T cell receptor gene segment utilization by T lymphocyte clones specific for HLA-DR1 alloantigens.

摘要

Transplantation of incompatible tissues leads to allograft rejection. The response evoked involves two principle components of the immune system, polymorphic MHC molecules and antigen specific receptors expressed on T-cells. The interaction of these molecules is highly specific yet poorly understood. The objectives of this study were to characterize the molecular basis of MHC/T-cell receptor (TcR) interactions and to enhance our understanding of allorecognition. This was accomplished by investigating the relationship between TcR gene utilization and allo-MHC restriction patterns using a panel of HLA-DR1 alloreactive T-lymphocyte clones (TLCs).;Based on proliferative patterns, these alloreactive TLCs have been shown to distinguish the Dw1 and Dw20 subgroups of HLA-DR1. To determine the molecular basis for the Dw1 and Dw20 specificities, HLA-DR sequences were derived from one Dw1 and one Dw20 B-lymphoblastoid cell line (LCL). Two amino acid differences were identified within the first domain of the DR ;To examine TcR gene usage by the DR1-specific TLCs, the one-sided polymerase chain reaction (PCR) technique was developed. In this procedure, mRNA was reverse transcribed with oligo (dT), a series of dGTP residues, termed the homopolymer tail, was added using terminal deoxynucleotidyl transferase, and the tailed-cDNA was amplified using primers complementary to the poly (dG) tail and either the TcR C;Using one-sided PCR technology, the TcR gene segments encoding the TcRs expressed by ten DR1 specific TLCs were analyzed. A heterogeneous usage of variable and junctional gene segments was observed. However, several V