Simultaneous detection of many T-cell specificities using combinatorial tetramer staining

The direct detection of antigen-specific T cells using tetramers of soluble peptide–major histocompatibilty complex (pMHC) molecules is widely used in both basic and clinical immunology. However, the number of specificities that can be assessed simultaneously has been a major limitation. Here we describe and validate a method using combinations of fluorescent pMHC tetramers to simultaneously detect and enrich for many (≥15) T-cell specificities in a single human blood sample.

[1]  Hidde L. Ploegh,et al.  Discovery of CD8+ T cell epitopes in Chlamydia trachomatis infection through use of caged class I MHC tetramers , 2008, Proceedings of the National Academy of Sciences.

[2]  M. Davis,et al.  Low affinity interaction of peptide-MHC complexes with T cell receptors. , 1991, Science.

[3]  Andrew W. Liu,et al.  Multiplex mapping of CD4 T cell epitopes using class II tetramers. , 2006, Clinical immunology.

[4]  T. Schumacher,et al.  Conditional MHC class I ligands and peptide exchange technology for the human MHC gene products HLA-A1, -A3, -A11, and -B7 , 2008, Proceedings of the National Academy of Sciences.

[5]  T. Schumacher,et al.  Design and use of conditional MHC class I ligands , 2006, Nature Medicine.

[6]  宁北芳,et al.  疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A , 2005 .

[7]  E. Pamer,et al.  Quantitative studies of CD8+ T-cell responses during microbial infection. , 2003, Current opinion in immunology.

[8]  W. Kwok,et al.  HLA Class II-Restricted CD4+ T Cell Responses Directed Against Influenza Viral Antigens Postinfluenza Vaccination 1 , 2003, The Journal of Immunology.

[9]  J. Altman,et al.  A robust method for production of MHC tetramers with small molecule fluorophores. , 2007, Journal of immunological methods.

[10]  Marion Pepper,et al.  Naive CD4(+) T cell frequency varies for different epitopes and predicts repertoire diversity and response magnitude. , 2007, Immunity.

[11]  Philip J. R. Goulder,et al.  Phenotypic Analysis of Antigen-Specific T Lymphocytes , 1996, Science.

[12]  M. Gershwin,et al.  Application of tetramer technology in studies on autoimmune diseases. , 2003, Autoimmunity reviews.

[13]  G. Poste,et al.  Improved flow cytometric analysis of leukocyte subsets: simultaneous identification of five cell subsets using two-color immunofluorescence. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[14]  P. Klenerman,et al.  Ultrasensitive Detection and Phenotyping of CD4+ T Cells with Optimized HLA Class II Tetramer Staining1 , 2005, The Journal of Immunology.

[15]  B. Walker,et al.  Ex vivo analysis of human memory CD4 T cells specific for hepatitis C virus using MHC class II tetramers. , 2003, The Journal of clinical investigation.

[16]  D. Speiser,et al.  Ex vivo analysis of tumor antigen specific CD8+ T cell responses using MHC/peptide tetramers in cancer patients. , 2001, International immunopharmacology.