Date of Award

2007

Document Type

Thesis

Department

Biological Sciences

Abstract

CD4+ T lymphocytes play a vital role in immune system function but little is known about their role in controlling SV40 T ag-induced tumors in C57BL/6 mice. In order to better investigate their role in the control of tumors, it is necessary to identify CD4 epitope(s) within the Tag. Four CDS epitopes have been identified but the CD4 epitopes have not been identified. To facilitate the identification of the epitopes, T cell hybridomas containing a Lacz reporter gene were used. CD4+ hybridoma clones capable of recognizing SV 40 Large Tumor Antigen (T ag) were obtained from heterogeneous populations through serial dilution and screening with a colorimetric (CPRG) assay in which bone marrow-derived dendritic cells were pulsed with purified SV 40 T ag protein. To map the location of the epitope target of each clone, four of the clones were chosen for use in a screening assay utilizing a library of overlapping synthetic 15mer peptides representing the entire SV 40 T ag amino acid sequence. The identification of such CD4+ epitopes within the T ag will make it possible to determine the immunogenic and regulatory properties of each epitope (in vivo) and determine whether differences in reactivity demonstrated by the various hybridoma clones against the purified SV 40 T ag protein are epitope or hybrid dependent.

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