title: Anti-heat Shock 70 kDa Protein Antibody Induced Neuronal Cell Death.
abstract: Heat shock protein 70 (Hsp70) is not only a molecular chaperone in cytosol, but also presents in synaptic plasma membranes. To detect plasmalemmal Hsp70 (pl-Hsp70), neurons were immunostained with anti-Hsp70 antibody without permeabilization and fixation. Dotted immunofluorescent signals at neuronal cell bodies and neurites indicated the localization of Hsp70 on the neuronal cell surface. To target only pl-Hsp70, but not cytosolic Hsp70, the anti-Hsp70 antibody was applied without permeabilization in the primary culture of rat cortical neurons. The antibody induced neuronal cell death in a concentration-dependent manner. The anti-Hsp70 antibody activated ubiquitin-proteasome pathway, but inactivated caspase-3. A lag time was required for the neurotoxicity of anti-Hsp70 antibody. Hydrogen peroxide was increased in the anti-Hsp70 antibody-treated neurons during the lag time. Catalase suppressed the anti-Hsp70 antibody-reduced cell viability via the plausible inhibition of hydrogen peroxide generation. One of down-streams of hydrogen peroxide exposure is activation of the mitogen-activated protein kinase (MAPK) signaling cascade. The neurotoxicity of anti-Hsp70 antibody was partially ascribed to c-Jun N-terminal kinase among MAPKs. In conclusion, the anti-Hsp70 antibody targeted pl-Hsp70 on the neuronal cell surface and induced neuronal cell death without complement. Furthermore, hydrogen peroxide appeared to mediate the neuronal cell death, which was accompanied with the enhancement of the ubiquitin-proteasome pathway and the suppression of caspase in a different fashion from the known cell death.
authors.affiliation: Division of Physiology, Department of Pharmaceutical Health Care, Faculty of Pharmaceutical Sciences,  Himeji Dokkyo University.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Hiromi
authors.initials: H
authors.lastname: Koma
authors.forename: Sayaka
authors.initials: S
authors.lastname: Nishii
authors.forename: Tatsurou
authors.initials: T
authors.lastname: Yagami
mesh_terms: D000818:Animals; D000906:Antibodies / Q000633:toxicity*; D000888:Antibodies, Anti-Idiotypic / Q000633:toxicity; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005260:Female; D006041:Goats; D018840:HSP70 Heat-Shock Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000473:pathology*; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000906:Antibodies; D000888:Antibodies, Anti-Idiotypic; D018840:HSP70 Heat-Shock Proteins; D007074:Immunoglobulin G