Increased proteolysis of diphtheria toxin by human monocytes after heat shock: a subsidiary role for heat-shock protein 70 in antigen processing.
The expression of heat-shock proteins (hsp) increases after exposure to various stresses including elevated temperatures, oxidative injury, infection and inflammation. As molecular chaperones, hsp have been shown to participate in antigen processing and presentation, in part through increasing the stability and expression of major histocompatibility complex molecules. Heat shock selectively increases human T-cell responses to processed antigen, but does not affect T-cell proliferation induced by non-processed antigens. Here, we have analysed the mechanisms by which stress such as heat shock, and the ensuing hsp over-expression affect the processing of diphtheria toxin (DT) in peripheral blood monocytes. We found that heat shock increased DT proteolysis in endosomes and lysosomes while the activities of the cathepsins B and D, classically involved in DT proteolysis, were decreased. These effects correlated with the heat-shock-mediated increase in hsp 70 expression observed in endosomes and lysosomes. Actinomycin D or blocking anti-hsp 70 antibodies abolished the heat-shock-mediated increase in DT proteolysis. These data indicate that the increased expression of hsp 70 constitutes a subsidiary mechanism that facilitates antigen proteolysis in stressed cells. Confirming these data, presentation by formaldehyde-fixed cells of DT proteolysates that were obtained with endosomes and lysosomes from heat-shocked peripheral blood monocytes showed higher stimulation of T cells than those generated with endosomes and lysosomes from control peripheral blood monocytes.
Polla
Barbara S
BS
Forever Laser Institut, Geneva, Switzerland. barbara.polla@vtx.ch
Gabert
Françoise
F
Peyrusse
Brigitte M-N
BM
Jacquier-Sarlin
Muriel R
MR
D017951:Antigen Presentation / Q000276:immunology*; D002401:Cathepsin B / Q000378:metabolism; D002402:Cathepsin D / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D004167:Diphtheria Toxin / Q000276:immunology*; D011992:Endosomes / Q000276:immunology; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology*; D018869:Heat-Shock Response / Q000276:immunology*; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D008247:Lysosomes / Q000276:immunology; D009000:Monocytes / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology
D004167:Diphtheria Toxin; D018840:HSP70 Heat-Shock Proteins; D002401:Cathepsin B; D002402:Cathepsin D