Immunology of CELIAC DISEASE Ibrahim, Ibrahim Mentor: Dr. Sanil Manavalan Columbia University, College of Physicians and Surgeons

Celiac disease a digestive disease that damages the small intestine and interferes with absorption of nutrients from fooda digestive disease that damages the small intestine and interferes with absorption of nutrients from food it is a genetic disorderit is a genetic disorder some of the symptoms of celiac disease are diarrhea, weight loss, malnutrition, etc…some of the symptoms of celiac disease are diarrhea, weight loss, malnutrition, etc… when people with celiac disease eat foods containing gluten, their immune system responds by damaging the small intestinewhen people with celiac disease eat foods containing gluten, their immune system responds by damaging the small intestine Diagnosis is by intestinal biopsy histological grading and presence of serum auto- antibodies.Diagnosis is by intestinal biopsy histological grading and presence of serum auto- antibodies. because the body's own immune system causes the damage, celiac disease is considered an autoimmune disorderbecause the body's own immune system causes the damage, celiac disease is considered an autoimmune disorder

AUTOIMMUNE DISEASE: condition in which the immune system mistakenly attacks the body's own organs and tissues AUTOIMMUNE DISEASE: condition in which the immune system mistakenly attacks the body's own organs and tissues Type 1 Diabetes – affects the pancreasType 1 Diabetes – affects the pancreas Systemic Lupus erythmatotosus (Lupus)– affects the skinSystemic Lupus erythmatotosus (Lupus)– affects the skin Crohn’s Disease- affects the gutCrohn’s Disease- affects the gut Multiple Sclerosis- affects the brainMultiple Sclerosis- affects the brain

Key Terms Cytokines – proteins secreted by macrophages and leukocytes that stimulate or inhibit the proliferation or function of immune cells. –Interleukin – cytokines that only affect leukocytes. Lymphocyte – A type of leukocyte (white blood cell) of the immune system. –T-Regulatory Cell – slows down and stops immune response –T-Helper Cell – has antigen-receptors, releases cytokines –Natural Killer Cell – kills macrophage and bad stuff –Cytotoxic T-Cell – kills cells that produce foreign antigens such as cells infected by viruses –B-Cell – creates antibodies. Antigen – invading microbe Antibody – protein that fights bacteria and viruses. Lymphocytes

The Gut Lumen Villi- (singular: villus) are tiny, finger-like structures that protrude from the wall of the intestineVilli- (singular: villus) are tiny, finger-like structures that protrude from the wall of the intestine -Microvilli- hair-like structure on the surface of absorptive and secretoryepithelial cells -Microvilli- hair-like structure on the surface of absorptive and secretory epithelial cells Epithelial cells - layer of cells which line the GI tractEpithelial cells - layer of cells which line the GI tract Diagram

Glaidin Enterocytes GUT LUMEN Antibodies anti-glaidin anti-endomysial And tissue trasgliutaminase Cytokines (Interferon- , TNF-  IL-15 etc) Lymphocytes (T cells, Natural Killer cells and B cells) Plasma cells  T cell receptor Helper T cell HLA-DQ2/DQ8 molecule Antigen Presenting Cell GUT LUMEN LAMINA PROPRIA AND INTRA EPITHILIAL SPACE Inflammation & Damaged enterocytes

Objective To find the cytokine levels in the serum of patients in the following groupsTo find the cytokine levels in the serum of patients in the following groups –Healthy Patients (Control) –Active Celiac Disease Patients –Celiac Disease patients on Gluten-free diets –Refractory Disease Patients The following cytokines were checked in the serum of these patients;The following cytokines were checked in the serum of these patients; –IL-1 β –IL-6 –IL-8 –IL-12 p70 –IFN- γ –TNF- α

Methods Dilution of Standards Standards were used to calibrate the flow cytometer. The Standards were diluted by 1:1, 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, 1:125, 1:250, and 1:500. The standards were just FACS buffer solutions in water. Calibration of Cytometer To do this, we had solutions of the cytokines we were testing. After this we calibrated the equipment so it recognizes which wavelengths of light are representative of specific cytokines. Excel Spreadsheet To analyze the results, and present them to the scientific community, we used Excel spreadsheet to put the data into a comprehensible graph format.

Flow Cytometry DEFINITION: technique for counting, examining and sorting microscopic particles (such as cells) suspended in a stream of fluid ROLE IN OUR EXPERIMENT: To analyze concentrations of cytokines in the sera of patients using capture beads which detect the cytokines.

Hypothesis  Increased serum levels of inflammatory cytokines in patients with active and refractory celiac disease  Normal levels of cytokines in gluten –free diet

Results

Results Cont.

Conclusions Pro-inflammatory cytokines such as Interferon –gamma and IL-8 seemed to be increased in both patients with active disease and Celiac patients on a gluten-free diet. We are analyzing the data on an individual patient by patient basis to determine if there is a correlation with the biopsy grade for each patient

Acknowledgements Dr. Sanil Manavalan - Mentor Dr. Jamal Naiyer – Co-mentor Sindhu Kunduvaram – Undergraduate Student Dr. Sat Bhattacharya – Coordinator OF HCS Mrs. Susan Farenholtz – Coordinator of Project Seed Harlem Children Society Project Seed

Cytokines Cytokine IL-1 β IL-6 IL-8 IL-12 p70 IFN- γ TNF- α Principal cell Targets and biological effects Endothelial cells: activation B cells: proliferation of antibody producing cells Activator of neutrophils NK cells and T cells: IFN-gamma synthesis, increased cytolytic activity Macrophages: Activation Endothelial cells: Activation