title: [Alterations in heat shock protein 70 kDa levels in human neutrophils under the heat shock conditions].
abstract: Intracellular content of heat shock proteins of 70 kDa family (HSP70) possessing chaperone and cytoprotective functions depends on specialization and functional activity of the cells. The aim of this study was to analyze the dynamics of constitutive and inducible HSP70 levels evoked by heat shock in human neutrophils, the short-lived fraction of white blood cells providing non-specific defense against bacterial pathogens. Biphasic dynamics of the intracellular HSP70 level with an increase and following decrease in 15-30 min after the heat shock was revealed by flow cytometry. This dynamics was similar for constitutive and inducible forms of HSP70. Pre-incubation of neutrophils with cycloheximide, the inhibitor of protein synthesis, did not change the intracellular HSP70 dynamics registered by flow cytometry indicating that the increased HSP70 level detected immediately after the heat shock was not mediated by de novo protein synthesis. It was confirmed by Western blotting analysis of HSP70 intracellular content. It was suggested that the elevated HSP70 level was related to conformational HSP70 molecule changes and to increased availability of HSP70 epitopes for antibody binding. Using a panel of antibodies specific to the N-terminal ATP-binding or C-terminal substrate-binding domains of HSP70 it has been demonstrated by cell immunofluorescence and flow cytometry methods that the heat shock-associated increase of the intracellular HSP70 level was mediated by HSP70 interaction with antibodies recognizing HSP70 substrate-binding domain. It was demonstrated that the decrease of intracellular HSP70 level after heat treatment could be connected with a release of both inducible and constitutive HSP70 into extracellular space. Our data suggest that stress-induced release of HSP70 from neutrophils is regulated by ABC-transporters.
authors.forename: A A
authors.initials: AA
authors.lastname: Boĭko
authors.forename: S S
authors.initials: SS
authors.lastname: Vetchinin
authors.forename: A M
authors.initials: AM
authors.lastname: Sapozhnikov
authors.forename: E I
authors.initials: EI
authors.lastname: Kovalenko
mesh_terms: D003593:Cytoplasm / Q000737:chemistry; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis* / Q000737:chemistry; D018869:Heat-Shock Response; D006358:Hot Temperature; D006801:Humans; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D009504:Neutrophils / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation
chemical_list: D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones