Identification of distinct T cell receptor (TCR)-gamma delta heterodimers using an anti-TCR-gamma variable region serum.
T cell hybridomas were generated from CD3+, CD4-, CD8- splenocytes and fetal thymocytes. V gamma 1-expressing proteins present on these murine TCR-gamma delta hybridomas were identified by using an anti-TCR V gamma 1 peptide serum. This antiserum specifically immunoprecipitated 41-kDa TCR V gamma-C gamma 4 chains and 31-kDa TCR V gamma-C gamma 1/2 chains from distinct heterodimers expressed on the TCR-gamma delta T cell hybridomas. The RNA from a hybridoma with a 31-kDa TCR-gamma chain hybridized with a V gamma 1 probe but failed to hybridize with a V gamma 2 probe. In contrast, the RNA from a hybridoma with a 32-kDa TCR-gamma chain hybridized with a V gamma 2 probe. This 32-kDa TCR-gamma chain was not immunoprecipitated by the anti-V gamma 1 serum. These data were consistent with the conclusion that the 31-kDa protein was the product of a V gamma 1 to C gamma 2 rearrangement, whereas the 32-kDa protein was the product of a V gamma 2 to C gamma 1 rearrangement. Furthermore, Southern analyses confirmed that the 32-kDa protein was the product of a V gamma 1.2-J gamma 2 rearrangement, and all three of the 41-kDa TCR-gamma chains were the results of V gamma 1.1-J gamma 4 rearrangements. This was the first demonstration at the clonal level of TCR-gamma proteins which use members of the V gamma 1 gene family, as well as the C gamma 2 constant region. Additional biochemical analyses of the TCR-gamma and -delta proteins from three independently derived C gamma 4-bearing T cell hybridomas suggested that most of the molecular mass diversity observed in the bulk subpopulation of peripheral C gamma 4-containing heterodimers may be contributed by the TCR-delta chains.
Cron
R Q
RQ
University of Chicago, Ben May Institute, IL 60637.
Ezquerra
A
A
Coligan
J E
JE
Houlden
B A
BA
Bluestone
J A
JA
Maloy
W L
WL
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D002236:Carbohydrate Conformation; D006825:Hybridomas / Q000032:analysis / Q000276:immunology; D007106:Immune Sera; D007127:Immunoglobulin Constant Regions / Q000032:analysis; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D008213:Lymphocyte Activation; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping; D011233:Precipitin Tests; D011487:Protein Conformation; D012313:RNA / Q000302:isolation & purification; D011948:Receptors, Antigen, T-Cell / Q000276:immunology / Q000302:isolation & purification*; D013601:T-Lymphocytes / Q000032:analysis / Q000276:immunology
D007106:Immune Sera; D007127:Immunoglobulin Constant Regions; D007135:Immunoglobulin Variable Region; D011948:Receptors, Antigen, T-Cell; D012313:RNA