impairment of dNK and trophoblast cells interaction during RSA.
There is a unique immunological situation in pregnancy when a semiallogenic foetus is implanted and successfully developed within a mother´s womb. This is called the immunological paradox. We do not know exactly why the mother´s immune system does not reject the foetus. Following implantation, the trophoblast cells, which are the primary foetal cells, invade the decidual tissue of placenta of maternal origin and for the pool of so-called extravillous cytotrophoblast cells, which participate in the development of early placenta. At this stage of the pregnancy and unique population of CD56bright NK cells has been shown, but the biological role of these cells has not been demonstrated.
The present study was designed to detail the mechanisms of the interaction of dNK and trophoblasts during normal and pathological early pregnancy (first semester recurrent spontaneous abortion, RSA) and to concretize the role of dNK in trophoblast behaviour.
– The main finding of our work is proving the cross-talk between decidual CD56+ NK cells and trophoblast in normal and RSA pregnancy, Natalia Sotnikova, head of the laboratory of Clinical immunology at the Federal State Research Institute of Maternity and Childhood in Russia says.
From one side the maternal immune system actively recognize foetal antigens and respond to these antigens, but normally this response does not lead to foetus damage. When this physiological process is impaired either due to inadequate maternal response or due to inadequate foetal antigen stimulation the immune balance between mother and foetus is crashed and foetus is rejected.
Natalia Sotnikova was the supervisor of the research and she planned the research, participated in the selection of the patients, discussed experiments with co-workers and wrote the manuscript. The work in the lab is not always calm.
– We had some rivalry with our colleagues from the Laboratory of Morphology over clinical samples, like who would have the next piece of placenta, but in the end everything was well and everyone got the samples they needed.
Natalia Sotnikova is very pleased over the results and the fact that we are now one step closer to solving the riddle of early pregnancy termination.
– The fine immune mechanisms participating in this process are very far from complete understanding, but we can confirm that the dramatic stimulation of the functional activity of the maternal decidual CD56+ NK cells by foetal trophoblast cells is one of the mechanisms behind early pregnancy termination.
New epitopes from essential tumor antigens – a step toward more efficient cancer vaccines
H. L. Larsen, M. H. Andersen, H. H. Wandall, C. B. Madsen, R. E. Christensen, T. R. Petersen and A. E. Pedersen
In a study by Hjalte Larsen and coworkers, presented in the August issue of Scandinavian Journal of Immunology the group show generation of a robust CD8+ T-cell response against an endogenous target protein, Bcl-XL. Delivering in silico predicted MHC-I antigens in an immunogenic setting by display on in vitro differentiated dendritic cells elicited a specific T-cell response by breaking immunological tolerance. The successful induction of an immune response targeting an endogenous tumor-associated antigen might facilitate the development of more reliable preclinical models of cancer immunotherapy.
Hjalte Larsen was a master student in the group of Dr AE Pedersen at the University of Copenhagen at the time of the study. The project was part of his master thesis and he performed the majority of the experimental work and data analysis as well as designed the experimental setup.
– The most fun part of the work was the successful transition from in silico prediction of target epitopes into the production of a cell-based vaccination strategy that succeeded in inducing specific immunological responses.
Active immunity against tumor-associated antigens to prevent relapse of cancer is a promising, but so far not very efficient therapy, possibly due to clonal escape of tumor cell variants. Tumor-associated antigens are essential for continued tumor cell survival and identification of these will help improve preclinical development of cancer immunotherapy.
Bcl-xL is an anti-apoptotic protein that have been suggested to be an essential tumor antigen. Lack of well-defined murine epitopes have made targeting for the use in cancer vaccines difficult.
Here, the group report the identification of two novel murine tumor-associated epitopes from Bcl-xL.
A recent success of a dendritic cell-like vaccine against metastatic prostate cancer is believed to lead to breakthroughs in vaccination for other types of cancers. However, to develop new cancer vaccines and immunotherapies, preclinical testing in mouse models is essential. In these models the use of endogenous tumor antigens is used, but so far relatively few such antigens have been characterized and tested.
Novel therapeutic approach for auto-inflammatory diseases
I. Nikolic, M. Vujicic, I. Stojanovic, S. Stosic-Grujicic and T. Saksida
Here, the group from University of Belgrade shows that carbon monoxide has an anti-inflammatory effect through the regulation of the balance between pro-inflammatory Th1/Th17 and anti-inflammatory Th2 cells.
Ivana Nikolic, the first author of the paper is a PhD student at her final year of studying.
– My main responsibility in the work that led to this paper was to perform the experiments in collaboration with other co-authors, to draw the figures and the draft version of the manuscript.
Carbon monoxide is endogenously produced by haeme oxygenase-1 and as profound effects on intracellular signalling processes, generating anti-inflammatory, anti-proliferative and anti-apoptotic effects. CORM-A1 is a boron-containing compound that can release carbon monoxide is such a way to mimic physiological functions of haeme oxygenase-1. In the present study the group studied the immune-modulatory effects of CORM-A1 on murine lymph node-derived T cells in vitro and its influence on T-cell proliferation, activation and differentiation.
– In my opinion, Ivana Nikolic says, the main findings of the paper is that carbon monoxide releasing molecule CORM-A1 affects the Th cells differentiation in vitro, and therefore I feel this study offer novel therapeutic approach in managing Th1/Th17-mediated auto-inflammatory diseases.
Novel lectin-lectin interaction – a crucial step in homing and lung entrapment of mesenchymal stromal cells
H. Suila, T. Hirvonen, A. Kotovuori, I. Ritamo, E. Kerkelä, H. Anderson, S. Natunen, J. Tuimala, S. Laitinen, J. Nystedt, J. Räbinä andL. Valmu
In the publication by Heli Suila and co-workers in the July issue of Scandinavian Journal of Immunology a novel ligand for P-selectin is described; galectin-1. This interaction was seen on human multipotent mesenchymal stromal cells, a cell type that have the ability to differentiate into various cellular lineages and can be expanded in culture conditions without losing their multipotency. These cells can modulate immunological responses and home to sites of ischemia or injury and have been therapeutically explored in cell-based therapy in several different diseases such as multiple sclerosis and type 1 diabetes. However, a major problem in cell therapy is the inefficient homing of transplanted cells to desired organs or tissues. This homing of stem cells seem to use similar mechanisms as leukocytes do, and one of the important molecules in this process is the lectin P-selectin. This molecule has glycogonjugate ligands, the main one is PSGL-1. However P-selectin has previously been shown to bind to mesenchymal stromal cells, even though they do not express this ligand. Therefore the group from Finland set out to find the P-selectin ligand on these cells, and surprisingly found another lectin: galectin-1.
Heli Suila was a PhD student at the Finnish Red Cross Blood Service when the study was undertaken. She was responsible for most of the lab work as well as for writing and putting the paper together. The collaboration between the co-authors in the lab was very rewarding and fun, she says.
– We had lots of good laughs and fruitful discussions while working long days with this project.
This is the first interaction that has been described between two lectins, and it may play a role in the immunomodulatory targeting of umbilical cord blood derived mesenchymal stromal cells. Further, the interaction between P-selectin and galectin-1 may play a role in the undesired lung entrapment of intravenously infused mesenchymal stromal cells that occur when trying to use these cells in therapy.
Study with obstacles on Interferon lambda-2 in tbc patients
M. Travar, M. Vucic and M. Petkovic
A group of researchers from Banja Luka University in Bosnia and Herzegovina has shown that inflammatory cells in sputum from tuberculosis patients can be a source of interferon lambda after receptor stimulation by Mycobacterium tuberculosis components. Interferon lambda is a new (discovered in 2003) group of cytokines, classified as type III interferon.
– The role of interferon lambda in pulmonary tuberculosis control was the topic of my doctoral dissertation at the Faculty of Medicine at Banja Luka University, Maja Travar, the first author of the paper, says.
– I thought that the role of newly discovered interferons in tuberculosis infection could be interesting to do, since the type III interferons are mostly connected with the control of viral infections, and there were only few publications about their role in bacterial infections control, and none of IFN lambda level in tbc, she explains.
The study was conducted to measure interferon lambda-2 levels at the site of infection which could be helpful to determine if immune stimulation by Mycobacterium tuberculosis in lung occurs.
– It is too early to conclude that the findings of this study can tell whether infection is present or not, firstly due to small sample size (we obtained adequate samples from only 51 patient with active infection, and only 15 newly discovered) and the presence of few outliners in all study groups. However, interferon lambda-2 measurement in combination with clinical findings could be more helpful.
The group from Baja Luka had a lot of obstacles to overcome during this study. Bosnia and Herzegovina is a very poor country and finding the financial resources for research is very difficult. The group really wanted to measure mRNA from the samples along with ELISA, but they simply had no financial for that. In addition, Maja Travar also had other things to tend to during the study.
– When we conducted the ELISA, I was taking care of my baby during pauses (she was only few months old). It is very challenging to be a mother and a researcher. And I have to say that this paper was written mostly during nights (since I have three small children), but during this research I was full of energy and I really enjoined it.
Expression of costimulatory factors on dendritic cells to be used in anti-tumour vaccine
High functional CD70 expression on alpha-type-1-polarized Dendritic cells from patients with chronic lymphocytic leukaemia
K. Junevik, O. Werlenius, L. Fogelstrand, A. Karlsson-Parra, P.-O. Andersson
In a study performed in the PO Andersson lab at Sahlgrenska University Hospital in Gothenburg, means to optimize the method of using antigen-loaded dendritic cells (DCs) as anticancer vaccine, was investigated. A DC vaccine should elicit an effective T cell anti-tumour response. To do this, a DC needs to provide an array of functions, such as antigen-presentation, costimulation, induction of effector functions and expression of receptor with tumour-specific homing properties.
The most frequently described DC vaccine today is being matured with a cocktail containing prostaglandin E2, and they are called PGE2DCs. However, these cells tend to give an insufficient Th1 response. Instead, it has been suggested that alpha-type-1 polarized DCs (called aDC1) give a more appropriate activation of effector cells. It has also been shown that the costimulatory factor CD70 appears to be essential for optimal DC function. So far PGE2 is suggested as one of the most important factors for the induction of CD70 on DCs. The present investigation shows that aDC1s express CD70 equally good as PGE2DCs.
Katarina Junevik was a PhD student in the group when the study was conducted. She defended her thesis in September 2013, and she did almost all the laboratory work. The study design and writing of the paper was done together with her supervisor PO Andersson
– The most fun part was to create a method and make it work, like we did. To be able to with so few cells still create a milieu where we could analyse the production of cytokines was fascinating, she says.
Expression of costimulatory surface receptors after dendritic cell maturation appears to be a dynamic process. To establish an optimal anti-tumour DC vaccine protocol, proper timing for the determination of such receptors are crucial.
Removal of IFNg and perforin tips the balancing race between virus and host in favour of the virus
IFNγ and perforin cooperate to control infection and prevent fatal pathology during persistent gammaherpesvirus infection in mice
C. Bartholdy, M. Høgh-Petersen, P. Storm, P. J. Holst, C. Ørskov, J. P. Christensen and A. R. Thomsen
The cooperation of IFN-gamma and perforin to control an viral infection and prevent immune-mediated as well as virus-mediated pathology is shown in this study by Christina Bartholdy and co-workers.
Christina Bartholdy was a PostDoc in the group at the time of the study, and now works as a senior scientist at Novo Nordisk A/S. I ask her about her role in the study.
– I was the main driver of the research process including planning and conducting the majority of the experiments, interpreting the results, designing follow up experiments and writing the paper. However, the whole process that led to this publication has been a close and fruitful collaboration with especially professor Allan Randrup Thomsen, but also with significant scientific contributions from the other authors.
Inspired by a paper by Dutia et al. showing that MHV68 infection of IFNgR-/- mice lead to splenic atrophy and fibrosis, and that this pathology could be reversed by depletion of CD8 T cells, the initial research question was to analyse whether this seemingly pathological role of CD8 T cells was mediated by perforin-mediated killing of virus infected cells. Much to the researchers´ surprise, they observed that although absence of perforin effectively prevents splenic atrophy, it likewise results in an inability to control the virus infection, which leads to chronic immune activation, significant immunopathology and finally complete destruction of the normal architecture of the spleen. In other words, removal of both IFNγ and perforin seems to tip the balancing race between the virus and the host in favour of the virus, which subsequently results in fatal pathology most likely due to chronic immune activation.
What was most fun during the work that led to this paper?
– One exciting and challenging thing about biological research is that the pathway from scientific hypothesis to conclusion is rarely as straight forward as you initially think. This has indeed also been true for this project. Interpreting our surprising findings and designing new experiments to further address the biology behind has been really fun and exciting, Christina Bartholdy concludes.
Gut microbiota and blood acute-phase proteins
W. Schrödl, B. Kleessen, L. Jäkel, A. A. Shehata and M. Krüger
In this study from the University of Leipzig it is shown that some special bacterial groups of the gastrointestinal tract microbiota have an influence on the concentration of special acute-phase proteins in the blood of cattle.
–This is an indication that special groups of bacteria in the gastrointestinal tract can induce inflammation reactions, says Wieland Schrödl, Veterinary and scientist at the University of Leipzig employed in the Institute of bacteriology and mycology.
A second new finding in this study was the negative acute phase reaction of the C-reactive protein in cattle. In human, dog and pig it is a positive acute phase reactant.
– I was very happy to be able to measure a clear positive correlation between the acute phase protein LBP in blood and the quantity of aerobic gram-negative bacteria and clostridium perfringens in the gastrointestinal tract of cows. On the other side, I have determined a negative correlation between the number of lactobacilli in the gastrointestinal tract and the positive acute phase proteins LBP and haptoglobin. The importance of lactobacilli in the microbiota has been known, but I was able to measure it in this complex system of microbiota and immune system, Wieland Schrödl concludes.
Optimized protocol for generation of specific immunosuppressive human monocyte-derived macrophages
S. Mia, A. Warnecke, X.-M. Zhang and R. A. Harris
In this paper from the Robert Harris group at Karolinska Institutet a panel of polarization protocols for blood derived human monocytes was investigated and compared. The study was undertaken to address how best to induce in vitro an immunosuppressive human macrophage phenotype.
Macrophages are transformed from circulating monocytes into either pro-inflammatory M1 macrophages or immunosuppressive M2 macrophages. The relative balance of M1 cells driving inflammation, and M2 cells regulating the immune response, will determine the final outcome of either chronic inflammation or healing. Manipulation of this M1/M2 balance through adoptive transfer of in vitro pre-activated macrophages is thus a highly interesting therapy. An important aspect will be the stability of the induced anti-inflammatory phenotype.
During the studies of protocols to generate M2 macrophages the group made three major findings:
(a) that a stable, efficiently immunosuppressive M2 phenotype can be induced by specific cytokine stimulation in vitro.
(b) that pre-differentiation with M-CSF prior to M2 induction yields the most efficient final phenotype. This defines the final protocol suggested for use in the clinic.
(c) demonstration that it is possible to induce the immunosuppressive M2 phenotype in monocytes recovered from patients with pro-inflammatory diseases. This indicates proof-of-concept for further use in clinical immunotherapy settings.
Muhammad Sohel Mia, MD, PhD was a PhD student in the group and did recently graduate. He performed the majority of the work included in the paper, and conducted similar phenotypic analyses of mouse macrophages that led to this study of human monocytes.
– When I started working with human monocytes I expected it would be a straightforward approach as I have been working with rodent macrophages before. However, it became obvious at an early stage that human cells do not behave in a similar way to murine cells, and variability between donors was much more extreme than between individual animals. There are number of different methods used in purification of human monocytes and comparing and contrasting the different isolation and purification methods and their effects on human monocytes activation was the most fun part of the project and what lead to this paper.
Sulfatide does not modulate the course of type 1 diabetes in NOD mice
S. Rhost, L. Löfbom, J.-E. Månsson, A. Lehuen, M. Blomqvist and S. L. Cardell
In this paper from Susanna Cardell´s lab in Gothenburg it is shown that the course of type 1 diabetes (T1D) in non-obese diabetic (NOD) mice cannot be modulated by the glycosphingolipid sulfatide, which is something that has been published earlier (Lakshmimathy Subramanian et al, PLOS ONE 2012).
The study was initiated to investigate immune reactivity to sulfatide in NOD mice and the ability of the sulfatide treatment to modulate T1D development. It has been shown previously that human T1D patients, but not patients with type 2 diabetes nor healthy individuals have autoantibodies against sulfatide in serum, suggesting that sulfatide induces an immune response in the natural course of T1D in humans. In the present study, sulfatide was investigated as an autoantigen and a modulator of autoimmune disease in NOD mice. The results show that some NOD mice develop autoantibodies towards sulfatide with age, however it did not correlate with T1D progression. These data are thus in contrast to what has been described for human patients with T1D.
– This suggests that the murine model for T1D, the NOD mouse model may possess differences in the immune response towards sulfatide compare to humans in relation to the T1D development, says Sara Rhost, first author of the study.
Sara Rhost is a former PhD student in the research group of Susanna Cardell. She is now a postdoc at Monash University in Melbourne, Australia, and she thinks that working together with highly motivated and skilled collaborators was the most rewarding during the work that led to this paper.
Impaired classical complement activity in extreme preterm neonates
A. S. Grumach, M. E. Ceccon, R. Rutz, A. Fertig and M. Kirschfink
In a study by Anete Grumach and co-workers the relative immaturity of the complement system and its regulation in premature infants is demonstrated.
The proteins of the complement system are synthesized early in foetal life, although with a relative deficiency of most of the complement proteins in comparison with adult levels.
This study was not restricted to evaluate a specific population but evaluates protein regulators of the complement system not studied by others before.
Complement activity, levels of mannan-binding lectin, complement regulators and C3a as a marker of complement activation were assessed in three groups of healthy new-borns:
1) prematures (≤ 34 weeks)
2) late prematures (>34 – <37 weeks)
3) term neonates (≥ 37 weeks)
It was found that functional impairment of the classical pathway activity existed in almost all samples, with higher levels in term babies. Cord blood levels of quantitative and functional C1 inhibitor and of factors H and I progressively increased with gestational age.
– The findings may reflect not only the immaturity of complement system but it could suggest an additional factor predisposing to infections, says Anete Grumach who, together with Dr Michael Kirschfink, was responsible for planning the project.
The results showed that extreme preterm neonates lack total classical complement activity. Although the low titres of classical and alternative pathway functions are considered as risk factors for infections, an impaired regulation may pose an additional risk for uncontrolled complement-mediated inflammatory tissue destruction.
– The sample collection was extremely hard considering the gestational age, says Anete Grumach.
For me as a paediatrician, the evaluation of the results was very interesting and also the comparison with other studies.
New prognostic marker in stem cell transplantation
K. K. I. Jordan, I. J. Christensen, C. Heilmann, H. Sengeløv and K. G. Müller
In a study performed by the group of K. G. Müller, it was found that measurements of C-reactive protein (CRP) in patients that undergo allogeneic stem cell transplantation can help predict the survival after the transplant. Before the transplant, patients undergo chemotherapy and total body irradiation. This is called pretransplant conditioning and prevents graft rejection and reduces the number of remaining leukemia cells.
Karina Jordan, MD and PhD student is the first author of the paper.
– I find it important to evaluate the level of CRP before the initiation of the conditioning as the prognosis on survival is largely impaired with elevated CRP levels, even at baseline before the start of the conditioning. Prospective studies will show whether it is possible to improve survival by including CRP in the pre-transplant risk stratification profiling of these patients
The present study was a population-based retrospective study of 349 patients undergoing allogeneic stem cell transplantation at the National Danish SCT centre between January 2000 and January 2009. This is one of the first study that address the prognostic value of baseline CRP levels before start of transplantation and it was found that elevated pretransplant CRP levels predict a poorer survival.
Karina Jordan, who wrote this paper as a part of her PhD was responsible for all the data-management and did the analysis together with a statistician. She enjoys the thrill of science:
– It is always interesting to explore new areas in science, and although it is a small contribution in the big sea I hope the findings will add up to improve the morbidity and mortality of this treatment.
New insights into the pathogenesis of multiple sclerosis
M. Kostic, T. Dzopalic, S. Zivanovic, N. Zivkovic, A. Cvetanovic, I. Stojanovic, S. Vojinovic, G. Marjanovic, V. Savic and M. Colic
The relationship between Th17-mediated inflammatory and excitotoxic events was investigated during an active phase of multiple sclerosis (MS). The main pathological feature of MS is immunoinflammatory-mediated demyelination. This demyelination is often accompanied by neurodegeneration, mainly in the form of axonal degeneration, which could be initiated by glutamate excitotoxicity.
– The starting idea for this paper originated from PhD thesis, and in cooperation with my senior mentors and clinicians, we made the study design, says Milos Kostic.
Dr Milos Kostic is assistant at the Department of Immunology at Medical Faculty, University of Nis, Serbia, were this work was mainly carried out. The work was done under the ongoing project ”Preventive, therapeutic, and ethical approach in preclinical and clinical studies of genes and redox cell signalling modulators in immune, inflammatory and proliferative cell response.” supported by The Ministry of Education and Science of the Republic of Serbia.
– I would highlight, as our main finding that we have reported direct positive correlation between IL-17A and glutamate levels in cerebrospinal fluid of MS patients. This potentially suggests that Th-17 cells besides involvement in immunoinflammatory-mediated demyelinization might use glutamate excitotoxicity as effector mechanism, and thus also mediate neurodegenerative processes during MS development.
An unexpected finding in the study was that CSF IL-17A level tends to fall with disease duration indicating that Th-17 cells are possibly more important for disease initiation rather than progression which may be mediated by some other, inflammation-independent mechanisms.
How tumours evade the immune system
R. P. A. Wallin, V. S. Sundquist, E. Bråkenhielm, Y. Cao, H.-G. Ljunggren & A. Grandien
We all know that cytotoxic cells kill their target cells. But in this paper the authors conceptually show that cytotoxic cells also have an important role in fighting tumours and possibly also infections by influencing the tissue environment.
Reports have been published that tumours are selected to resist T-cell immune attacks by a variety of mechanisms including recognition and cytotoxic killing.
Proteins of the anti-apoptotic BCL-2 family such as BCL-XL have long been seen to be upregulated in many types of tumours. These proteins block the mitochondrial pathway of apoptosis and it is believed that BCL-XL protects the tumor cells from various cell death stimuli in the harsh environment of the growing tumour.
Started 15 years ago
Robert Wallin, the first author of that study, reveals that the work started almost 15 years ago, when he was a PhD student, together with Valentina Screpanti.
– We continued the work as post docs and eventually, almost 15 years after the start, the study is now finished, Robert Wallin tells.
He was until recently a group leader and associate professor at the Karolinska Institutet, but is now responsible for education development at a new life science institute in Jakarta, Indonesia, International Institute for Life-Sciences.
Robert Wallin and Valentina Screpanti worked as PhD students together in a project concerning the role of apoptosis in T and NK cell killing of infected cells and tumour cells. They found that BCL-XL protects a tumour, in vivo, from NK cell activity, but did not understand the mechanism, more than that it was not protection against the cytotoxicity of the NK cells. Valentina later found that the activity of NK cells inhibit blood-vessel expansion in the tumour in an IFN-g dependent way and that BCL-XL enables the tumour to grow in spite of the lower oxygen and nutrient supply.
Lymphocytes are important in tissue remodelling
– One interesting aspect of publishing this study is that we can draw the attention to this potentially important field, says Robert Wallin.
A few articles were published over ten years ago about the fact that another immune cell could inhibit vessel expansion in a similar way, but not much has happened since.
– I believe that the role of lymphocytes in tissue remodelling in inflammation and in tumours is an interesting field to study, Robert Wallin concludes.
On the relationship between influenza and type 1 diabetes
M. Svensson, A. Ramelius, A.-L. Nilsson, A. J. Delli, H. Elding Larsson, A. Carlsson, G. Forsander, S. A. Ivarsson, J. Ludvigsson, I. Kockum, C. Marcus, U. Samuelsson, E. Örtqvist and Å. Lernmark, the Better Diabetes Diagnosis (BDD) study group
Different viruses have been implicated in the triggering and promoting of islet autoimmunity and progress to the onset of clinical disease, but the exact role they play is still uncertain. In light of the reports on increased incidence of narcolepsy after the Swedish vaccination campaign against A(H1N1)pdm09 2009-2010, the researchers in this study wanted to investigate whether the vaccination may had altered any characteristics of type 1 diabetes in newly diagnosed children. They also determined the levels of A/H1N1-HAAb in a standard radio binding assay, a method commonly used to detect islet autoantibodies.
Type 1 diabetes is a complex disease
The etiology to type 1 diabetes is complex, and includes both genetic and environmental factors. At present, type 1 diabetes is viewed as a two-step disease. The first step is the triggering of islet autoimmunity in the genetically at-risk (HLA-DQ8, HLA-DQ2 or both), which is reflected by the appearance of islet autoantibodies. The second step is the developing of clinical disease, which may require many years to manifest. However, it is known that a higher number of autoantibodies predicts the disease progress, so that patients with multiple autoantibodies progress faster to clinical onset.
Matilda Svensson is a medical student who joined the research team in 2012 when she was working on a project as part of a summer course at Lund University. This summer project is what led to the publication.
– The study design was initiated before I joined the project, Matilda Svensson tells me. All raw data had been collected, and my task was to, together with Ahmed Delli, compile it and perform the necessary statistical analysis.
– We found that the youngest children (<3 years of age) had significantly lower frequency of A/H1N1-HAAb than older children (divided into age groups 3<6, 6<9, 9<13 and 13-18 years), which probably cannot only be explained by a lower vaccination coverage in this age group.
Vaccination may accelerate or delay progress into type 1 diabetes
The group also found that children carrying HLA-DQ2 (not together with HLA-DQ8) may have had lower frequencies of A/H1N1-HAAb, which is a new finding. Nothing was found that pointed towards an increased number of patients diagnosed with type 1 diabetes the winter season of 2009-2010 compared to seasons before and after. However, when comparing children diagnosed before, during and after the vaccination campaign, it was found that frequencies and levels of two islet autoantibodies (GADA and ZnT8QA) were increased among patients diagnosed during (GADA and ZnT8QA) and after (ZnT8QA) compared to them diagnosed before the vaccination campaign. Also, less children than expected in the youngest age group (<3 years of age) carrying the highest genetic risk (HLA-DQ2/8), where diagnosed after the vaccination campaign. These findings allowed the group to speculate that the vaccination (or the virus itself) may have accelerated, or delayed, progression into clinical disease in different children, depending on their age, HLA-DQ genotype, islet autoantibody status and possible other factors.
– This was all new to me, and I enjoyed the whole working process. How the results from the statistical analyses opened up for new angels and questions and the continuous revision, that finally resulted in a finished product, Matilda Svensson concludes.
What is the Better Diabetes Diagnosis study group?
The Better Diabetes Diagnosis (BDD) study is a nationwide prospective cohort study that recruits new-onset diabetes patients who are less than 18 years old at time of diagnosis. Today are all paediatric medical centres in Sweden participating in the BDD study. The BDD study started recruiting new incident patients of type 1 diabetes from May 1st, 2005 and is still ongoing. Children who are less than 18 years old at the time of diagnosis were included, and the diagnosis is confirmed based on clinical criteria and follow up after six months of onset.
For each newly diagnosed child, blood samples are collected within 3 days of diagnosis for specific risk testing of HLA genotyping and islet autoantibody assays. At the same time blood samples are collected for biochemical and metabolic parameters, which are tested at the referring clinical centre and a specific nurse-based questionnaire is filled. The questionnaire records demographical variables as well as clinical and family medical histories, and also involves information about the patient’s origin for two generations.
New Insights into the Development and Maturation of Monocyte-derived dendritic cells.
F. Sprater, W. Azeem and S. Appel
In this paper, the group of Silke Appel for the first time describes that the expression of the transcription factor ESE-3 is regulated by the proliferator-activated receptor gamma (PPAR-γ) pathway. The group has previously shown that high ESE-3 expression correlates with the immunogenic functions of dendritic cells, and that tolerogenic dendritic cells have low ESE-3 expression. Dendritic cells have an outstanding ability to modulate immune responses and are therefore an interesting target for use in immunotherapy towards cancer and autoimmune diseases. Vaccines based on dendritic cells have shown promising results in mouse models. Monocyte-derived dendritic cells (moDCs) can be generated in vitro from peripheral blood mononuclear cells. A better understanding of the molecular mechanisms involved in moDC development and maturation is important to fully exploit the therapeutic potential of moDCs.
Florian Sprater was a PhD student in Silke Appel´s research group and this paper is included in his thesis. He was the one who performed all the pilot experiments and also carried out a lot of the final experiments.
– For me, the most fun with this project was the work in the lab, Florian says.
Florian Sprater and co-workers found two putative binding sites for PPAR-γ upstream of the ESE-3 gene. Therefore the effect of PPAR-gamma activation of ESE-3 expression in moDCs was investigated. They were able to show that ESE-3 expression is upregulated in moDCs, despite the tolerogenic phenotype of these cells. This could be due to the choice of method to generate the cells, indicating the importance of careful investigation of the phenotype and function of in vitro-derived cells before they are used clinically.
CD40L, an interesting target in autoimmune disease?
R. Belkhir, N. Gestermann, M. Koutero, R. Seror, J. Tost, X. Mariette and C. Miceli-Richard
This study shows that membrane-bound CD40L is overexpressed in ex vivo activated CD4+ T cells from females with the autoimmune disease primary Sjögren´s Syndrome (pSS). However, this overexpression did not involve epigenetic changes in key regulatory regions. This is in contrast to what has been reported previously for another autoimmune disease, systemic lupus erythematosus (SLE).
There is a female predominance of autoimmune diseases, which could be explained by epigenetic deregulation of genes encoded on the X chromosome. CD40L is one of the genes involved in adaptive and/or innate immunity that is located on the X chromosome.
X-inactivation and escape
X-inactivation is a process by which one of the two copies of the X chromosomes present in female mammals is inactivated. The inactive X chromosome is silenced by its being packaged in such a way that it has a transcriptionally inactive structure called heterochromatin. As female mammals have two X chromosomes, X-inactivation prevents them from having twice as many X chromosome gene products as males. However, some genes escape this inactivation, and many studies have shown that epigenetics plays a crucial role. It has also been shown that women with autoimmune diseases have more frequent X-inactivation escape than women without the diseases. CD40L is located on the long arm of the X chromosome and is a good candidate to asses this hypothesis.
No epigenetic changes found in pSS patients
However, despite the fact that epigenetic changes such as hypomethylation of regulatory regions of CD40L was found both in patients with SLE and systematic sclerosis, this was not found in this study. Still, the overexpression found here, and in other studies, suggests that CD40L could be an interesting target in autoimmune disease.