Examination of the highly diverse CD4 + T-cell repertoire directed against an influenza peptide: a step towards TCR proteomics

Abstract

We combined several recent technological advances in immunology and molecular biology to identify and sequence a large number of T-cell receptor (TCR) genes specific for a particular antigen. We utilized class II MHC tetramers and interferon-gamma surface capture to isolate from samples of peripheral blood the population of CD4(+) T cells responding to a peptide derived from influenza hemagglutinin and restricted by HLA-DR1. Detailed analysis of hundreds of clones from three different patients revealed an extremely diverse repertoire, with little overlap between patients. We observed no dominant usage of particular Vbeta segments nor any clear CDR3 sequence motif in the responding T cells, but most of the clones appear to utilize acidic residues in the CDR1 and CDR3 regions, presumably to interact with the exposed basic residues in the MHC-peptide complex. This methodology could be expanded to a large scale to identify the generalized rules governing TCR-MHC engagement and factors which shape the T-cell repertoire after vaccination and in autoimmune pathologies.

HLA Class II-Restricted CD4+ T Cell Responses Directed Against Influenza Viral Antigens Postinfluenza Vaccination

“…Previous reports have indicated a diversity of V␤ usage among DR0101-restricted HA 306 -318 T cell clones or lines (28,29). The current data provided a direct ex vivo analysis of the V␤ usage of DR0401 HA 306 -318 peripheral T cells.…”

Precursor Frequency, Nonlinear Proliferation, and Functional Maturation of Virus-Specific CD4+ T Cells

“…LCMV-naive gp [61][62][63][64][65][66][67][68][69][70][71][72][73][74][75][76][77][78][79][80] -specific T cells is ϳ100, similar in magnitude to that reported for naive gp [33][34][35][36][37][38][39][40][41] -specific CD8 T cells (5). Thus, the ϳ10-to 50-fold difference in abundance of CD4 ϩ and CD8 ϩ T cells that is present during the anti-LCMV response (29,30) is unlikely to be attributable to a difference in frequency of LCMV-naive precursors.…”

“…In some cases they were backcrossed to the B6.PL background to produce Thy1.1-positive cells. P14 mice express the TCR for the LCMV gp 33 MHC class I epitope (10). Mice were infected by i.p.…”

“…The dotted lines indicate the peptide concentrations required to drive a half-maximal response. D, P14 cells (CD8 ϩ T cells carrying a transgenic receptor for the LCMV MHC class I epitope gp 33 ) and SMARTA cells were adoptively transferred into different C57BL/6 mice, and, 8 days after LCMV infection, the proportion of each population that made IFN-␥ in response to cognate peptide stimulation was determined by ICCS. The p value was calculated using the Student t test.…”

Single-Cell Analysis of T-Cell Receptor Repertoire of HTLV-1 Tax-Specific Cytotoxic T Cells in Allogeneic Transplant Recipients with Adult T-Cell Leukemia/Lymphoma

“…Some recent studies on viral antigen-specific human T cells (42,43) or CTL clones in patients with large granular lymphocyte leukemia (44) and paroxysmal nocturnal hemoglobinuria (45) showed the existence of TCR-β CDR3 amino acid sequences that were completely shared by T cells between unrelated individuals. A previous study on the TCR repertoire of HLA-A*0201-restricted Tax [11][12][13][14][15][16][17][18][19] -specific CTLs also showed the existence of a TCR-β CDR3 amino acid sequence that was completely shared between unrelated HAM/TSP patients, even though limited numbers of patients and T cell clones were analyzed (21).…”