| 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... |
