title: Expression of a class II major histocompatibility complex (MHC) heterodimer in a lipid-linked form with enhanced peptide/soluble MHC complex formation at low pH.
abstract: A murine class II major histocompatibility complex (MHC) heterodimer, Ek, expressed as a glycan-phosphatidyl inositol-anchored chimera on Chinese Hamster Ovary cells, can present peptides, but not processed antigen to T cells. This chimeric MHC requires a 100-times higher peptide concentration to achieve a two- to four-times lower level of T cell stimulation. Cleavage with phosphatidylinositol-specific phospholipase C and purification result in large quantities of heterodimer in a water-soluble form. Plates coated with this material and then incubated with peptide can efficiently stimulate the appropriate T cell hybridomas. This stimulation is significantly enhanced when peptides are preincubated with the plate-bound MHC molecules in a pH range (5.0-5.5) similar to that of late endosomes. More than half of the soluble Ek molecules can form a specific complex with cytochrome c peptides in this pH range. This suggests that class II MHC molecules undergo distinct conformational changes in endosomal compartments that render them more capable of forming functional complexes with peptide antigens, irrespective of other cell components.
authors.affiliation: Department of Microbiology and Immunology, Stanford University, California 94305.
authors.forename: D A
authors.initials: DA
authors.lastname: Wettstein
authors.forename: J J
authors.initials: JJ
authors.lastname: Boniface
authors.forename: P A
authors.initials: PA
authors.lastname: Reay
authors.forename: H
authors.initials: H
authors.lastname: Schild
authors.forename: M M
authors.initials: MM
authors.lastname: Davis
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002678:Chimera; D003001:Cloning, Molecular; D005434:Flow Cytometry; D015870:Gene Expression; D005802:Genes, MHC Class II; D006017:Glycolipids / Q000502:physiology; D017261:Glycosylphosphatidylinositols; D000949:Histocompatibility Antigens Class II / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D008285:Major Histocompatibility Complex; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010716:Phosphatidylinositols / Q000502:physiology; D011485:Protein Binding; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection
chemical_list: D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D000949:Histocompatibility Antigens Class II; D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D010716:Phosphatidylinositols