Acquisition of peptides by MHC class II polypeptides in the absence of the invariant chain - PubMed (original) (raw)

. 1995 Feb 1;154(3):1048-56.

Affiliations

PMID: 7822782

Acquisition of peptides by MHC class II polypeptides in the absence of the invariant chain

C Hitzel et al. J Immunol. 1995.

Abstract

Association of invariant chain with class II molecules has been suggested to inhibit binding of peptides that are available while the class II complex is present in the endoplasmic reticulum (ER) and subsequently transported to endosomes. We tested HLA-DR-transfected rat2 fibroblast cells lacking expression of invariant chain for their ability to form SDS stable class II dimers indicative of peptide binding in the class II cavity. No SDS-resistant class II dimers originating from short pulse-labeled immunoprecipitates can be identified. Prolonged ER retention of DR polypeptides by Brefeldin A treatment does not induce any stable class II dimers. In pulse-chase experiments, heat labile class II dimers are readily detectable after a 60-min chase, increasing in amounts by 4 h of chase. In vitro incubation of rat2DR cell lysate with DR3-binding peptides converts pulse-labeled class II molecules to SDS-resistant dimers. This indicates the ability of ER-resident DR dimers to bind peptides. Inhibition studies were conducted to define the intracellular site where stable class II complexes are formed in rat2DR cells. The lysosomotropic reagent chloroquine abrogates the appearance of SDS-resistant DR complexes in invariant chain-free rat2DR cells, which is consistent with the impact of chloroquine on peptide loading in other APCs. Leupeptin treatment strongly reduces the amount of heat-labile class II molecules but does not impair peptide loading when DR3-specific peptides were added to viable cells or to cell lysates. This result suggests that leupeptin inhibits intracellular degradation of polypeptides and thereby depletes the endocytic pathway of peptides available for class II binding.

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Acquisition of peptides by MHC class II polypeptides in the absence of the invariant chain - PubMed (original) (raw)