The document discusses T cell receptor development and evaluation methods. It provides an overview of T cell development including the development of alpha beta and gamma delta T cells from germline genes. It also discusses the rearrangement of variable, diversity, and joining domains in T cell receptor genes. Evaluation methods discussed include CDR3 length analysis using immunoscope technology, clonotypic TCR tracking through peptide stimulation and sorting of T cells, and microarray analysis of beta chain repertoire.
12. CDR regions Stewart-Jones et al 2003 Nature Immunology Diagram highlighting the CDR loops of the T cell clone JM22 in complex with MP(58−66)−HLA-A2. The CDR loops are colored as follows: V α CDR1, green; V α CDR2, red; V α CDR3, blue; V β CDR1, magenta; V β CDR2, orange; V β CDR3, cyan. * MP(58-66) Influensa virus matrix protein
13. CDR3 length analysis Figure 1 . Description of the immunoscope technology. Lymphocytes derived from the blood are stimulated in vitro in the presence of the antigen. 3 days later RNA is extracted and cDNA is synthesized. 24 different PCR amplifications are performed (one for each V /C combination). 12 run off reaction s are performed for each V n-C PCR product, each one using a fluorescent reverse primer specific for one J . Finally each run-off’s products are separated on the basis of length. Ria et al 2001 Current Molecular Medicine