Cross-presentation of glycoprotein 96-associated antigens on major histocompatibility complex class I molecules requires receptor-mediated endocytosis

J Exp Med. 2000 Jun 5;191(11):1965-74. doi: 10.1084/jem.191.11.1965.


Heat shock proteins (HSPs) like glycoprotein (gp)96 (glucose-regulated protein 94 [grp94]) are able to induce specific cytotoxic T lymphocyte (CTL) responses against cells from which they originate. Here, we demonstrate that for CTL activation by gp96-chaperoned peptides, specific receptor-mediated uptake of gp96 by antigen-presenting cells (APCs) is required. Moreover, we show that in both humans and mice, only professional APCs like dendritic cells (DCs), macrophages, and B cells, but not T cells, are able to bind gp96. The binding is saturable and can be inhibited using unlabeled gp96 molecules. Receptor binding by APCs leads to a rapid internalization of gp96, which colocalizes with endocytosed major histocompatibility complex (MHC) class I and class II molecules in endosomal compartments. Incubation of gp96 molecules isolated from cells expressing an adenovirus type 5 E1B epitope with the DC line D1 results in the activation of E1B-specific CTLs. This CTL activation can be specifically inhibited by the addition of irrelevant gp96 molecules not associated with E1B peptides. Our results demonstrate that only receptor-mediated endocytosis of gp96 molecules leads to MHC class I-restricted re-presentation of gp96-associated peptides and CTL activation; non-receptor-mediated, nonspecific endocytosis is not able to do so. Thus, we provide evidence on the mechanisms by which gp96 is participating in the cross-presentation of antigens from cellular origin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenovirus E1B Proteins / immunology
  • Animals
  • Antigen Presentation / immunology*
  • Antigen-Presenting Cells / immunology
  • B-Lymphocytes / immunology
  • Dendritic Cells / immunology
  • Endocytosis / immunology*
  • H-2 Antigens / immunology
  • HSP70 Heat-Shock Proteins / immunology*
  • Histocompatibility Antigens Class I / immunology*
  • Histocompatibility Antigens Class II / immunology
  • Humans
  • Macrophages / immunology
  • Membrane Proteins / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Molecular Chaperones / immunology*
  • Receptors, Cell Surface / immunology*
  • Tumor Cells, Cultured


  • Adenovirus E1B Proteins
  • H-2 Antigens
  • H-2Kb protein, mouse
  • HSP70 Heat-Shock Proteins
  • Histocompatibility Antigens Class I
  • Histocompatibility Antigens Class II
  • Membrane Proteins
  • Molecular Chaperones
  • Receptors, Cell Surface
  • glucose-regulated proteins