Multi-chaperone-peptide-rich mixture from colo-carcinoma cells elicits potent anticancer immunity.
BACKGROUND
Chaperones play an important role in inducing anti-cancer immunity. To explore the probability of using chaperone-peptide-rich complexes extracted from colo-carcinoma cells as anti-cancer vaccine, we extracted and prepared chaperone-peptide-rich complexes from CT26 cells, which were subsequently investigated on anti-cancer efficacy.


METHODS
The crude extracts of the CT26 cells treated with heat and Trichosanthin were precipitated with salt and dialyzed to remove proteins below 50kDa and above 300kDa in molecular weight; the proteins with the molecular weights in 70kDa, 90kDa, 95kDa, 110kDa and 170kDa were collected through gel filtration and SDS-PAGE. After confirmation, the purified proteins were used to determine their effects on lymphocyte proliferation, the activities of NK and CTL, tumor suppression and the tumor-bearing mouse survival.


RESULTS
The majority of the chaperone-peptides of anti-cancer immunity in CT26 cells, including HSP70-antigen peptide, HSP90-antigen peptide, gp96-antigen peptide, HSP-110 antigen peptide, HSP170-antigen peptide, was satisfactorily extracted that the multi-chaperone-peptide-rich mixtures were obtained. All the mixtures prepared could elicit lymphocyte proliferation, enhance the activities of CTL and NK, reinforce the tumor suppression and prolong the mouse survival.


CONCLUSIONS
The multi-chaperone-peptide-rich mixtures could be prepared via dialysis and gel filtration combining with SDS-PAGE. Both the heat stress and Trichosanthin could induce and increase the mixtures, of which that treated by 42 degrees C heat and Trichosanthin was found to possess the strongest anti-cancer efficacy.
Huang
Changxin
C
Hangzhou Normal University School of Medicine, Hangzhou, 310015, China. hcx588@hotmail.com
Zhao
Jiangang
J
Li
Zhaoyang
Z
Li
Da
D
Xia
Dajing
D
Wang
Qingqing
Q
Jin
Hongchuan
H
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D015153:Blotting, Western; D019496:Cancer Vaccines / Q000276:immunology / Q000627:therapeutic use*; D003110:Colonic Neoplasms / Q000276:immunology / Q000628:therapy*; D018882:Heat Stress Disorders / Q000276:immunology; D006358:Hot Temperature; D007694:Killer Cells, Natural / Q000276:immunology; D008213:Lymphocyte Activation; D008223:Lymphoma / Q000276:immunology / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D018832:Molecular Chaperones / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology / Q000627:therapeutic use*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D015978:Trichosanthin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000972:Antineoplastic Agents, Phytogenic; D019496:Cancer Vaccines; D018832:Molecular Chaperones; D010446:Peptide Fragments; D015978:Trichosanthin