The application of real-time PCR to the analysis of T cell repertoires

The diversity of T-cell populations is determined by the spectrum of antigen-specific T-cell receptors (TCRs) that are heterodimers of α and β subunits encoded by rearranged combinations of variable (AV and BV), joining (AJ and BJ), and constant region genes (AC and BC). We have developed a novel approach for analysis of β transcript diversity in mice with a real-time PCR-based method that uses a matrix of BV- and BJ-specific primers to amplify 240 distinct BV–BJ combinations. Defined endpoints (Ct values) and dissociation curves are generated for each BV–BJ combination and the Ct values are consolidated in a matrix that characterizes the β transcript diversity of each RNA sample. Relative diversities of BV–BJ combinations in individual RNA samples are further described by estimates of scaled entropy. A skin allograft system was used to demonstrate that dissection of repertoires into 240 BV–BJ combinations increases efficiency of identifying and sequencing β transcripts that are overrepresented at inflammatory sites. These BV–BJ matrices should generate greater investigation in laboratory and clinical settings due to increased throughput, resolution and identification of overrepresented TCR transcripts

Dr. Joseph Goldberger and Pellagra : His Life and Legacy

Laupus Library History Collections & the Department of Bioethics & Interdisciplinary Studies sponsor the History of Medicine Presentations as an educational service for the East Carolina University community. The Library hopes that the speakers and topics selected will promote a greater understanding of the historical and philosophical underpinnings of today's health care disciplines.Laupus Library History Collections & the Department of Bioethics & Interdisciplinary Studie