A mutant antigen-presenting cell defective in antigen presentation expresses class II MHC molecules with an altered conformation.
Ag presentation by APC to class II MHC-restricted T cells involves a sequence of events: 1) intracellular processing of protein Ag into immunogenic peptides, 2) specific binding of peptides to class II MHC molecules, and then 3) transport of the MHC-peptide complexes to the plasma membrane. The critical event in the activation of T cells by APC is the recognition of MHC-associated antigenic determinants by the TCR/CD3 complex. In this report we describe the isolation and characterization of a mutant APC with a defect in an intracellular process that results in its inability to form MHC-peptide complexes for recognition by T cells. The mutant APC cannot present many different protein Ag with both I-A and I-E molecules but is able to present processing-independent peptides. The functional defect in the mutant APC is not caused by either a decrease in expression or a structural mutation in class II MHC molecules. Further, there is no mutation in the invariant chain (li) and it displays a normal kinetics of association and dissociation from the class II MHC molecules during biosynthesis. Although the mutation is not in the genes encoding for the class II MHC molecules or li, the mutant APC expresses class II MHC molecules with distinct serological epitopes suggestive of an altered conformation. Pulse-chase experiments suggest that a conformational difference between I-Ad molecules of wild-type and mutant cells occurs after the class II molecules exit from the endoplasmic reticulum but while they are still associated with li. The mutant cell produces few compact (SDS-resistant) class II heterodimers. This mutant APC provides a tool for studying the cell biology of Ag processing and presentation.
Dang
L H
LH
Division of Lymphocyte Biology, Dana-Farber Cancer Institute, Boston, MA 02115.
Lien
L L
LL
Benacerraf
B
B
Rock
K L
KL
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D000944:Antigens, Differentiation, B-Lymphocyte; D002451:Cell Compartmentation; D002478:Cells, Cultured; D000949:Histocompatibility Antigens Class II / Q000276:immunology* / Q000648:ultrastructure; D006825:Hybridomas / Q000276:immunology; D066298:In Vitro Techniques; D051379:Mice; D008815:Mice, Inbred Strains; D009154:Mutation; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D013601:T-Lymphocytes / Q000276:immunology*
D000944:Antigens, Differentiation, B-Lymphocyte; D000949:Histocompatibility Antigens Class II; C045084:invariant chain