Functional Proteomics to Identify Moderators of CD8+ T-Cell Function in Melanoma
Annual rept. 1 Sep 2013-31 Aug 2014
VIRGINIA UNIV CHARLOTTESVILLE RECTOR AND VISTORS OF THE UNIVERSITY OF VIRGINIA
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In the current funding period we have optimized phage-screens to select clones that differentially bind to either tumor infiltrating cytotoxic CD8 lymphocytes, activated CD8 lymphocytes from the spleen, or un-activated naive CD8 T cells. We have developed a high-throughput flow cytometric approach that allows us to screen the specificity of several phage clones for each of these CD8 populations. Using this initial approach we have identified 17 phage that selectively bind TIL rather than effector cells. However, none of these phage influenced CD8 TILl expansion or function in vitro. Using a novel NextGeneration sequencing approach, we have further defined another 1,000,000 phage that selectively bind TIL, of which 100,000 are unique reads. Highly represented phage have been subcloned and are being tested for in vitro function. We have identified one phage that augments T cell expansion in vitro.
- Medicine and Medical Research