Day 3 :
- Track 13: Neuro Immunology
Track 14: Reproductive Immunology
Track 15: Costimmulatory Pathways
Track 16: Diagnostic Immunology
Track 17: Technological Innovations in Immunology
Track 18: Immunoinformatics and Systems Immunology
Track 19: Rheumatology
Track 20 Nutritional Immunology
Track 17: Technological Innovations in Immunology
Track 18: Immunoinformatics and Systems Immunology
Track 19: Rheumatology
Track 20 Nutritional Immunology
Location: Harbor Room
University of Nebraska Medical Center, USA
Ligia A Pinto
Frederick National Laboratory for Cancer Research, USA
Title: In Pursuit of the Unknown: Standardizing Endpoint Assays for Evaluating Complex Immunological Signatures
Time : 09:00-09:20
David Hokey is the Sr. Director of Immunology & Animal Studies at Aeras, a non profit biotech advancing the development of new tuberculosis vaccines. He received his Ph.D.in Immunology from the University of Pittsburgh in 2005 where he worked on dendritic cell-based tumor vaccines. He then performed his postdoctoral research with Dr. David Weiner at the University of Pennsylvania,focusing on DNA vaccines for HIV, cancer, and influenza with an emphasis on primate immunology and polychromatic flow cytometry. Dr. Hokey’s current work at Aeras involves evaluation of immune responses to TB vaccine candidates in pre-clinical and clinical settings. His research interests include evaluation of novel vaccine platforms and adjuvants for manipulation of immune responses.
BacilleCalmette-Guérin (BCG) remains the only licensed vaccine for tuberculosis (TB). While BCG has been shown to be effective in preventing certain forms of childhood TB, adult pulmonary TB continues to be a serious burden, with 8.6 million new cases and 1.3 million deaths reported in 2012. Despite the availability of drug therapies, TB remains one of the leading causes of death by an infectious disease worldwide. Furthermore, the emergence of drug-resistant strains of Mycobacterium tuberculosis (Mtb) emphasizes the need for a new and effective vaccine. However, TB vaccine development has been hindered by suboptimal and expensive animal challenge models in the face of limited funding, the lack of a human challenge model, and a lack of a correlate of immunity or protection. In addition, immune assays used in evaluating vaccine candidates are variable, particularly assays measuring cellular responses. Here we discuss some of the issues with cellular assays and show the results of the qualification of a 13-color intracellular cytokine staining (ICS) assay that includes the detection of phenotypic markers, memory markers, and 7 T-cell functions (IFN-γ, IL-2, TNF, IL-17, IL-22, CD107a, and CD154). We show that at least partial qualification of this complex assay is feasible and propose a useful method for assessing assay variability and performance over time for cellular assays used in clinical trials.
University of Nebraska Medical Center, USA
Title: Circulating plasmablastsfrom patients with rheumatoid arthritis preferentially produceanti-citrullinated proteinantibodies
Time : 09:20-09:40
Kaihong Su obtained her doctorate degree and post-doctoral training from the University of Alabama at Birmingham (UAB). After being a faculty member at UAB for 5 years, she joined the University of Nebraska Medical Center in 2008 to continue her research. The long-term goal of her lab is to understand the molecular mechanisms for autoimmune diseases, including SLE and RA. She has published 26 papers in reputed journals and served as ad-hoc reviewers for a number of scientific journals.
Anti-citrullinated protein antibodies (ACPAs) are highly specific serological markers for rheumatoid arthritis (RA) and are also believed to contribute to RA pathogenesis. However, the cellular and molecular basis of ACPA production is not fully understood. Using a single cell PCR-based antibody cloning technology, we analyzed ifcirculating plasmablasts(CD19dimCD27highCD20-) inRA patientsproduce ACPAs.Among the 195 antibodies generated from6ACPA+RA patients, 18.5% (ranging 6.5-27.6%) specifically recognize citrullinated antigens, but not their native forms. However, none of the antibodies generated from ACPA-RA patientor healthy donors reacted with citrullinated antigens. Detailed sequence analyses showed that the IgH and IgL genes encoding these ACPAs are highly mutated. Reversion of the mutated IgH and IgL genes to theircorresponding germlinegenes completely eliminated their ACPA reactivity, suggesting that the generation of circulating ACPAs is an antigen driven process. To identify the antigen sourcesfor ACPAs, we found that about half of the ACPAsreact with outer membrane proteins from Porphyromonagingivalis(P. Ging) and/or citrullinated P. Gingenolase. Some ACPAs also react with auto-antigens released in the human neutrophil extracellular traps. These results suggest that circulating plasmablasts in RA patients are a source of ACPA production and the generation of ACPAsis driven by both exogenous bacterial antigens and citrullinated self-antigens. Our data thus identifycirculating plasmablastsas potential therapeutic targetsand provide a mechanism for previous findings that plasmablasts in the bloodpredictnonresponse to anti-CD20therapy in RA patients.
Halozyme Therapeutics Inc., USA
Time : 09:40-10:00
John Huang has his career as a drug development biologist in pharmaceutical companies, focusing on oncology, metabolic disease and immunology. Currently, he is working at Halozyme Theraprutics Inc., leading the cell biology group as Associate Director. His research and preclinical work focus on understanding the pharmacological mechanism of action of PH20, as well as exploring hyaluronan biology in all applied disease areas. He was a senior principal scientist at Pfizer Inc., a senior Scientist at Receptor Biologix Inc., following being a Scientist at Amgen Inc. He graduated with honors from the University of California Riverside, and had his Postdoc training at UCSF with Dr. J Michael Bishop.
Hyaluronan (HA) is involved in many biological activities, such as inflammation, angiogenesis, and tumorigenesis. Some activities are associated with specific sizes of HA molecules, the degradation intermediates of hyaluronidases, such as PH20. This enzyme cleaves HA and generates lower molecular weight (LMW) HA, which was reported to stimulate the innate immune response. However, most PH20 enzymes used were protein extracts from animal tissues, e.g., bovine testis-derived hyaluronidase (BTH), and prone to artifacts due to impurity (including endotoxin). We tested a highly purified recombinant human hyaluronidase, rHuPH20, in the air pouch model of inflammation. In our study, rHuPH20 degraded HA to LMW fragments ranging from 50-500 kDa. While lipopolysaccharide (LPS) stimulated many cytokines and chemokines, rHuPH20 did not stimulate any cytokines and chemokines, suggesting that generation of LMW HA in situ does not stimulate cytokine and chemokine production. LPS also induced neutrophil infiltration into the air pouch, which was not observed with rHuPH20 treatment. Exogenous LMW HA failed to stimulate either cytokines and chemokines or neutrophil infiltration. BTH contain endotoxin and stimulated a strong inflammatory response, which was largely reduced by endotoxin removal. When dosing rHuPH20 together with LPS, the profiling of cytokine and chemokine stimulation was the same as LPS treatment, but neutrophil infiltration was inhibited. The rHuPH20-mediated anti-neutrophil infiltrating effect was also observed in LTA-induced neutrophil infiltration. In conclusion, our results indicate that neither rHuPH20 nor LMW HA has inflammatory properties, and instead, rHuPH20 can inhibit some aspects of inflammation, such as neutrophil infiltration into the air pouch.
Leidos Biomedical Research, Inc., USA
Time : 10:00-10:20
Martha Sklavos earned her undergraduate degree from Johns Hopkins University andher PhD in Immunology from the University Of Pittsburgh School Of Medicine. Currently, Martha is a post-doctoral fellow at the HPV Immunology Laboratory at the Frederick National Laboratory for Cancer Research where she was awarded a grant from the National Cancer Institute to investigate a novel biomarker for cervical cancer risk. She is now leading several studies to define the role of this novel biomarker in additional cancer types. Martha enjoys conceptualizing ideas and driving projects forward and hopes to fill a similar role in the pharmaceutical industry upon completion of her post-doctoral training.
Fanconi anemia (FA), dyskeratosiscongenita (DC), and Diamond-Blackfan anemia (DBA) are rare inherited bone marrow failure syndromes (IBMFS) characterized by birth defects, bone marrow failure, and increased frequency of malignancy.Females with FA are known to have a high incidence of infertility which may be associated with primary ovarian insufficiency (POI), while infertility in DC and DBAhas not been reported.Anti-Müllerian hormone (AMH) is a marker of ovarian reserve and the decline of AMH is correlated with the loss of fertility as AMH decreases to undetectable levels five to ten years prior to menopause.In this study, we examined whether serum levels of AMH could serve as a menstrual cycle-independent marker for the diagnosis of POI in patients with IBMFS. Serum AMH levels were measured in female participants in the National Cancer Institute’s IBMFS cohort in 22 FA patients (median age 15 years, range 7-37), 20 unaffected FA relatives (age 33.5, range 3-40), 15 patients with DC (age17, range 4-32), 18 unaffected DC relatives (age22.5, range 2-40), 12 patients with DBA (age15.5, range 1-30), 13 unaffected DBA relatives (age11, range 1-34), and 21 unrelated healthy females (age27, range 12-40). FA females had very low or undetectable AMH levels (median 0.05 ng/ml, range 0-2.32ng/ml) compared with bothunaffected FA relatives (2.10 ng/ml, range 0.04-4.73ng/ml, p<0.0001) and unrelated healthy females (1.92ng/ml, range 0.31-6.64 ng/ml, p<0.0001).All five females with FA over the age of 25 had undetectable AMH levels and were diagnosed with POI.Six of seven FA patients over the age of 20 developed cancer (median age 27; three prevalent, three incident) while all unaffected female relatives and unrelated controlswere cancer-free.Femaleswith DC had AMH levels that were significantly lower(0.55 ng/ml, range 0.01-4.82 ng/ml) than their unaffected relatives (2.33ng/ml,range0.44-10.03 ng/ml, p=0.001) and unrelated healthy controls (p=0.001) while AMH levelsof females with DBA (0.90 ng/ml, range 0-10.20) did not significantly differ from their unaffected relatives (1.71 ng/ml,range: 0.56-5.10 ng/ml, p=0.21) or unrelated healthy controls (p=0.14).One patient with DC reported fertility issues, two patients with DBAwere diagnosed with POI,and all three had AMH levels under 1ng/ml.Only one patient with DC hadprevalent cancer while DBA patients and all relatives ofpatients with IBMFS were cancer-free. This study is the first to demonstrate AMH deficiency in females with IBMFS and to suggest that measurement ofAMH is useful for the early diagnosis of POI in IBMFS.Our data suggest that AMH may be lower in FA than in DC and DBA.Prior studies indicate that the rates of cancer are highest in FA, followed by DC, and lowest in DBA.Further researchis warranted to determine whether there is an association between AMH deficiency and increased cancer risk in IBMFS or within the general (unaffected) population.
University of Newcastle, Australia
Time : 10:20-10:40
Andrea’s research interests comprise of: paediatric rheumatology, podopaediatric, biomechanics, orthotic management, MSK and health in Aboriginal and Torres Strait Islanders. As part of his PhD study, Andrea has carried out a multicentre randomised controlled trial (RCT) to investigate the clinical effectiveness of foot orthoses on pain, quality of life and gait dynamics of patients diagnosed with Juvenile Idiopathic Arthritis (JIA). This published work is providing new research evidence for podiatrists working within the paediatric rheumatology multidisciplinary team. Andrea has presented his work at several national and international conferences and has been actively involved in providing podiatric care in different hospitals and private clinics for professional athletes and children. Andrea is originally from Italy, he worked as a lecturer and researcher at Queen Margaret University in Edinburgh (UK). After nearly 10 years living in Scotland, in July 2013 he moved to Australia to commence a new exciting lecturing post for the podiatry department at the University of Newcastle (NSW). Ciao
Introduction: There is limited evidence supporting the podiatric treatment of children with juvenile idiopathic arthritis (JIA). This multicentre randomised controlled trial aimed to determine whether pre-formed foot orthoses (FOs) impacted on pain and quality of life (QoL) in children with JIA. Methods: Eligible children were randomised to receive either “fitted” FOs with customised chair-side corrections or “control” FOs made without corrections. Changes in pain and QoL were measured using a visual analogue scale (VAS) and PedsQL questionnaire respectively. JIA children were assessed at baseline, 3 months and 6 months. Results: Sixty children were recruited. One-hundred and seventy-nine out of a possible one-hundred and eighty assessments [99.4%] were completed. A statistically significant greater difference in pain reduction (baseline – 6 months) was seen between the two groups favouring fitted FOs (p = 0.029). The reduction in pain in the fitted FOs group was clinically important (8mm). Significant differences in QoL favouring fitted FOs were also identified as measured by the children and independently by their parents/carers. Conclusion: Fitted FOs may reduce pain and improve quality of life in selected children with JIA.
Columbia University School of Nursing, USA
Time : 10:55-11:15
Margaret A. Romero NP-C is a board certified nurse practitioner and graduate of Columbia University School of Nursing. She received her undergraduate Biology degree from SUNY Stony Brook. Ms. Romero has served as a preceptor and didactic instructor to physician assistants and nurse practitioner graduate students from Columbia University. As the founder of From Lupus to Living, her passion for working with individuals afflicted with lupus stems from her own personal journey with lupus nephritis. Her unconventional protocol has kept her out of the hospital and off the kidney transplant list.
People die everyday from the complications of lupus and more than 5 million worldwide are afflicted. The pain and fatigue have forced countless individuals into disability. Simple dietary modifications can create significant changes. Discussed are vital aspects of nutrition with the use of functional medicine to lessen and alleviate the symptoms of lupus. Topics includetesting for vitamin deficiencies, hormone levels, and gut function. Lab values with how to treat these deficiencies will be demonstratedalong with the proper use of vitamin supplementationas well as treatment of chronic underlying infections.Food allergies and anti-inflammatory foods are also reviewed.Until now, medications have been the mainstream treatments for lupus. Nutrition,functional testing, and nutraceuticals can be additional assets to modern medicine.
Instituto Nacional de Perinatología, Mexico
Title: The role of microRNAs and CD4+ T lymphocyte subsets in the regulation of the immune system during normal pregnancy
Time : 11:15-11:35
Estibalitz Laresgoiti-Servitje is a researcher at the National Institute of Perinatology (INPer) and an associate professor at the Monterrey Institute of Technology and Higher Education (ITESM), in Mexico City. She completed a Medical Doctorate, a Masters in Science in Immunology, a Masters in Neurosciences, and a PhD in Health Psychology. Her research interests include the neuroendocrine regulation of the immune system and the modulation of the immune system during normal pregnancy and preeclampsia.
miRNAs are non-protein-coding RNAs that regulate gene expression and may play a role in changes that occur during the gestational period. Some miRNAs can be involved in implantation, some may be expressed in the placenta under hypoxic conditions, and others may participate in the induction of immune tolerance during the gestational period. Several miRNAs have been identified as pregnancy-associated. These miRNAs could behave as diagnostic biomarkers as some may participate in pregnancy disorders like preeclampsia and fetal growth restriction. The role of miRNAs in pregnancy is not fully understood yet. CD4+ T lymphocyte subsets are key players in the regulation of the immune system during normal pregnancy. The Th1/Th2 paradigm has been used to explain T cell behavior during pregnancy for many years. However, the sole use of this model to elucidate the role of T lymphocytes in the regulation of the immune system during the gestational period could be over-simplifying the mechanisms involved. The participation of regulatory T lymphocytes (CD4+CD25+FoxP3+ Tregs) is essential for pregnancy continuation. Furthermore, four different Treg subsets have been identified throughout pregnancy, which differ on their expression of CD45RA and HLA-DR. The proportion of these subsets may participate in the development of certain pregnancy complications. Lastly, Th9 and Th2 cells may influence these Treg subsets. Research on the interactions between CD4+ lymphocyte subsets and Treg subsets during normal pregnancy may broaden our perspective regarding the regulation of the immune system during the gestational period, and may help us understand the pathophysiology of some pregnancy-related diseases.
Frederick National Laboratory for Cancer Research, USA
Time : 11:35-11:55
Ligia Pinto completed her PhD in 1995 at the Experimental Immunology Branch, National Institutes of Health, after several years of research on cellular immunology of HIV infection at the University of Lisbon, School of Medicine in Portugal. She continued her postdoctoral studies at NIH where she focused on investigating immunological alterations induced by HIV as well as host protective immune responses that may control HIV replication and associated immunopathogenesis. In 2001, she became the head of the HPV Immunology Laboratory at the Frederick National Laboratory, where she is a Principal Investigator and leads a team of scientists, postdoctoral fellows, research associates and students. She has worked in the area of immunology of infectious diseases and vaccines for the last 27years, with over 80 peer-reviewed publications, has many invited international presentations and received a number of distinguished scientific awards.
There are two licensed prophylactic human papillomavirus (HPV) vaccines, aquadrivalent that protects primarily against HPV-16, HPV -18, HPV-6 and HPV-11and a bivalent vaccine that protects primarily against HPV-16 and HPV-18. These L1 VLP vaccines have proven to be very efficacious in preventing infections and pre-cancerous lesions at the cervix caused by HPV-16 and HPV-18. Furthermore, partial protection against phylogenetically-related oncogenic types not included in the vaccine has also been reported. Although correlates of vaccine protection have not been identified, neutralizing antibodies are thought to be the primary mechanism of vaccine induced protection against infection. HPV vaccines induce high titers of anti-L1 IgG antibodies, to the HPV types included in the vaccine, that persist for several years after vaccination at levels considerably higher than those observed in natural infection. These antibodies are detected not only inserum but also at the cervix, where local antibody levels correlate well with serum levels. L1 VLP vaccines also induce neutralizing antibodies tophylogenetically-related types not included in the vaccine. These can also persist for years after vaccination but withtiters much lower than those observed for the homologous vaccine types. Antibody avidity increases with each dose of vaccine, with affinity maturation occurring after the 3 doses of vaccine. Interestingly, levels of avidity did not appear to correlate well with antibody titers. Despite all the information on antibody responses to HPV vaccines, future efforts are needed to standardizemethods and tools for evaluation of immunogenicity of HPV vaccines and to better identify mechanisms of immunogenicity and correlates of protection against infection.
Institute of Health Research Gregorio Marañón, Spain
Title: HIV infection of Human regulatory T cells (Treg) downregulates Foxp3 expression and produces a loss of the suppressive capacity of these cells
Time : 11:55-12:15
Rafael Correa Rocha has completed his first PhD in Biology at the Universidad Complutense of Madrid (Spain) in 2004. His thesis was awarded with the National Prize of Doctorate 2005. He joined the ISREC (Epalinges, Switzerand) as a post-doctoral researcher and afterwards, he obtained a position as Assistant Professor at the HopitauxUniversitaires de Genève (Switzerland). He joined the IISGM of Madridas a Senior Scientist in February 2008. He completed a second PhD in Medicine at the Universidad Autónoma of Madrid in 2014. At present, he is the Head of the Laboratory of Immune-regulation at IISGM. He has published more than 40 papers in reputed journals.
Regulatory T cells (Treg) play an important role in infections modulating host immune responses and avoiding over-reactive immunity. Immune hyperactivation associated with HIV infection lead to a marked erosion and deregulation of immune system, and by that, the role of Treg in HIV-infected patients is critical because their implication preventing this hyperactivation. The findings about the role of Treg in HIV infection are controversial, and considering that Treg are susceptible of being infected by HIV, there are not data about the effect of HIV infection on Treg phenotype and function.We demonstrated by first time that HIV infection of Treg markedly disturb the phenotype of these cellsdownregulating the expression of Foxp3 and CD25, which is followed by a loss of their suppressive capacity . We also demonstrated that the balance between Treg and effector cells is broken in HIV patients by a direct effect of the virus on Treg, and finally we have also described that HIV-infected patients have a marked deficit and impaired function of Treg that would be related with the incidence of Immune hyperactivation in these patients.
Universidad Austral de Chile, Chile
Time : 12:15-12:35
Alejandro J Yanez Carcamo has completed his PhD at the age of 32 years from Universidad Austral de Chile University and doctoral studies from UMKC, U.S.A. He is the director of Austral Omics, a service organization that provide a transversal solution to the researchers. He has published more than 60 papers in reputed journals contributing to the knowledge of metabolism and applied microbiology.
Diabetic nephropathy (DN) is one of the major complications of diabetic patients and is the leading cause of end-stage renal disease. Kidneys are major sites of deregulated glucose production in diabetic patients. It is therefore crucial to investigate the mechanisms involved in their pathogenesis. We analyzed theexpression of the insulin receptor (IR) in renal cortex and proximal tubules from diabetic rats and humans. Surprisingly, in the kidney of both human type 2 diabetic patients and in a type 1 diabetic rat model, a significant reduction in the protein levels of IR and a consequent increase of PEPCK is produced.Thus, expression of IR protein in proximal tubules from type 1 and type 2 diabetic kidney indicates that this is a commonregulatory mechanism which is altered in DN, triggering enhanced gluconeogenesis. Moreover, we detected activation of the GSK3βkinase andoverexpression of muscle glycogen synthase (MGS)and glycogen deposition in the diabetic kidney from rat and human, that correlate with induced-cell death in a model of human renal tubule cells. This differential expression suggests the participation of MGS in the renal metabolic changes associated that also are induce the increased levels of inflammation markers ( MCP-1, ICAM-1) and activation of NFk-B.Hence, alteredexpression of IR in human diabetic kidney could be one of the maintriggers for enhanced inflammation and gluconeogenesis and the consequent kidneydysfunction in DN. (FONDECYT 1131033.)
National Institute on Aging, NIH, USA
Time : 12:35-12:55
Catalina Lee-Chang has completed her PhD in Immunology from University of Lille Nord de France in 2010. Her work focused on the role of B cells in multiple sclerosis pathogenesis and neuroinflammatory mechanism. In 2011 she joined Arya Biragyn’s lab (NIH) as postdoc to continue her training in B-cell biology, as well as cancer and aging immunology.
CD8+CD28- T cells are considered an immune risk of mortality and morbidity in elderly. The mechanism of their enrichment and their function in elderly is poorly understood. In the present study we report a novel and evolutionary-conserved B-cell subset expressing 4-1BBL, which accumulates with aging of mammals, such as humans, rhesus macaques and mice. These cells (termed 4BL cells) induce antitumor effector CD8+ T cells by presenting endogenous antigens and utilizing 4-1BBL/4-1BB axis. 4BL cells are also responsible for the expansion of auto-reactive CD8+ T cells in patients treated with high dose chemotherapy and autologous progenitor cell transplantation. Thus, accumulation of antitumor 4BL cells explains the paradox of retarded tumor growth in the elderly, and most likely autoimmune disease outcome in the aged population. However, 4BL cell accumulation and their effects on CD8+ T-cell compartment can be eliminated by inducing B-cell lymphopoiesis in old mice (after B-cell depletion), suggesting that the aging-associated skewed cellular immune responses are reversible. We propose that 4BL cells and their 4-1BBL signaling pathway are useful targets for improved effectiveness of natural antitumor defenses and therapeutic immune manipulations in the elderly. How 4BL are generated, and their impact in the B-cell effector functions homeostasis in elderly will be further discussed. This research was supported entirely by the Intramural Research Program of the NIH, National Institute on Aging
University of Oslo, Norway
Time : 12:55-13:15
Geir Hetland is MD, PhD from University of Tromsø, Norway, and also American ECFMG graduate. He did his postdoctoral studies at The Scripps Research Institute, (1991-1993) when he was awarded the Fogarty International Award. He became Specialist in Immunology and Transfusion Medicine in 1995. He has studied and published on immunomodulatory and medicinal properties of mushrooms for over 15 years at The Norwegian Institute of Public Health and at Oslo University Hospital, where he is Senior Consultant and Professor of Immunology. He is also co-founder of Immunopharma, which aims at developing the Agaricus blazei-based extract, AndosanTM, into add-on hospital medicine.
Agaricus blazei Murill (AbM) is an edible Brazilian mushroom that has been used in traditional medicine for a range of diseases. It has been shown to have anti-infection and anti-tumor properties in the mouse, which are due to induction of Th1 responses. On the other hand, IgE-mediated allergy is induced by a Th2 response. Since according to the Th1/Th2 paradigm an increased Th1 response may promote a reduced Th2 response, the aim was to examine whether AbM had anti-allergy effects. A mouse model for allergy was employed, in which the mice were immunized s.c. with the model allergen ovalbumin (OVA). Additionally, the animals were given a mushroom extract, AndoSan, mainly (82%) containing AbM, but also Hericium erinaceum (15%) and Grifola frondosa (3%), or PBS p.o. either a day before or 19 days after the immunization. The mice were sacrificed on day 26, and anti-OVA IgE (Th2 response) and IgG2a (Th1 response) antibodies were examined in serum and Th1, Th2 and Treg cytokines in spleen cells cultures. It was found that the AndoSan extract both when given either before or after OVA immunization reduced the levels of anti-OVA IgE, but not IgG2a, in the mice. There was a tendency to reduced Th2 relative to Th1 cytokine levels in the AndoSan groups.
Afe Babalola University, Nigeria
Title: Haemogram and serum enzymes activities of Newcastle disease virus challenged broiler chickens following supplemental treatment with Aloe vera extract
Time : 14:00-14:20
Tony Ifeanyi Ojiezeh obtained his Ph.D from University of Benin, Benin City. He is a Biomedical Scientist of high repute and Laboratory Manager before join academics in 2008. He is the immediate past Head of Department and has published more than 20 papers in learned scientific journals. He is a Senior Lecturer in Afe Babalola University, Ado- Ekiti, Nigeria.
Aloe products have been promoted for use in constipation, coughs, diabetes, arthritis, immune-system deficiencies and many other conditions, but the clinical efficiency in chickens is unknown. Therefore, the study was designed to determine the haemogram and serum enzymes activities of Newcastle disease virus (NDV) challenged broilers following supplementation of Aloe vera. One hundred and forty day-old broilers were grouped into seven, 20 broilers/group. They were treated with different concentrations of the extract for 30 days. Both vaccinated and unvaccinated groups were challenged with 0.2 saline suspension of 106 ELD50 intradermal inoculation of NDV challenged strain on the 30th day. Increase in serum protein, aspartate transaminase, creatinine, urea and gamma globulin levels among challenged group was concentration dependent (50 mg > 100 mg > 150 mg) but not statistically significant (P > 0.05). Lymphocytosis among supplemented and challenged group was concentration dependent (50 mg > 100 mg > 150 mg) and statistically significant (P < 0.05). Control group lymphocyte count was within normal range (55 – 70 %). Heterophil and lymphocyte (H / L) ratio was lower in supplemented and challenged group. Oral intake of Aloe vera modulated leucocytes proliferation of the broilers and enhanced cell differentiation in favour of lymphocyte. Serum enzymes activities in broilers challenged with NDV following supplementation with A. vera juice were positively influenced, and modulated the excessive leakage of protein, globulin, creatinine and alkaline phosphatase in infected birds. Aloe extract may therefore be seen as a good immunomodulator.
Aligarh Muslim University, India
Title: Autoantibodies in rheumatoid arthritis may arise from immunogenicity of peroxynitrite-modified IgG
Time : 14:20-14:40
Yasir Arfat is pursuing his Ph.D. in Biochemistry at Aligarh Muslim University, India. He has been working to understand the role of AGE-IgG and its peroxynitrite-modified counterpart in the induction and/or progression of rheumatoid arthritis. He is a recipient of doctoral fellowship award on the basis of qualifying national level competitions. He has participated in several national and international conferences including one held at Budapest, Hungary. He is a member of the “World Society Interdisciplinary Anti-Aging Medicine (WOSIAM)” France.
Peroxynitrite (PON) is a potent oxidizing and nitrating agent with a biological half life of approximately 10ms. It is produced in vivo by diffusion-controlled reaction of nitric oxide and superoxide anion. It can oxidize and/or nitrate many amino acids causing changes in protein structure and function. The changes may lead to the pathogenesis of several inflammatory diseases including Rheumatoid Arthritis (RA). In this study, IgG was isolated from healthy subjects’ sera on protein A-agarose affinity column and PON was synthesized by rapid quenched flow method. PON-modified IgG (PON-IgG) was prepared by incubating IgG with PON at 370C for 30 min and maintaining pH at 10-11. Physicochemical alterations in PON-modified IgG were monitored by UV, fluorescence, CD and FT-IR spectroscopy, and SDS-PAGE. Oxidation and aggregation were assessed as free thiols, protein carbonyls, and thioflavin T and congo red binding. Nitrotyrosine, dityrosine and nitrotryptophan were also quantified. Formation of 3-nitrotyrosine was verified by LC-MS and HPLC, and attachment of PON to IgG was elucidated by MALDI-TOF mass spectrometry. PON-modified IgG exhibited hyperchromicity at 278 nm along with bathochromic shift and appearance of a new peak at 420 nm, decrease of tryptophan and tyrosine fluorescence, loss in β-sheet and appearance of new peak in FT-IR, compared to native IgG. SDS-PAGE results revealed concentration dependent decrease in the band intensity of PON-IgG compared to native IgG. Experimentally induced antibodies against PON-IgG showed high titre antibodies and specific binding may be due to generation of highly immunogenic neo-epitopes on PON-IgG. PON-IgG binding with circulating autoantinbodies of RA patients were compared with the experimentally induced antibodies against PON-IgG and its significance was analysed using biostatistical tools. The results so far suggest likely involvement of PON-IgG as an autoantigen in the induction and/or progression of RA.
University of Calcutta, India
Title: Leishmania donovani negatively regulates TLR4 mediated host response by preventing ubiquitination dependent degradation of TRAF3
Time : 14:40-15:00
Purnima Gupta is pursuing her PhD in Biochemistry from Calcutta University. She has received the prestigious CSIR-fellowship for doctoral studies. Her research interest includes understanding how intramacrophage pathogens evade host cell defences utilizing host signaling molecules to aid their survival within macrophage hostile environment. To elucidate this they chose visceral leishmaniasis as the disease model and L. donovani as a model organism. Till date they have unveiled how the parasite modulates TLR signaling pathway by means of upregulating the negative regulatory molecule, TRAF3. Moreover, they observed how the parasite regulates host molecule SOCS to inhibit host cell apoptosis and thereby save its niche for replication and propagation.
Leishmania donovani, causative agent of fatal visceral leishmaniasis, is an intramacrophage pathogen. It escapes host immune response by subverting Toll like receptor (TLR) signaling, which is critically regulated by protein ubiquitination. Our work identified tumour necrosis associated factor (TRAF) 3 as a target used by Leishmania to deactivate LPS-mediated TLR4 signaling. TRAF3 is an E3 ubiquitin ligase of the membrane associated signaling complex that regulates TLR pathway through distinct protein ubiquitination at specific residues. We observed that TRAF3 which is ubiquitinated at lys 48 position and subsequently degraded following LPS treatment, persisted in L. donovani as well as L. donovani + LPS co-stimulated cells due to defective lys 48 ubiquitination. Unlike lys 48, lys 63-linked ubiquitinated proteins has been implicated in the signaling mediated activation of the molecules. Our results revealed lys 63-linked ubiquitination of upstream proteins in the cascade (cIAP1/2 and TRAF6), mandatory for lys 48 linked TRAF3 ubiquitination and subsequent degradation was significantly reduced during infection. The reason may be reduced association between ubiquitin conjugating enzyme Ubc13 and TRAF6 during infection. Persistence of TRAF3 resulted in stabilization of signalosome complex at the membrane resulting in inhibition of TAK-1 mediated pro-inflammatory responses. Inhibition of TRAF3 prior to infection by small hairpin RNA in Balb/c mice also showed enhanced production of IL-12 and TNF-α and significantly decreased spleen and liver parasite burden. Our findings identified TRAF3 as a novel molecular regulator exploited by Leishmania for successful infection.
University of Mysore, India
Title: Cross talk between apoptosis, autophagy and necrosis – pathways involved in treatment of neuroblastoma, an in - vitro study
Time : 15:00:15:20
Shailasree Sekhar received her PhD from the CSIR Institute, Central Food Technological Research Institute at Mysore, Karnataka, India, in 2000. The thrust area under the Institution of Excellence was identified the biodiversity of Western Ghats medicinal plants (MP), with immunological affections and cancer prevention properties, due to location advantage of this hot spot to the University. She has been actively involved in compilation of their scientific data as reviews. Recently, she has brought out a database on medicinal plants of Western Ghats in an efficient way. Screening of MP with immunological affections used by tribes has resulted in identification of several of them with inflammation/cancer inhibiting property. Fingerprinting their metabolites has been her priority. She has published more than 30 papers in peer-reviewed journals, has 2 patents and is an adhoc reviewer of various journals. She has to her credit grants from National scientific agencies under Government of India.
Immunology ranks as one of the most rapidly expanding areas of biomedical science, covering topics from cancer and vaccine research to immune deficiencies and autoimmunity. Neuroblastoma and solid extracranial cancer of childhood with an annual incidence of ~ 9.1 cases per million children is reported. Standard treatments include radiation along with chemotherapy, however results in side effects to children due its acute toxicity. Advances in technology has allowed for search of new nontoxic drugs to be used as a single or multidrug therapy in natural products with almost 60% of drugs approved for cancer treatment being from natural origin. Apoptosis, defined as a controlled suicide program to remove defective cells, without damage to neighboring cells, acts via death receptor (extrinsic) and the mitochondrial (intrinsic) pathway. The mitochondrial pathway, involving loss of mitochondrial membrane potential (Δψm), is regulated through a highly intricate cross-talk, includes caspases 8, 9 and 3 situated at pivotal points of the signaling process. Caspase-8 studies represent a yet unknown pro-apoptotic pathway activated by cytotoxic drugs, and further, it was identified as an important p53 target gene in drug-induced death of cancerous cells. Autophagy, type-II cell death, a catabolic process, is phenotypically characterized by formation of large intracellular vacuoles, termed autophagosomes. It includes sequestration of cellular proteins and organelles in a specific sequence prior to the destruction of the nucleus and has been recognized in various cancer cell types as a response to anticancer therapies. Induction of autophagy, results in post-translational processing of cytosolic-associated microtubule-associated protein-1A/1B light chain 3 (LC3-I; ~17kDa), to autophagosome membrane-bound LC3-phosphatidylethanolamine conjugate (LC3-II; ~19 kDa), leading to formation of autophagosomes. p-coumaric acid (p-CA), a ubiquitous plant phenolic acid, has been proven to render protection against pathological conditions. Studies on p-CA as evaluated for its capacity to induce cytotoxic effect to neuroblastoma, N2a, cells and possible mechanism of its action will be discussed.
EFS Rhône-Alpes, France
I am an M.D/Ph.D in Biology. I work currently in a laboratory of a Health Clinical Center, CHU of Lyon. I specialize in immuno-haematology as well as in NIPT: fetal RHD genotyping on maternal plasma. I was invited to present this at several international conferences (ESHG 2012) and I am the author of a recently published article (Non-invasive fetal RHD genotyping: Validation of the method with 200 patients, TCB, 2014 ).
Background: Non invasive fetal RHD genotyping is an important tool to assess the risk of fetuse’s hemolytic disease of anti-D allo-immunized pregnant women by non invasive method Method: A method of genotyping has been developed in the laboratory of Lyon-GHE according to Minon’s team: Exon 4, 5, and 10 are amplified by real time PCR. At first, genotyping results of 200 pregnant women have been compared with RH1 phenotype at birth. Results: The most important parameters of validation have been tested: The sensibility and the specificity; the negative predictive value; the correlation study permitted to define criteria of biological interpretation. The validation of this method permitted to determine critical points and the limits of the method due to the minor amount of fetal DNA in the maternal plasma and existence of many variant forms of the RHD gene. Conclusion: Fetal RHD genotyping can optimize the obstetrical management of women RH -1 allo-immunized by a simple, reliable and minimally invasive method