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《Chinese Journal of Cancer Biotherapy》 2018-03
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Effects of IL-7 and IL-21 modified NK-92MI cells on themselves and T cells from normal human peripheral blood

ZHANG Ping;LI Yafen;AN Gangli;YANG Lin;The Cyrus Tang Hematology Center, Medical department, Soochow University;Persongen Bio Therapeutics (Suzhou) Co., Ltd.;  
Objective:To investigate whether the proliferation and cytotoxicity of NK-92MI cells can be improved by IL-7 and IL-21 genes modification, and determine the effects of this genetically modified NK-92MI cells on T cells from normal human peripheral blood. Methods:IL-7 and IL-21 gene fragments were constructed into electroporation vector by genetic engineering method, and NK-92MI/IL-21 and NK-92MI/IL-721 cells were constructed by electroporation transfection. The in vitro proliferation and cytotoxicity of NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-721 cells were measured by cell count and flow cytometry assays. Then, normal human PBMCs were co-cultured with NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-721 cells in vitro respectively, and the phenotype change of T cells was measured by flow cytometry. In addition, the cytotoxicity between the activated T cells and three NK-92MI cell lines(NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-721 cells) as well as the cytotoxicity of the three NK-92MI cells on tumor cells after co-incubation with activated T cells were detected. Results: NK-92MI/IL-21 cell line(highly expressing IL-21) and NK-92MI/IL-721 cell line(highly expressing both IL-7 and IL-21) were successfully constructed. The toxicity of NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-721 cells on Jurkat and K562 cells showed no difference, while the proliferation of NK-92MI/IL-21 and NK-92MI/IL-721 cells was increased compared with NK-92MI cells. Furthermore, NK-92MI/IL-21 and NK-92MI/IL-721 cells promoted the activation of T cells to a certain degree, and the activated T cells showed merely no cytotoxicity on NK-92MI, NK-92MI/IL-21 and NK-92MI/IL-721 cells; Meanwhile, the activated T cells did not affect the cytotoxicity of the three NK cells(NK-92MI, NK-92MI/IL-21, and NK-92MI/IL-721 cells) on K562 cells under their co-existence. Conclusion: The in vitro proliferation of NK-92MI/IL-21 and NK-92MI/IL-721 cells were enhanced after gene modification, which could also stimulate and promote the activation of T cells from peripheral blood. The cytotoxicity assay showed that the activated T cells had no cytotoxicity on NK-92MI, NK-92MI/IL-21, and NK-92MI/IL-721 cells. Meanwhile, the presence of the activated T cells did not affect the cytotoxicity of NK-92MI cells.
【Fund】: 国家重点研发计划(No.2016YFC1303403);; 国家自然科学基金资助项目(No.31471283);; 江苏高校血液学协同创新中心(No.XYXT2015304);; 江苏高校优势学科建设工程资助项目~~
【CateGory Index】: R730.51
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