Humoral Rejection in Allografts And Gene

ExpressionAnatasia GangadinDr. Mario C Deng

Columbia University College of Physicians and Surgeons

Key Ideas• Allograft

• Endomyocardial Biopsy

• Rejection• CARGO

• Functional Genomics

• Patient Care

Patient Care

Endomyocardial biopsy

• Currently Only way to test for rejection

• Risk Factors associated

Rejection

overall survival of heart transplant is 85%

Incidence

Mortality(Of those affected)

Humoral 17.6% 17%

Acute Cellular

1.4% 23%

Chronic 13% 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 RejectionChronic 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

Antibody Mediated 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 CD4

CD8

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 >18Exclusion: • Patients with Oversensitive Immune Systems• Patients > 1 transplant

Methods

• Gene Microarray Analysis• CD4 Staining

• Gene Microarray and Statistical Analysis

Gene Microarray Analysis

C4D Staining

GenesGENE NAME FUNCTION

CD28 CD28 molecule Essential for CD4 T cell proliferation

CD40 CD40 molecule Mediates in CD40 responses, inflammation, and other Immune responses

C4B Complement component 4B

Codes for Compliment Factor 4 , provides interaction between antigen and compliment components

CD8 Compliment Component 8D

Present on surface of T lymphocytes, Helps recognize Antigens displayed by Antigen Presenting Cells

CCR6 Chemokine receptor 6

Helps B cell differentiation , and maturation. Expressed by CD4 and CD8 (natural killer cells)

CCR4 Chemokine (C-C motif) receptor 4

Codes for proteins which control development, homeostasis and function of Immune System

CCr7 Chemokine receptor 7

Codes for proteins which activates B and T lymphocytes

CCL2 (C-C motif) ligand

Codes for Cytokine Production which are responsible for inflammatory response

IL2 Interleukin 2 Encodes for protein which releases a cytokine important for B and T cell proliferation

IL10 Interleukin 10 Codes 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; 5:1553-1558

• Deng MC, Eisen HJ, Mehra MR. Noninvasive Discrimination of Rejection in Cardiac Allograft Recipients Using gene Expression Profiling. American Journal of Transplantation 2006; 6:150-160

• Deng MC. Cardiac Transplantation. Heart 2002; 87:177-184

• Michaels PJ, Fishbein MC, Colvin RB. Humoral rejection of Human organ Transplants. Springer Seminars in Immunopathology 2003 119-140.

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