THE INVOLVEMENT OF CD4 T-CELLS IN PROTECTION AGAINST AVIAN HEPATITIS E VIRUS (HEV)

Hepatitis E virus (HEV), a small single strand RNA virus, is the causative agent for Hepatitis E. Hepatitis E is an emerging disease that has affected more than 2 billion people worldwide. Strong HEV specific T-cell responses have been reported in HEV exposed healthy individuals but not in patients with chronic hepatitis E. This suggests that lack of CD4+ or CD8+ or both T-cells may predispose patients to chronic HEV infection in immune suppressed individuals. In order to determine if the T-cell response is important in controlling HEV infection and progression into chronicity, CD4 or CD8 antibodies will be used to deplete chickens of CD4 or CD8 T-cells. The main purpose of this study is to produce an ELISA test to quantify the antibody concentration of CD4 in ascites fluid. Focusing on the C terminus, 5 different truncated CD4 genes were made: CD4 30-428, CD4 219-345, CD4 236-377, CD4 236-487, and CD4 219-428. The genes were amplified, cloned in the pRSET B vector, and transformed into One Shot TOP10 E. coli strain. After restriction endonuclease digestion and DNA sequencing, the clone CD4 236-377 contained CD4 but in the wrong orientation. In future studies, clones containing the CD4 truncated genes will be expressed and used as an antigen in an ELISA test.