Identification of the complete expressed human TCR V gamma repertoire by flow cytometry.
Five of the six expressed human TCR V gamma regions (V gamma 2, 3, 4, 8 and 9) can be detected by available mAb. No mAb with specificity for the remaining V gamma 5 region has been described. In this study, we have characterized mAb 56.3, which was obtained after immunization with V gamma 2/3/4/9-negative thymic gamma delta cells. V gamma transcripts were analyzed by inverse PCR and RT-PCR in 56.3+ cell lines and clones derived from peripheral blood. mAb 56.3 recognized all cells that expressed in-frame V gamma 5 transcripts. In addition, mAb 56.3 recognized some but not all cells expressing V gamma 3, but did not react with a large panel of clones expressing V gamma 2, 4, 8 or 9. In combination with anti-V gamma 2/3/4 mAb 23D12, V gamma 5-expressing cells could be clearly identified as 56.3+23D12(-). In some donors, mAb 56.3 also recognized a small fraction of TCR alpha beta cells. At the clonal level, these cells expressed in-frame V gamma 5-J beta-C beta or V gamma 3-J beta-C beta trans-rearrangements. When mAb 56.3 was combined with V gamma 2/3/4-, V gamma 8- and V gamma 9-specific mAb, all peripheral blood gamma delta T cells were stained. Thus, mAb 56.3 supplements the panel of available TCR V gamma-specific mAb. It is now possible to analyze the complete expressed human V gamma repertoire by flow cytometry.
Hinz
T
T
Department of Immunology, Paul Ehrlich Institute, Langen, Germany.
Wesch
D
D
Halary
F
F
Marx
S
S
Choudhary
A
A
Arden
B
B
Janssen
O
O
Bonneville
M
M
Kabelitz
D
D
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001483:Base Sequence; D003429:Cross Reactions; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D016133:Polymerase Chain Reaction; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment; D016176:T-Lymphocyte Subsets / Q000276:immunology
D000911:Antibodies, Monoclonal; D007135:Immunoglobulin Variable Region; D009841:Oligonucleotides; D016692:Receptors, Antigen, T-Cell, gamma-delta