Humoral Rejection in Allografts And Gene Expression Anatasia Gangadin Dr. Mario C Deng Columbia University College of Physicians and Surgeons

Key Ideas Allograft Endomyocardial Biopsy Rejection CARGO Functional Genomics Patient Care

Endomyocardial biopsy Currently Only way to test for rejection Risk Factors associated

Rejection overall survival of heart transplant is 85% IncidenceMortality (Of those affected) Humoral17.6%17% Acute Cellular 1.4%23% Chronic13%18% Mild Severe

CARGO Study To Reduce the Number of Biopsies by taking samples of blood as opposed to heart Tissue Being able to predict rejection after a transplant Establishing a pattern of Genes which could ultimately predict Rejection

Basic Forms Of Rejection Chronic Rejection The movement of Smooth Muscle Tissue into the Coronary Arteries Causes Overtime narrowing of the Coronary Arteries Leads to lack of blood flow, Tissue Death, Heart Failure Cellular Rejection Inflammatory Response due to T cell infiltration in tissue Cardio Myocyte Damage Most Common Form Of rejection

Humoral Rejection Accounts for 20-30% of rejections in allografts. Common Risk Factors Causes hemodynamic dysfunction (shock, hypertension, decreased cardiac output, and rise in pulmonary artery pressure).

How Humoral Rejection Works

T Cell receptor CD 4 CD 8 HLA Antigen Displayed

Gene Expression When Lymphocytes release specific cytokines and plasma cells, They are doing so because of the patient’s gene Expression, which is why some patients experience rejection or no Rejection.

Experiment Aim: To compare genomic data in 121 patients and to discover a set of genes which can be used to predict Present/Future Humoral Rejection. Inclusion: Patients with Heart Transplants within past 6 months Patients >18 Exclusion: Patients with Oversensitive Immune Systems Patients > 1 transplant

Methods Gene Microarray Analysis CD4 Staining Gene Microarray and Statistical Analysis

Gene Microarray Analysis

C4D Staining

Genes GENENAMEFUNCTION CD28CD28 moleculeEssential for CD4 T cell proliferation CD40CD40 moleculeMediates in CD40 responses, inflammation, and other Immune responses C4BComplement component 4B Codes for Compliment Factor 4, provides interaction between antigen and compliment components CD8Compliment Component 8D Present on surface of T lymphocytes, Helps recognize Antigens displayed by Antigen Presenting Cells CCR6Chemokine receptor 6Helps B cell differentiation, and maturation. Expressed by CD4 and CD8 (natural killer cells) CCR4Chemokine (C-C motif) receptor 4 Codes for proteins which control development, homeostasis and function of Immune System CCr7Chemokine receptor 7Codes for proteins which activates B and T lymphocytes CCL2(C-C motif) ligandCodes for Cytokine Production which are responsible for inflammatory response IL2Interleukin 2Encodes for protein which releases a cytokine important for B and T cell proliferation IL10Interleukin 10Codes for cytokines which effects immunoregulation and inflammation, and B cell survival

Future Research To Perform Microarrays in order to pinpoint specific genes from patient blood To sample patient blood for CD4 which may detect rejection, instead of heart tissue

References Marboe C, Deng MC, Billingham M. Nodular Endocardial Infiltrates (Quilty Lesions) Cause A Significant Variability in Diagnosis of ISHLT Grade 2 and 3A Rejection in Cardiac Allograft Recipients. Journal of Heart and Lung Transplantation July 2005; 24:s219-s226. Evans R, Williams G, Deng MC. The Economic Implications of Noninvasive Molecular Testing for Cardiac Allograft Rejection. American Journal of Transplantation 2005; 5: Deng MC, Eisen HJ, Mehra MR. Noninvasive Discrimination of Rejection in Cardiac Allograft Recipients Using gene Expression Profiling. American Journal of Transplantation 2006; 6: Deng MC. Cardiac Transplantation. Heart 2002; 87: Michaels PJ, Fishbein MC, Colvin RB. Humoral rejection of Human organ Transplants. Springer Seminars in Immunopathology

Acknowledgements Dr. Mario Deng Martin Cadeiras Manuel Prinz von Bayern Sarfaraz Memon Dr. Sat Bhattacharya Columbia College of Physicians and Surgeons Memorial Sloane Kettering Cancer Center Harlem Children Society