OBJECTIVE: To study the efficacy and explore the mechanism of the anti-tumor immunity elicited by heat shock protein 70-peptide complexes (HSP70-PC) derived from tumor cells. METHODS: Cells culture, flow cytometric an...OBJECTIVE: To study the efficacy and explore the mechanism of the anti-tumor immunity elicited by heat shock protein 70-peptide complexes (HSP70-PC) derived from tumor cells. METHODS: Cells culture, flow cytometric analysis, affinity chromatography for protein purification, SDS-PAGE, Western-blotting and animal experiment were used. RESULTS: HSP70-PC immunization rendered protective effect to both naive tumorl-bearing mice. All of the naive mice obtained complete resistance to Hcaf cell attack; 40% of the tumor-bearing mice survived for over 90 days, whereas the mice of control group died within 2 weeks (P展开更多
Objective The aim of this study was to enhance the treatment effect of tumor purified autogenous heat shock protein 70-peptide complexes(HSP70-PCs)on HER-3-overexpressing breast cancer.Methods In this study,we first s...Objective The aim of this study was to enhance the treatment effect of tumor purified autogenous heat shock protein 70-peptide complexes(HSP70-PCs)on HER-3-overexpressing breast cancer.Methods In this study,we first studied the expression of HER-3 in breast cancer tissues and its relationship with patient characteristics.We then purified HSP70-PCs from primary breast cancer cells with different HER-2 and HER-3 expression profiles and determined the cytotoxicity of autogenous dendritic cells(DCs)and CD8+T cells induced by these complexes.Third,recombinant human HSP70-HER-3 protein complexes were used to inhibit the autogenous HSP70-PCs purified from HER-3-overexpressing breast cancer cells,and the resulting immunological response was examined.Results The results show that HSP70-PCs can be combined with recombinant HSP70-HER-3 protein complexes to induce stronger immunological responses than autogenous HSP70-PCs alone and that these treatments induce autogenous CD8+T cell killing of HER-3-positive breast cancer cells.Conclusion These findings provide a new direction for HSP70-DC-based immunotherapy for patients with HER-3-overexpressing breast cancer.展开更多
Objective The aim of this study was to purify effective tumor peptide complexes from human melanoma cell lines to enhance the treatment effects on melanoma.Methods We purified heat shock protein 70(HSP70)-peptide comp...Objective The aim of this study was to purify effective tumor peptide complexes from human melanoma cell lines to enhance the treatment effects on melanoma.Methods We purified heat shock protein 70(HSP70)-peptide complexes(PCs)from human melanoma cell lines A375,A875,M21,M14,WM-35,and SK-HEL-1.We named the purified product as M-HSP70-PCs and determined its immunological activities.Autologous HSP70-PCs purified from primary tumor cells of melanoma patients(9 cases)were used as controls.These two tumor antigenic complexes were loaded into dendritic cells(DCs)and used to stimulate an antitumor response against tumor cells in the corresponding patients.Results Mature DCs pulsed with M-HSP70-PCs stimulated autologous T cells to secrete the same levels of type I cytokines as the autologous HSP70-PCs.Moreover,DCs pulsed with M-HSP70-PCs endued CIK cells with an equal ability as autologous HSP70-PCs to kill melanoma cells in the patients.Conclusion M-HSP70-PCs may be used as an efficient and generalized tumor antigen in the treatment of DC-based malignant melanoma.展开更多
The preparation of recombinant human HSP70 and its presenting antigen function were investigated. Cultured in glucose free M9ZB medium and induced with IPTG and lactose at a final concentration of 0.02 mmol/L and 5 ...The preparation of recombinant human HSP70 and its presenting antigen function were investigated. Cultured in glucose free M9ZB medium and induced with IPTG and lactose at a final concentration of 0.02 mmol/L and 5 mmol/L respectively, the engineered bacteria carrying expression vector of human HSP70 gene expressed rHSP70 at an efficiency of 60 %. After the purification with DEAE ion exchange chromatography, HSP70 with a purity of higher than 90 % was obtained. The purified product could bind tumor antigen peptide in vitro , and the binding was identified by native PAGE containing 5 % glycerol. HSP70 peptide complex could activate lymphocytes to produce specific cytotoxicity to tumor cells, suggesting that the recombinant human HSP70 could be used as an antigen presenting reagent in tumor therapy.展开更多
目的:探讨热休克蛋白-抗原肽复合物(HSP)疫苗预防肝癌术后复发的效果.方法:回顾性分析32例应用HSP疫苗的原发性肝癌患者,并检测治疗前后T细胞亚群、NK细胞活性和血清IL-2受体水平的变化,计算3 a内的生存率,并与对照组比较.结果:治疗组术...目的:探讨热休克蛋白-抗原肽复合物(HSP)疫苗预防肝癌术后复发的效果.方法:回顾性分析32例应用HSP疫苗的原发性肝癌患者,并检测治疗前后T细胞亚群、NK细胞活性和血清IL-2受体水平的变化,计算3 a内的生存率,并与对照组比较.结果:治疗组术后3 mo CD3+,CD8+,CD4+/CD8+水平与术前和对照组比较有显著性差异(P<0.05),NK细胞活性较治疗前明显升高(P<0.01);治疗组3a生存率为68.8%,与对照组37.5%比较明显升高(P<0.05).结论:肝癌术后应用HSP疫苗能明显提高患者免疫功能,且可预防术后复发、提高远期生存率.展开更多
文摘OBJECTIVE: To study the efficacy and explore the mechanism of the anti-tumor immunity elicited by heat shock protein 70-peptide complexes (HSP70-PC) derived from tumor cells. METHODS: Cells culture, flow cytometric analysis, affinity chromatography for protein purification, SDS-PAGE, Western-blotting and animal experiment were used. RESULTS: HSP70-PC immunization rendered protective effect to both naive tumorl-bearing mice. All of the naive mice obtained complete resistance to Hcaf cell attack; 40% of the tumor-bearing mice survived for over 90 days, whereas the mice of control group died within 2 weeks (P
基金Supported by a grant from the National Natural Science Foundation of China(No.81260392).
文摘Objective The aim of this study was to enhance the treatment effect of tumor purified autogenous heat shock protein 70-peptide complexes(HSP70-PCs)on HER-3-overexpressing breast cancer.Methods In this study,we first studied the expression of HER-3 in breast cancer tissues and its relationship with patient characteristics.We then purified HSP70-PCs from primary breast cancer cells with different HER-2 and HER-3 expression profiles and determined the cytotoxicity of autogenous dendritic cells(DCs)and CD8+T cells induced by these complexes.Third,recombinant human HSP70-HER-3 protein complexes were used to inhibit the autogenous HSP70-PCs purified from HER-3-overexpressing breast cancer cells,and the resulting immunological response was examined.Results The results show that HSP70-PCs can be combined with recombinant HSP70-HER-3 protein complexes to induce stronger immunological responses than autogenous HSP70-PCs alone and that these treatments induce autogenous CD8+T cell killing of HER-3-positive breast cancer cells.Conclusion These findings provide a new direction for HSP70-DC-based immunotherapy for patients with HER-3-overexpressing breast cancer.
基金Supported by a grant from the National Natural Sciences Foundation of Inner Mongolia(No.2017MS(LH)0847)。
文摘Objective The aim of this study was to purify effective tumor peptide complexes from human melanoma cell lines to enhance the treatment effects on melanoma.Methods We purified heat shock protein 70(HSP70)-peptide complexes(PCs)from human melanoma cell lines A375,A875,M21,M14,WM-35,and SK-HEL-1.We named the purified product as M-HSP70-PCs and determined its immunological activities.Autologous HSP70-PCs purified from primary tumor cells of melanoma patients(9 cases)were used as controls.These two tumor antigenic complexes were loaded into dendritic cells(DCs)and used to stimulate an antitumor response against tumor cells in the corresponding patients.Results Mature DCs pulsed with M-HSP70-PCs stimulated autologous T cells to secrete the same levels of type I cytokines as the autologous HSP70-PCs.Moreover,DCs pulsed with M-HSP70-PCs endued CIK cells with an equal ability as autologous HSP70-PCs to kill melanoma cells in the patients.Conclusion M-HSP70-PCs may be used as an efficient and generalized tumor antigen in the treatment of DC-based malignant melanoma.
基金This project was supported by a grant from the NationalNatural Science Foundation of China (Serial No. 39970 32 2 ) and Trans- Century Training Program Foundation for Talentunder the supervision of the Ministry of Education of China
文摘The preparation of recombinant human HSP70 and its presenting antigen function were investigated. Cultured in glucose free M9ZB medium and induced with IPTG and lactose at a final concentration of 0.02 mmol/L and 5 mmol/L respectively, the engineered bacteria carrying expression vector of human HSP70 gene expressed rHSP70 at an efficiency of 60 %. After the purification with DEAE ion exchange chromatography, HSP70 with a purity of higher than 90 % was obtained. The purified product could bind tumor antigen peptide in vitro , and the binding was identified by native PAGE containing 5 % glycerol. HSP70 peptide complex could activate lymphocytes to produce specific cytotoxicity to tumor cells, suggesting that the recombinant human HSP70 could be used as an antigen presenting reagent in tumor therapy.
文摘目的:探讨热休克蛋白-抗原肽复合物(HSP)疫苗预防肝癌术后复发的效果.方法:回顾性分析32例应用HSP疫苗的原发性肝癌患者,并检测治疗前后T细胞亚群、NK细胞活性和血清IL-2受体水平的变化,计算3 a内的生存率,并与对照组比较.结果:治疗组术后3 mo CD3+,CD8+,CD4+/CD8+水平与术前和对照组比较有显著性差异(P<0.05),NK细胞活性较治疗前明显升高(P<0.01);治疗组3a生存率为68.8%,与对照组37.5%比较明显升高(P<0.05).结论:肝癌术后应用HSP疫苗能明显提高患者免疫功能,且可预防术后复发、提高远期生存率.
