A new purification method for enhancing the immunogenicity of heat shock protein 70-peptide complexes

Gao,Yanwei; Chen,Xia; Gao,Weishi; Yang,Yong; Ma,Hulin; Ren,Xinjun

doi:10.3892/or.2012.2051

A new purification method for enhancing the immunogenicity of heat shock protein 70-peptide complexes

Authors:
- Yanwei Gao
- Xia Chen
- Weishi Gao
- Yong Yang
- Hulin Ma
- Xinjun Ren
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Affiliations: Department of Oncology, Inner Mongolia People's Hospital, Hohhot 010017, Inner Mongolia, P.R. China, Inner Mongolia Red Cross Blood Center, Hohhot 010017, Inner Mongolia, P.R. China, Tianjin Medical University Eye Center, Tianjin 300384, P.R. China
Published online on: September 21, 2012 https://doi.org/10.3892/or.2012.2051
Pages: 1977-1983
Copyright: © Gao et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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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.

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