Molecular characterization of a subtype of DQw1 recognized by hapten-specific T cells |
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Authors: | Jeffrey H Hanke Richard G Cook Joseph W Leone Mai Van Robert R Rich |
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Institution: | (1) Howard Hughes Medical Institute Laboratory and the Departments of Microbiology and Immunology and Medicine, Baylor College of Medicine, Houston, Texas, USA |
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Abstract: | Previous studies of HLA-restricted antigen recognition by cloned T cells have frequently demonstrated reactivity that did not correlate precisely with the expression of serologically defined HLA specificities. To further explore such discrepancies, we utilized monoclonal antibody (MoAb) blocking, partial NH2-terminal amino acid sequencing, and Southern blot hybridization techniques to analyze the fine specificity of four autologous trinitrophenyl-specific T cell lines restricted to DR2-linked epitopes. MoAb blocking studies demonstrated that two of these lines recognized determinants on DR molecules while the other two recognized determinants on the same molecule that expresses the DQw1 determinant. However, these latter two lines appeared to recognize a DQw1-related determinant found primarily in association with DR2, but not the other DQw1-associated DR alleles, DR1 and DRw6. To ascertain whether these lines were defining a functional split of DQw1, we performed partial NH2-terminal amino acid sequencing of the molecules precipitated with a DQw1-specific MoAb (Genox 3.53) from different stimulator lines. The results showed that these T cell lines recognized a subtype of DQw1 that is in linkage disequilibrium with DR2. Moreover, we identified characteristic restriction fragment length polymorphisms with a DQ
-specific cDNA that correlated with stimulatory capacity for the DQw1-restricted lines. These results demonstrate that: (1) DQ molecules may provide restriction determinants that are incorrectly assigned to DR molecules on stimulator panel analyses; (2) cloned antigen-specific T cell lines recognize polymorphic regions of class II molecules not distinguished by either conventional typing antisera or xenogeneic MoAb; and (3) the DQw1 epitope(s) is located on a heterogeneous group of DQ molecules that differ from each other in the primary sequence of their chains.Abbreviations used in this paper ATCC
American type culture collection; cpm, counts per minute
- DNA
deoxyribonucleic acid
- EBV
Epstein-Barr virus
- FCS
fetal calf serum
- MoAb
monoclonal antibody
- PBMC
peripheral blood mononuclear cells
- % RAgS
percent relative antigen stimulation
- RFLP
restriction fragment length polymorphism
- SDS
sodium dodecyl sulfate
- S-RPMI
supplemented-RPMI
- TCL
T-cell line
- TNP
trinitrophenyl |
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