An immunodominant MHC class II-restricted tumor antigen is conformation dependent and binds to the endoplasmic reticulum chaperone, calreticulin.
There is accumulating evidence that CD4(+) T cell responses are important in antitumor immunity. Accordingly, we generated CD4(+) T cells against the murine CT26 colon cancer. Three of three independent CT26-specific CD4(+) hybridomas were found to recognize the high m.w. precursor of the env gene product gp90. The CD4(+) response was completely tumor specific in that the same glycoprotein expressed by other tumors was not recognized by the CT26-specific hybridomas. The recognition of gp90 by the hybridomas was strictly dependent on the conformation of gp90. Different procedures that disrupted the conformation of the glycoprotein, such as disulfide bond reduction and thermal denaturation, completely abrogated recognition of gp90 by all three hybridomas. In CT26 cells, but not in other tumor cells tested, a large proportion of gp90 was retained in the endoplasmic reticulum, mostly bound to the endoplasmic reticulum chaperone, calreticulin. Although calreticulin was not essential for the stimulation of the gp90-specific hybridomas, most of the antigenic form of gp90 was bound to it. The antigenicity of gp90 correlated well with calreticulin binding, reflecting the fact that specificity of binding of calreticulin to its substrate required posttranslational modifications that were also necessary for the generation of this tumor-specific CD4(+) epitope.
Golgher
D
D
Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, Oxford, United Kingdom.
Korangy
F
F
Gao
B
B
Gorski
K
K
Jaffee
E
E
Edidin
M
M
Pardoll
D M
DM
Elliott
T
T
D000818:Animals; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037282:Calreticulin; D004721:Endoplasmic Reticulum / Q000276:immunology / Q000378:metabolism*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology; D005260:Female; D015686:Gene Products, env / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000378:metabolism*; D006358:Hot Temperature; D006825:Hybridomas / Q000378:metabolism; D016056:Immunodominant Epitopes / Q000378:metabolism*; D009052:Leukemia Virus, Murine / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008822:Mice, Transgenic; D011485:Protein Binding / Q000276:immunology; D011487:Protein Conformation; D011489:Protein Denaturation; D012261:Ribonucleoproteins / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000951:Antigens, Neoplasm; D002135:Calcium-Binding Proteins; D037282:Calreticulin; D018984:Epitopes, T-Lymphocyte; D015686:Gene Products, env; D000949:Histocompatibility Antigens Class II; D016056:Immunodominant Epitopes; D012261:Ribonucleoproteins