title: A new purification method for enhancing the immunogenicity of heat shock             protein 70-peptide complexes.
abstract: When purified from a tumor, certain heat shock protein 70 (HSP70)-peptide             complexes (PCs) can function as effective vaccines against the tumor from which             the complexes were isolated. The immunogenic mechanisms of HSP70 preparations             imply that tumor-derived HSP70-PCs exhibit antigens associated with antigen-presenting             cells such as dendritic cells (DCs), inducing antigen-specific cytotoxic CD8+             T cells. However, some important membrane-resident tumor-associated peptides,             such as the HER-2/neu (c-erbB2) oncogenic protein, cannot be purified from HSP70             by traditional methods. In the present study, a new approach for the purification             of HSP70-PCs from HER-2-overexpressing breast cancer cells was established. The             detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) was             used to obtain more effectual tumor peptides. The new purified product was named             HSP70-HER-2-PC, and its immunological activities were determined. Traditionally             purified HSP70-PCs (without CHAPS) and recombinant human HSP70-HER-2 protein complexes             (recombined in vitro) were used as controls. These three HSP70-associated tumor             antigenic complex pulsed dendritic cells (DCs) were used to stimulate an antitumor             response. The mature DCs pulsed with HSP70-HER-2-PCs stimulated autologous T cells             to secrete higher levels of type I cytokine compared to the two control groups.             Moreover, DCs pulsed with HSP70-HER-2-PCs induced the most specific CD8+ T cells             that specifically killed the same tumor cells. These findings provide a basis             for new approaches in enhancing HSP70-based immunotherapy for HER-2-associated             or other membrane antigenic peptide-related cancers.
authors.affiliation: Department of Oncology, Inner Mongolia People's Hospital, Hohhot 010017, Inner Mongolia, PR China.
authors.forename: Yanwei
authors.initials: Y
authors.lastname: Gao
authors.forename: Xia
authors.initials: X
authors.lastname: Chen
authors.forename: Weishi
authors.initials: W
authors.lastname: Gao
authors.forename: Yong
authors.initials: Y
authors.lastname: Yang
authors.forename: Hulin
authors.initials: H
authors.lastname: Ma
authors.forename: Xinjun
authors.initials: X
authors.lastname: Ren
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology / Q000378:metabolism; D001943:Breast Neoplasms / Q000378:metabolism*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D019496:Cancer Vaccines / Q000276:immunology; D002458:Cell Fractionation; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism; D003902:Detergents; D005260:Female; D018840:HSP70 Heat-Shock Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D007167:Immunotherapy; D007371:Interferon-gamma / Q000096:biosynthesis; D008213:Lymphocyte Activation; D034622:RNA Interference; D034741:RNA, Small Interfering; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000032:analysis
chemical_list: D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D003902:Detergents; D018840:HSP70 Heat-Shock Proteins; D007371:Interferon-gamma; D034741:RNA, Small Interfering; D018719:Erb-b2 Receptor Tyrosine Kinases; D011994:Recombinant Proteins; C508053:ERBB2 protein, human