Introducing endogenous antigens into the major histocompatibility complex (MHC) class II presentation pathway. Both...

The invariant chain (Ii) plays a key role in regulating the antigen presentation function of major histocompatibility complex (MHC) class II molecules. Ii also influences the presentation of usually excluded endogenously synthesized proteins into the MHC class II presentation pathway. To evaluate the role of Ii in the generation of peptide-MHC class II complexes derived from endogenously synthesized proteins, we tested mutant Ii constructs in two model systems. Co-expression of wild-type Ii inhibits the presentation of hen-egg lysozyme (HEL) 35-45/Ak complex, but enhances the presentation of ovalbumin (OVA) 247-265/Ak complex from endogenously synthesized HEL or OVA precursors. The differential sensitivity of these antigens to chloroquine was consistent with their being processed in distinct compartments. Nevertheless, with a panel of Ii deletion constructs we show here that both the Ii-mediated inhibition and enhancement functions require the endosomal targeting and CLIP residues. Surprisingly, the Ii mutant lacking the endoplasmic reticulum lumenal residues 126-215, despite apparently lower expression, was at least as effective as full-length Ii in antigen presentation assays. Thus, alternative pathways exist for processing endogenously expressed antigens, and Ii-mediated inhibition and enhancement of peptide/MHC class II expression depend upon the same regions, with neither requiring the 89 C-terminal, lumenal Ii residues.