title: Quantitative analysis of T cell receptor diversity in clinical samples of human peripheral blood.
abstract: The analysis of T cell receptor diversity provides a clinically relevant and sensitive marker of repertoire loss, gain, or skewing. Spectratyping is a broadly utilized technique to measure global TCR diversity by the analysis of the lengths of CDR3 fragments in each Vβ family. However the common use of large numbers of T cells to obtain a global view of TCR Vβ CDR3 diversity has restricted spectratyping analyses when limited T-cell numbers are available in clinical setting, such as following transplant regimens. We here demonstrate that one hundred thousand T cells are sufficient to obtain a robust, highly reproducible measure of the global TCR Vβ repertoire diversity among twenty Vβ families in human peripheral blood. We also show that use of lower cell number results not in a dwindling of observed diversity but rather in non-reproducible patterns in replicate spectratypes. Finally, we report here a simple to use but sensitive method to quantify repertoire divergence in patient samples by comparison to a standard repertoire profile we generated from fifteen normal donors. We provide examples using this method to statistically evaluate the changes in the global TCR Vβ repertoire diversity that may take place during T subset immune reconstitution after hematopoietic stem cell transplantation or after immune modulating therapies.
authors: Experimental Transplantation and Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-1203, United States. memons@mail.nih.gov
authors: Sarfraz A
authors: SA
authors: Memon
authors: Claude
authors: C
authors: Sportès
authors: Francis A
authors: FA
authors: Flomerfelt
authors: Ronald E
authors: RE
authors: Gress
authors: Frances T
authors: FT
authors: Hakim
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000737:chemistry* / Q000276:immunology*; D019411:Clinical Laboratory Techniques / Q000379:methods; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000737:chemistry* / Q000276:immunology*; D015166:Monitoring, Immunologic / Q000379:methods*; D011948:Receptors, Antigen, T-Cell / Q000097:blood* / Q000276:immunology*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology; D015203:Reproducibility of Results
chemical_list: D011948:Receptors, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta