T cell receptor V beta repertoire in an acute infection of rhesus monkeys with simian immunodeficiency viruses and a chimeric simian-human immunodeficiency virus.

Abstract

SummaryChanges in T cell receptor (TCR) V/3 repertoire and their correlation with virologic events were investigated in rhesus monkeys after acute infection with the simian immunodeficiency virus (SIV). 11 genetically defined rhesus monkeys were experimentally infected with SIVm~ or a chimeric simian-human immunodeficiency virus (SHIV), and their peripheral blood lymphocytes (PBL) and lymph nodes were prospectively assessed for TCR V~ gene expression. PBL and lymph nodes of the acutely infected monkeys demonstrated an expansion of selected V~-expressing T lymphocyte subpopulations as early as 3 d after infection. These expanded V/~-expressing lymphocyte subpopulations were comprised predominantly of CD8+ cells. Six of seven infected monkeys sharing a single electrophoreticaUy defined major histocompatibility complex class I allele exhibited a similar expansion of VB14-expressing PBL. Sequence analyses of V-D-J segments of TCR-fl cDNA indicated that the VB-expressing T cell subpopulation expansion can be oligoclonal. SIVm,c-specific CDS+ cytotoxic T lymphocytes were demonstrated in both PBL and lymph nodes of the infected monkeys at the time expansion of the selected V3-expressing cell subpopulations was seen. Finally, the expansion of the selected V3-expressing lymphocytes in PBL coincided with the emergence and clearance of SIV p27 from the plasma of the infected monkeys. These results demonstrate that acute infection of rhesus monkeys with SIVm~ or SHIV results in an expansion of CD8 + lymphocyte subpopulations expressing selected V3 gene families. The selectively expanded T lymphocytes may contribute to early viral clearance after acute SIVmac or SHIV infection.

An Autoantigen-Specific, Highly Restricted T Cell Repertoire Infiltrates the Arthritic Joints of Mice in an HLA-DR1 Humanized Mouse Model of Autoimmune Arthritis

“…CDR3 length analysis was performed by two rounds of nested PCR as described previously (20). The initial TCR-BV gene family-specific PCR was carried out in separate reactions for each BV in 25 ml total volumes consisting of 0.5 ml cDNA, 2.5 ml 103 PCR buffer, 0.5 ml 10 mM 29-deoxynucleoside 59-triphosphates, 1 ml 10 mM TCR-BV forward primer (Table I), 1 ml 10 mM Cb reverse primer (Table I), and 19.5 ml H 2 O.…”

The TCR Repertoire of an Immunodominant CD8+ T Lymphocyte Population

“…The PCR was performed for 30 cycles under the following conditions: 95°C for 30 s, 60°C for 1 min, and 72°C for 2 min. The PCR products were digested with EcoRI and XbaI and ligated into the pSP65 plasmid (Promega, Madison, WI) for cloning and sequencing (2). For frequency analyses, 80 -120 clones were sequenced and analyzed for each cDNA sample.…”

“…The emergence of virus-specific CTL during primary HIV-1 and SIV infection is associated with a reduction in early viremia (1)(2)(3)(4). Moreover, high-frequency virus-specific CTL responses appear to contribute to a decrease in virus load and a delay in disease progression in chronically HIV-1-infected persons (5)(6)(7)(8).…”

Combined megaplex TCR isolation and SMART-based real-time quantitation methods for quantitating antigen-specific T cell clones in mycobacterial infection

“…Calculation and expression of frequencies of antigen-specific Vβ+ T cell clones-To quantitate clonotypic T cell clones in a T cell pool, individual clonotypic TCR transcripts were measured for their frequencies in total TCR β transcripts (Cβ), as previously described (Chen et al, 1995(Chen et al, , 2001. To this end, the threshold cycle (C T ) values for individual Vβ-bearing clones (C T Vβ) and Cβ (C T Cβ) were calculated in each of cDNA samples.…”

“…Over the past decade, we have characterized molecular aspects of MHC-restricted cytotoxic T lymphocytes, superantigen-reactive Vβ + T cell subpopulations, mycobacterium-selected CDR3-bearing T cells, and MHC/viral peptide tetramer-bound T cells in monkey models of infections (Chen et al, 1995(Chen et al, , 1996(Chen et al, , 2001Kou et al, 1998;Zhou et al, 1999). We also utilized the Altas Switch Mechanism At the 5′-end of Reverse Transcript (SMART)-derived cDNA to clone and sequence TCR in a MHC/peptide tetramer-reactive CD8 T cell population (Chen et al, 2001).…”