lnterleukin-21(IL-21)is a recently characterized T cell-derived cytokine with a significant homology to IL-2, IL-4 and IL-15.To determine whether IL-21 has broad immunoregulatory activity and can stimulate durable ant...lnterleukin-21(IL-21)is a recently characterized T cell-derived cytokine with a significant homology to IL-2, IL-4 and IL-15.To determine whether IL-21 has broad immunoregulatory activity and can stimulate durable antitumour responses,we constructed mouse IL-21(mIL-21) recombinant plasmid and evaluated its antitumor efficacy.Mouse IL-21 cDNA was amplified from Con A-activated mouse T cells by RT-PCR.Recombinant pcDNA3.1/mIL-21 was constructed and transfected into Sp2/0 cells.Mouse IL-21 expression was analyzed by Western blotting and its activities were detected by ~3H-TdR incorporation and MTT assay.The recombinant pcDNA3.1/mIL-21 was injected s.c.into tumor lump.Tumor size,weight,the activities of CTLs,NK cells and LAK cells and serum IFN-γ,level were measured for evaluating mIL-21 mediated antitumor responses.The results indicated that mIL-21 was correctly expressed in Sp2/0 cells and it can improve the proliferation of T cells and B cells,and enhance NK cytotoxic activity in vitro.The activities of CTL and NK cells,and serum IFN-γlevel were significantly improved,furthermore the tumor growth was obviously suppressed in pcDNA3.1/mIL-21 treated mice.However,the LAK activity did not alter significantly.Taken together,this study suggests that the injection with recombinant plasmid containing mIL-21 is a potential approach for tumor gene therapy.Cellular & Molecular Immunology.2004;1(6):461-466.展开更多
Objective: Interleukin 18 (IL-18) is a strong activator of NK cells and promotes the generation of IL-2, IFN-γ and GM-CSF. In the present study, we constructed adenovirus encoding IL-18 gene (AdIL-18), and observed t...Objective: Interleukin 18 (IL-18) is a strong activator of NK cells and promotes the generation of IL-2, IFN-γ and GM-CSF. In the present study, we constructed adenovirus encoding IL-18 gene (AdIL-18), and observed the biological characteristics of IL-18 gene-modified murine colorectal adenocarcinoma cell (CT26)in vivo andin vitro. Methods: Gene modification was mediated by adenovirus. The proliferation of the cells was determined by MTT and IL-18 was assayed by ELISA. The cytotoxicity of NK and CTL was detected by four-hour51Cr release assay. Results: IL-18 gene modification had no effect on the proliferation and morphology of CT-26 cellsin vitro, but the growth of IL-18-modified CT26 cells was obviously inhibitedin vivo. In addition, although IL-18-modified CT26 cells could form tumor nodulesin vivo as well as LacZ-modified CT26 cells or wild-type CT26 cells, the mean survival time of the mice inoculated with IL-18-modified CT26 cells was significantly prolonged as compared with that of control groups. Thus, the anti-tumor immune responses were induced in the group of mice inoculated with IL-18-modified CT26 cells, which might be related to the activation of NK cells and CTL. However, all the three groups ultimately died of tumor. Conclusion: IL-18-modified CT26 cells could induce the anti-tumor immune responses incompletely, which required other factors for effective anti-tumor responses.展开更多
Oncolysates, debris of tumor cells, have been proven to be effective in active immunotherapy of cancer. In this experiment, the oncolysates from murine melanoma cells B16F10 transfected by recombinant vaccinia viruses...Oncolysates, debris of tumor cells, have been proven to be effective in active immunotherapy of cancer. In this experiment, the oncolysates from murine melanoma cells B16F10 transfected by recombinant vaccinia viruses encoding human IL2(IL2VBO) were used as vaccine. After treatment of tumor bearing mice with pulmonary metastases by intravenous injection of IL2VBO or rVVIL2, higher level IL2 activity was detected in the serum of IL2VBO or rVVIL2 treated mice at 8h. Further experiment results demonstrated that IL2VBO significantly reduced the number of pulmonary metastases and prolonged the survival time of tumorbearing mice when compared with other preparations. Fresh peripheral blood lymphocytes from IL2VBO treated mice showed potent cytotoxicity to B16F10 cells and YAC1 cells. But only cytotoxicity to B16F10 cells is more marked than that in rVVIL2 group, indicating that the IL2VBO could induce specific and non specific antitumor immunity. Because IL2 expression was at the same level in the serum of IL2VBO or rVVIL2 treated mice, the results suggested that the specific antitumor immunity induced by IL2VBO might contribute to the enhanced therapeutic effect of IL2VBO.展开更多
目的：探讨白细胞介素 7裸 DNA质粒用于小鼠肿瘤基因治疗的可行性。方法：构建 IL 7基因真核表达质粒（ pSVK3 IL 7），并将此质粒 DNA直接肌肉注射到 Balb/c小鼠体内，在此之前 7天、同时、之后 7天背部皮下接种 SP 2肿瘤细胞，观察该...目的：探讨白细胞介素 7裸 DNA质粒用于小鼠肿瘤基因治疗的可行性。方法：构建 IL 7基因真核表达质粒（ pSVK3 IL 7），并将此质粒 DNA直接肌肉注射到 Balb/c小鼠体内，在此之前 7天、同时、之后 7天背部皮下接种 SP 2肿瘤细胞，观察该质粒基因的体内抗肿瘤生物学活性；并通过 RT PCR的方法检测注射部位 IL 7mRNA的表达情况。结果：在实验中观察到直接肌肉注射 IL 7基因质粒 DNA的荷瘤鼠，其肿瘤出现的时间明显延迟，肿块也明显小于肿瘤对照组，并且延长了荷瘤鼠的存活时间；基因注射后 7天在注射部位局部有大量的 IL 7mRNA表达。结论：由此看出直接注射裸 DNA IL 7基因质粒有一定的抗肿瘤的效应。展开更多
文摘lnterleukin-21(IL-21)is a recently characterized T cell-derived cytokine with a significant homology to IL-2, IL-4 and IL-15.To determine whether IL-21 has broad immunoregulatory activity and can stimulate durable antitumour responses,we constructed mouse IL-21(mIL-21) recombinant plasmid and evaluated its antitumor efficacy.Mouse IL-21 cDNA was amplified from Con A-activated mouse T cells by RT-PCR.Recombinant pcDNA3.1/mIL-21 was constructed and transfected into Sp2/0 cells.Mouse IL-21 expression was analyzed by Western blotting and its activities were detected by ~3H-TdR incorporation and MTT assay.The recombinant pcDNA3.1/mIL-21 was injected s.c.into tumor lump.Tumor size,weight,the activities of CTLs,NK cells and LAK cells and serum IFN-γ,level were measured for evaluating mIL-21 mediated antitumor responses.The results indicated that mIL-21 was correctly expressed in Sp2/0 cells and it can improve the proliferation of T cells and B cells,and enhance NK cytotoxic activity in vitro.The activities of CTL and NK cells,and serum IFN-γlevel were significantly improved,furthermore the tumor growth was obviously suppressed in pcDNA3.1/mIL-21 treated mice.However,the LAK activity did not alter significantly.Taken together,this study suggests that the injection with recombinant plasmid containing mIL-21 is a potential approach for tumor gene therapy.Cellular & Molecular Immunology.2004;1(6):461-466.
基金National Natural Science Foundation of China (No. 39970689) and Natural Science Foundation of Shanghai (No. 99QB14047).
文摘Objective: Interleukin 18 (IL-18) is a strong activator of NK cells and promotes the generation of IL-2, IFN-γ and GM-CSF. In the present study, we constructed adenovirus encoding IL-18 gene (AdIL-18), and observed the biological characteristics of IL-18 gene-modified murine colorectal adenocarcinoma cell (CT26)in vivo andin vitro. Methods: Gene modification was mediated by adenovirus. The proliferation of the cells was determined by MTT and IL-18 was assayed by ELISA. The cytotoxicity of NK and CTL was detected by four-hour51Cr release assay. Results: IL-18 gene modification had no effect on the proliferation and morphology of CT-26 cellsin vitro, but the growth of IL-18-modified CT26 cells was obviously inhibitedin vivo. In addition, although IL-18-modified CT26 cells could form tumor nodulesin vivo as well as LacZ-modified CT26 cells or wild-type CT26 cells, the mean survival time of the mice inoculated with IL-18-modified CT26 cells was significantly prolonged as compared with that of control groups. Thus, the anti-tumor immune responses were induced in the group of mice inoculated with IL-18-modified CT26 cells, which might be related to the activation of NK cells and CTL. However, all the three groups ultimately died of tumor. Conclusion: IL-18-modified CT26 cells could induce the anti-tumor immune responses incompletely, which required other factors for effective anti-tumor responses.
文摘Oncolysates, debris of tumor cells, have been proven to be effective in active immunotherapy of cancer. In this experiment, the oncolysates from murine melanoma cells B16F10 transfected by recombinant vaccinia viruses encoding human IL2(IL2VBO) were used as vaccine. After treatment of tumor bearing mice with pulmonary metastases by intravenous injection of IL2VBO or rVVIL2, higher level IL2 activity was detected in the serum of IL2VBO or rVVIL2 treated mice at 8h. Further experiment results demonstrated that IL2VBO significantly reduced the number of pulmonary metastases and prolonged the survival time of tumorbearing mice when compared with other preparations. Fresh peripheral blood lymphocytes from IL2VBO treated mice showed potent cytotoxicity to B16F10 cells and YAC1 cells. But only cytotoxicity to B16F10 cells is more marked than that in rVVIL2 group, indicating that the IL2VBO could induce specific and non specific antitumor immunity. Because IL2 expression was at the same level in the serum of IL2VBO or rVVIL2 treated mice, the results suggested that the specific antitumor immunity induced by IL2VBO might contribute to the enhanced therapeutic effect of IL2VBO.
文摘目的：探讨白细胞介素 7裸 DNA质粒用于小鼠肿瘤基因治疗的可行性。方法：构建 IL 7基因真核表达质粒（ pSVK3 IL 7），并将此质粒 DNA直接肌肉注射到 Balb/c小鼠体内，在此之前 7天、同时、之后 7天背部皮下接种 SP 2肿瘤细胞，观察该质粒基因的体内抗肿瘤生物学活性；并通过 RT PCR的方法检测注射部位 IL 7mRNA的表达情况。结果：在实验中观察到直接肌肉注射 IL 7基因质粒 DNA的荷瘤鼠，其肿瘤出现的时间明显延迟，肿块也明显小于肿瘤对照组，并且延长了荷瘤鼠的存活时间；基因注射后 7天在注射部位局部有大量的 IL 7mRNA表达。结论：由此看出直接注射裸 DNA IL 7基因质粒有一定的抗肿瘤的效应。
