结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

结肠癌富伴侣分子疫苗的制备及其抗癌作用研究

Objective To prepare the anticolon carcinoma vaccine with rich chaperone peptide and to examine its anticancer immunological efficacy. Methods CT26 colon carcinoma cells were cultured in 1 mg/L Trichosanthin 1640 medium at different temperatures to induce the chaperone expression and promote the synthesis of antigen peptides. Groups of these cells treated under the different condition were lysed by the sonic disintegration, and the lysates were centrifuged. The rawpurified proteins were obtained from the supernatants by precipitating with saturated ammonium sulfate and removing the molecules below 50 000 and above 300 000 in molecular weight via dialysis. Furthermore, the proteins with the molecular weights in 70 000, 90 000, 95 000, 110 000 and 170 000 were collected through gel filtration and SDS-PAGE. The purified proteins were analysed by Western blotting, and inspected on the anticancer immunological effects including lymphocyte proliferation and the activities of NK and CTL. Results Major of the chaperone peptides of anticancer effects in CT26 cells, including antigen peptides joining with HSP70, HSP90, gp96, HSP110 and HSP170, was satisfactorily extracted and condensed, and rich chaperone peptide composites were successfully obtained. The composites prepared under various condition could all enhance lymphocyte proliferation and the activities of CTL and NK (P<0.01). Conclusions The rich chaperone peptide composites are successfully prepared via dialysis, salt fractionation and gel filtration combined with SDS-PAGE. Both the heat stress and Trichosanthin can increase the composites, which treated by 42℃ heat stress and Trichosanthin are found to possess the strongest anticancer efficacy.     

肿瘤Ki-67抗原HLA-A2限制性细胞毒性T淋巴细胞表位的鉴定

Objective To identify the HLA-A2-restricted CTL epitopes encoded by antigen segment of Ki-67 for preparation of tumor vaccines. Methods Two epitope prediction databases ( BIMAS and SYFPEITHI) were used to predict the epitopes of Ki-67, and 7 peptides were synthesized as the candi-dates. The prediction was evaluated by peptide-binding test. ELISPOT was used to inspect the IFN-γ secre-ted by CTLs in order to reflect the immunogenicity of the peptides. Results The purities of synthetic pep-tides and positive control peptide were all above 95%. Peptide-binding test displayed that LQGETQLLV (280-288) of Ki-67 could bind with HLA-A2 molecule more strongly. ELISPOT assay displayed that LQGETQLLV could induce activation of specific CTLs. Conclusion LQGETQLLV (280-288) might be the HLA-A2-restricted CTL epitope of Ki-67 antigen.