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.

1. T cell receptor development and evaluation Anna Merca VIV, SMI Spring 2007

3. Overview of T cell development

4. Development of αβ and γδ T cells

5. Potential germline repertoire Number of functional genes for each domain

6. TRA Rearrangement

7. TRB Rearrangement

8. TRG rearrangement

9. TRD rearrangement

10. TCR complex Abbas & Lichtman 2005

11. TCR complex Abbas & Lichtman 2005

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

15. T-array protocol for Beta Chain Repertoire Bonarius et al 2006

16. Slide representation for T-array Bonarius et al 2006

17. Results for Spectratyping and T-array of Jurkat T cell clone mixed with peripheral blood CD4+ T cells Bonarius et al 2006

18. Sensitivity of various methods fo repertoire analysis Bonarius et al 2006