Synthesis and cell surface display of class II determinants by long-term propagated rat T line cells.
We have investigated the capacity of the encephalitogenic BS rat T cell line bs 83 and its variant clone bs 83.III.C6 to synthesize and express RT1.B-specific class II molecule subsets defined by monoclonal antibodies (mAb) MRC-OX6 and MRC-OX3. Earlier studies had indicated that mAb MRC-OX6 recognizes three distinct molecular species: an immature oligomeric polypeptide chain complex comprised of the polymorphic subunits alpha, beta and the invariant proteins of the gamma group; a biosynthetic intermediate composed of post-translationally modified alpha, beta and gamma chain (denoted p35) and a fully glycosylated alpha, beta two-chain complex derived from the plasma membrane. MRC-OX3 was shown to recognize a serologically distinct alpha, beta two-chain complex that coexists with the MRC-OX6-specific heterodimer at the cell surface. Here we show that premutant bs 83 cells were unable to synthesize class II molecules of either set. In contrast endogeneous synthesis by mutant cells of MRC-OX6-specific molecules was demonstrated. Unlike control spleen cells variant cells failed to synthesize the mature MRC-OX3-reactive class II subset. Instead a three-polypeptide chain complex comprised of the terminally glycosylated subunits alpha, beta and invariant chain p35 was present at the cell surface. This complex appears to represent the preserved biosynthetic intermediate that failed to release invariant chain p35 upon its transit into the plasma membrane. These latter observations support our notion of gamma chain-induced epitope diversification during post-translational maturation of RT1.B-specific class II molecules.
D000818:Animals; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D000939:Epitopes / Q000032:analysis / Q000235:genetics*; D000949:Histocompatibility Antigens Class II / Q000096:biosynthesis* / Q000235:genetics; D007114:Immunization; D007123:Immunoelectrophoresis, Two-Dimensional; D007457:Iodine Radioisotopes; D009154:Mutation; D004676:Myelin Basic Protein / Q000276:immunology; D010641:Phenotype; D011110:Polymorphism, Genetic; D011233:Precipitin Tests; D051381:Rats; D011917:Rats, Inbred Lew / Q000276:immunology*; D011919:Rats, Inbred Strains / Q000276:immunology*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D013997:Time Factors
D000939:Epitopes; D000949:Histocompatibility Antigens Class II; D007457:Iodine Radioisotopes; D004676:Myelin Basic Protein