Localization of peptide/MHC class II complexes in macrophages following antigen processing of viable Streptococcus pyogenes

Abstract

The subcellular localization of peptide/MHC complexes was investigated during processing of the surface M5 protein from Streptococcus pyogenes. Bone marrow-derived macrophages were pulsed with viable S. pyogenes for 20 min followed by various periods of chase. T hybridoma cells detected complexes of one epitope, M5(17-31) with E(d) on the surface of macrophages within 30 min of chase. In contrast, complexes with another epitope, M5(308-319) with A(d) peaked later. Intracellular localization of peptide/MHC-II complexes was studied by subcellular fractionation and detection of complexes in fractions by T hybridoma cells. M5(17-31)/E(d) complexes were detected in light membrane fractions containing plasma membrane and early endosomes by 10-30 min. M5(308-319)/A(d) complexes were detected in these light membranes after 3 h of chase. Thus, the time course of M5(308-319)/A(d) presentation was delayed relative to M5(17-31)/E(d). However, neither type of complex was detected at any time in fractions containing phagosomes. Both species of peptide/MHC complexes localized to endocytic compartments, indicating a role for endosomes in presentation of antigens from phagocytosed bacteria.