The long-term objective of this project is to study the components of the ternary complex of antigen, T cell receptor (TCR) and MHC class I molecule whose interaction leads to T helper cell activation structure/function relationships between components of this ternary complex are currently being analyzed using murine T cell clones specific for well defined epitopes on the model protein antigen sperm whale myoglobin (Spw Mb). A panel of T cell clones has been generated from DBA/2 mice, reactive with Spw mb epitopes characterized with overlapping sets of synthetic peptides.
The specific aims are as follows: 1) To further characterize the antigen and MHC specificity of these clones and of additional mb clones generated in the mouse strains B10.D2 BALB/c and D2.GD. These studies will include the use of horse myoglobin and substituted peptides to identify residues within the epitope important of interaction with the TCR or restricting Class II molecules 2) To sequences alpha and beta chains from matched sets of clones that shave either epitope specificity or MHC restriction in common, and to compare these sequences with those already derived from independent clones that share epitope specificity and MHC restriction. The relationship between TCR and antigen specificity/MHC restriction will be further addressed by transfection of chosen TCR alpha or beta chain genes into Mb- reactive T cell hybridomas expressing endogenous TCR genes of known sequence; 3) To make use of class II (I-Ed) variants generated by site-directed mutagenesis to analyze the interaction of TCR and antigen with the restricting class II molecule. These experiments should determine whether the same set of TCR genes are used to respond to a particular epitope in association with an MHC class II gene product regardless of the non-MHC background if they do, then this result would support the concept that HLA associated diseases might result form the formation of a particular ternary complex involving a shared MHC restriction molecule, a common """"""""disease-associated"""""""" epitope, and a shared T cell receptor.
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