Chemical chaperones enhance superantigen and conventional antigen presentation by HLA-DM-deficient as well as HLA-DM-sufficient antigen-presenting cells and enhance IgG2a production in vivo.
Chemical chaperones, first defined in studies of mutant cystic fibrosis transmembrane conductance regulator proteins, are small molecules that act as stabilizers of proteins in their native state and have the ability in some cases to rescue protein-folding mutants within cells. HLA-DM is an MHC II-specific molecular chaperone that facilitates peptide loading onto MHC II proteins and also stabilizes empty MHC II molecules prior to their acquisition of antigenic peptides. APC that lack HLA-DM exhibit quantitative defects in protein Ag as well as superantigen presentation. Here we show that both the superantigen and protein presentation defect in MHC II-transfected, HLA-DM-deficient T2 cells can be partially overcome by treating the APC with the chemical chaperones glycerol, DMSO, or trimethylamine oxide. These chemical chaperones also enhance superantigen and conventional Ag presentation by wild-type APC. In vivo, glycerol was found to act as an adjuvant and resulted in enhanced IgG2a production to trinitrophenyl-keyhole limpet hemocyanin (TNP-KLH). In vitro, the enhancement of Ag presentation by chemical chaperones was found to take place at the level of the APC and took several hours to develop. Subcellular fractionation experiments show that HLA-DM enhances presentation of peptides by dense endosome fractions whereas chemical chaperones enhance presentation by light membrane fractions (early endosome or plasma membrane). The mechanism by which these chemical chaperones augment Ag presentation is not defined, but flow cytometric analysis suggests that the enhancement may be due to a subtle effect on the stability of several different proteins at the cell surface.
Department of Immunology, University of Toronto, Ontario, Canada.
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B
Ghumman
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Bertram
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Watts
D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D000917:Antibody Formation / Q000187:drug effects; D017951:Antigen Presentation / Q000187:drug effects*; D000938:Antigen-Presenting Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D011992:Endosomes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D004768:Enterotoxins / Q000276:immunology / Q000378:metabolism; D005260:Female; D005990:Glycerol / Q000494:pharmacology; D006681:HLA-D Antigens / Q000235:genetics* / Q000276:immunology; D006241:Haptens; D006433:Hemocyanins / Q000008:administration & dosage / Q000276:immunology; D000949:Histocompatibility Antigens Class II; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis*; D008744:Methylamines / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D014176:Protein Biosynthesis; D013211:Staphylococcus aureus / Q000276:immunology; D018089:Superantigens / Q000276:immunology / Q000378:metabolism*
D000276:Adjuvants, Immunologic; D004768:Enterotoxins; C416189:H2-M antigens; D006681:HLA-D Antigens; C084245:HLA-DM antigens; D006241:Haptens; D000949:Histocompatibility Antigens Class II; D007074:Immunoglobulin G; D008744:Methylamines; D018089:Superantigens; C054401:trinitrophenyl keyhole limpet hemocyanin; C026066:enterotoxin A, Staphylococcal; D006433:Hemocyanins; C005855:trimethyloxamine; D005990:Glycerol; D004121:Dimethyl Sulfoxide