Alternative mechanism by which IFN-gamma enhances tumor recognition: active release of heat shock protein 72.
IFN-gamma exhibits differential effects depending on the target and can induce cellular activation and enhance survival or mediate cell death via activation of apoptotic pathways. In this study, we demonstrate an alternative mechanism by which IFN-gamma enhances tumor recognition, mediated by the active release of Hsp72. We demonstrate that stimulation of 4T1 breast adenocarcinoma cells and K562 erythroleukemic cells with IFN-gamma triggers the cellular stress response, which results in the enhanced expression of total Hsp72 expression without a significant increase in cell death. Intracellular expression of Hsp72 was abrogated in cells stably transfected with a mutant hsf-1 gene. IFN-gamma-induced Hsp72 expression correlated with enhanced surface expression and consequent release of Hsp72 into the culture medium. Pretreatment of tumors with compounds known to the block the classical protein transport pathway, including monensin, brefeldin A, tunicamycin, and thapsigargin, did not significantly block Hsp72 release. However, pretreatment with intracellular calcium chelator BAPTA-AM or disruption of lipid rafts using methyl beta-cyclodextrin completely abrogated IFN-gamma-induced Hsp72 release. Biochemical characterization revealed that Hsp72 is released within exosomes and has the ability to up-regulate CD83 expression and stimulate IL-12 release by naive dendritic cells. Pretreatment with neutralizing mAb or depletion of Hsp72 completely abrogated its chaperokine function. Taken together, these findings are indicative of an additional previously unknown mechanism by which IFN-gamma promotes tumor surveillance and furthers our understanding of the central role of extracellular Hsp72 as an endogenous adjuvant and danger signal.
Bausero
Maria A
MA
Center for Molecular Stress Response, Boston University Medical Center and Boston University School of Medicine, Boston, MA 02118, USA.
Gastpar
Robert
R
Multhoff
Gabriele
G
Asea
Alexzander
A
D015703:Antigens, CD / Q000032:analysis; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000502:physiology*; D006358:Hot Temperature; D006801:Humans; D007136:Immunoglobulins / Q000032:analysis; D007371:Interferon-gamma / Q000494:pharmacology*; D016753:Interleukin-10 / Q000494:pharmacology; D018664:Interleukin-12 / Q000096:biosynthesis; D020014:K562 Cells; D008562:Membrane Glycoproteins / Q000032:analysis; D009369:Neoplasms / Q000276:immunology*; D010641:Phenotype; D000097802:CD83 Antigen
D015703:Antigens, CD; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D007136:Immunoglobulins; D007371:Interferon-gamma; D016753:Interleukin-10; D018664:Interleukin-12; D008562:Membrane Glycoproteins; D000097802:CD83 Antigen