Project Number: 3635-31320-008-03
Project Type: Reimbursable

Start Date: Jun 01, 2004
End Date: May 30, 2008

Objective:
The specific aims of this project are: 1). Compare immune responses following Marek's disease virus (MDV) infection between susceptible and resistant lines of chickens. 2). Compare cytotoxic T lymphocyte (CTL) responses between strong and weak responders to avian leucosis virus (ALV) infection. 3). Integrate DNA microarray and protein results with whole genome sequencing.

Approach:
Three aims are defined. Aim #1 compares the response of chicken cells to MDV infection. As described previously, we have shown gene expression changes in response to MDV infection in CEFs and between lymphocytes from MD resistant and susceptible chicken lines. We extend these studies by examining the cellular response in MDV-infected cells using laser capture micodissection (LCM) and comparing them to the surrounding uninfected cells. Taking advantage of our characterized inbred lines, we examine how genetic resistance status influences the response. Finally, in addition to profiling gene expression, we will isolate and characterize specific MDV-chicken protein complexes that include host proteins known to be important for the immune response and genetic resistance to MD. Incorporation of defined MDV mutants examines how perturbing key MDV-chicken interactions influence the composition of these functional complexes. Aim #2 is similar to Aim #1 in that we will utilize DNA microarrays to profile gene expression changes in the immune response. In this Aim, ALV subgroups A and J will be the infectious agents, which are known to produce strong or weak CTL responses depending on the MHC haplotype of the chicken. Revealing specific responses in CD4+ or CD8+ lymphocytes as a function of ALV subgroup-chicken MHC combination will provide basic knowledge on what genes are necessary or sufficient to elicit a strong CTL response against ALV. Aim #3 integrates and extends our results using bioinformatics. Comparisons will be made between gene profiling results and, for Aim #1 only, between proteins and their corresponding RNAs. The chicken genome sequence will be integrated by identifying positional candidate genes that confer genetic resistance to MD, and revealing common regulatory elements for key genes. BSL-2; recertified 8/11/06 valid through 8/10/09.