AIM: To find a novel antigen (Ag) presentation strategy to improve the immune responses induced by dendritic cell (DC)vaccine expressing hepatitis C virus (HCV) core antigen (pcDNA3HCV C-Fc) in Balb/c mice (H-2d).METH...AIM: To find a novel antigen (Ag) presentation strategy to improve the immune responses induced by dendritic cell (DC)vaccine expressing hepatitis C virus (HCV) core antigen (pcDNA3HCV C-Fc) in Balb/c mice (H-2d).METHODS: pcDNA3HCV C-Fc plasmid and eukaryotic expression vector pcDNA3 were injected into mice sc. Immune responses to pcDNA3HCV C-Fc were studied. Meanwhile the effect of pcDNA3HCV C-Fc on anti-translated subcutaneous tumor of SP2/0 cells stably expressing HCV C Ag (SP2/0-HCV C-FC) was also studied. Anti-HCV C in serum was detected by enzyme-linked immunoadsordent assay (ELISA) and HCV specific cytotoxic T lymphocyte (CTL) activity was measured by LDH release assay. After 3 wk of DNA immunization,the cells of SP2/0-HCV C-FC were inoculated into mice subcutaneously and tumor growth was measured every 5 d.The survival rate and living time of mice were also calculated.RESULTS: After 4 wk of DC immunization, the A450 nm values of sera in mice immunized with pcDNA3HCV C-Fc-DC and pcDNA3-DC were 0.56±0.17 and 0.12±0.03 respectively. The antibody titres in mice codeliveried with pcDNA3HCV C-Fc with DC were significantly higher than those of mice injected with pcDNA3-DC. The HCV specific CTL activities in mice coinjected with DC and pcDNA3HCV C-Fc or empty expression vectors were(73.2±3.1) % and (24.4±8.8) %, which were significantly higher than those of mice injected with water.The DC vaccine could evidently inhibit tumor growth, prolong the survival time of mice and improve the survival rate of mice and these effects could be improved by HCV C-Fc (pcDNA3HCV C-Fc) gene codelivered.CONCLUSION: DC vaccine has a strong antigenicity in humoral and cellular immunities, which can be promoted by transduced pcDNA3HCV C-Fc expressing HCV C or Fc.Thus, pcDNA3HCV C-Fc-transduced DCs may be a promising candidate for a CTL-based vaccine against HCV.展开更多
An immunostimulatory factor was identified to be secreted by antigen-pulsed maorophages. This factor was able to induce the generation of antigen specific T helper lymphocytes in vitro as well as in vivo. Further in v...An immunostimulatory factor was identified to be secreted by antigen-pulsed maorophages. This factor was able to induce the generation of antigen specific T helper lymphocytes in vitro as well as in vivo. Further in vitro experiments testing for the genetic restriction of this factor indicated that it is a genetically-restricted antigen specific factor (ASF). The Cunningham plaque assay was used to quantify the generation of T helper lymphocytes by measuring the number of plaque forming cells after sequential incubations of antigen-pulsed maorophages with T lymphocytes, and then spleen cells, and finally the TNP-coated sheep red blood cells.展开更多
Objective:To observe the effects of the recombinant chimeric toxin Dsg3EC 1-2PE40 on T and B lymphocytes isolated from Pemphigus Vulgaris (PV) patients to further study its biological therapeutic function for PV. Meth...Objective:To observe the effects of the recombinant chimeric toxin Dsg3EC 1-2PE40 on T and B lymphocytes isolated from Pemphigus Vulgaris (PV) patients to further study its biological therapeutic function for PV. Methods:Recombinant chimeric toxin Dsg3EC 1-2PE40 was first identified, expressed and purified, and then its effects on T and B lymphocytes of PV patients in vitro were detected and quantified by ELISPOT assay and MTT assay.Results:The purity of the expressed protein Dsg3EC 1-2PE40 was up to 80%. In ELISPOT assay, with Dsg3EC 1-2PE40, the overall number of B cells that produce anti-Dsg3 antibodies among PV patients was only about 60% of the comparable number with Dsg3EC 1-2. The proliferation of T cells of PV patients was inhibited markedly by Dsg3EC 1-2PE40. There was significant difference between the different groups with Dsg3EC 1-2PE40 and Dsg3EC 1-2.Conclusion:The recombinant chimeric toxin Dsg3EC 1-2PE40 decrease the number of B cells that produce anti-Dsg3 antibodies in PV patients and can inhibit or kill T cells of PV patients in vitro.展开更多
In recent years,the in silico epitopes prediction tools have facilitated the progress of vaccines development significantly and many have been applied to predict epitopes in viruses successfully. Herein,a general over...In recent years,the in silico epitopes prediction tools have facilitated the progress of vaccines development significantly and many have been applied to predict epitopes in viruses successfully. Herein,a general overview of different tools currently available,including T cell and B cell epitopes prediction tools,is presented. And the principles of different prediction algorithms are reviewed briefly. Finally,several examples are present to illustrate the application of the prediction tools.展开更多
基金Supported by the National Natural Science Foundation of China, No.30170822
文摘AIM: To find a novel antigen (Ag) presentation strategy to improve the immune responses induced by dendritic cell (DC)vaccine expressing hepatitis C virus (HCV) core antigen (pcDNA3HCV C-Fc) in Balb/c mice (H-2d).METHODS: pcDNA3HCV C-Fc plasmid and eukaryotic expression vector pcDNA3 were injected into mice sc. Immune responses to pcDNA3HCV C-Fc were studied. Meanwhile the effect of pcDNA3HCV C-Fc on anti-translated subcutaneous tumor of SP2/0 cells stably expressing HCV C Ag (SP2/0-HCV C-FC) was also studied. Anti-HCV C in serum was detected by enzyme-linked immunoadsordent assay (ELISA) and HCV specific cytotoxic T lymphocyte (CTL) activity was measured by LDH release assay. After 3 wk of DNA immunization,the cells of SP2/0-HCV C-FC were inoculated into mice subcutaneously and tumor growth was measured every 5 d.The survival rate and living time of mice were also calculated.RESULTS: After 4 wk of DC immunization, the A450 nm values of sera in mice immunized with pcDNA3HCV C-Fc-DC and pcDNA3-DC were 0.56±0.17 and 0.12±0.03 respectively. The antibody titres in mice codeliveried with pcDNA3HCV C-Fc with DC were significantly higher than those of mice injected with pcDNA3-DC. The HCV specific CTL activities in mice coinjected with DC and pcDNA3HCV C-Fc or empty expression vectors were(73.2±3.1) % and (24.4±8.8) %, which were significantly higher than those of mice injected with water.The DC vaccine could evidently inhibit tumor growth, prolong the survival time of mice and improve the survival rate of mice and these effects could be improved by HCV C-Fc (pcDNA3HCV C-Fc) gene codelivered.CONCLUSION: DC vaccine has a strong antigenicity in humoral and cellular immunities, which can be promoted by transduced pcDNA3HCV C-Fc expressing HCV C or Fc.Thus, pcDNA3HCV C-Fc-transduced DCs may be a promising candidate for a CTL-based vaccine against HCV.
文摘An immunostimulatory factor was identified to be secreted by antigen-pulsed maorophages. This factor was able to induce the generation of antigen specific T helper lymphocytes in vitro as well as in vivo. Further in vitro experiments testing for the genetic restriction of this factor indicated that it is a genetically-restricted antigen specific factor (ASF). The Cunningham plaque assay was used to quantify the generation of T helper lymphocytes by measuring the number of plaque forming cells after sequential incubations of antigen-pulsed maorophages with T lymphocytes, and then spleen cells, and finally the TNP-coated sheep red blood cells.
文摘Objective:To observe the effects of the recombinant chimeric toxin Dsg3EC 1-2PE40 on T and B lymphocytes isolated from Pemphigus Vulgaris (PV) patients to further study its biological therapeutic function for PV. Methods:Recombinant chimeric toxin Dsg3EC 1-2PE40 was first identified, expressed and purified, and then its effects on T and B lymphocytes of PV patients in vitro were detected and quantified by ELISPOT assay and MTT assay.Results:The purity of the expressed protein Dsg3EC 1-2PE40 was up to 80%. In ELISPOT assay, with Dsg3EC 1-2PE40, the overall number of B cells that produce anti-Dsg3 antibodies among PV patients was only about 60% of the comparable number with Dsg3EC 1-2. The proliferation of T cells of PV patients was inhibited markedly by Dsg3EC 1-2PE40. There was significant difference between the different groups with Dsg3EC 1-2PE40 and Dsg3EC 1-2.Conclusion:The recombinant chimeric toxin Dsg3EC 1-2PE40 decrease the number of B cells that produce anti-Dsg3 antibodies in PV patients and can inhibit or kill T cells of PV patients in vitro.
基金The National Natural Science Foundations of China (30870131)the National Key Projects in the Infectious Fields (2008ZX10002-011, 2008ZX10004-004)
文摘In recent years,the in silico epitopes prediction tools have facilitated the progress of vaccines development significantly and many have been applied to predict epitopes in viruses successfully. Herein,a general overview of different tools currently available,including T cell and B cell epitopes prediction tools,is presented. And the principles of different prediction algorithms are reviewed briefly. Finally,several examples are present to illustrate the application of the prediction tools.
