中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (5): 297-299.

• 论著 • 上一篇    下一篇

金黄色葡萄球菌肠毒素A基因在C57BL/6小鼠淋巴细胞中的表达

汪宇1, 陆洪光1, 程波1, 麦跃2, 余德厚1   

  1. 1. 贵阳医学院附属医院皮肤科, 550004;
    2. 第三军医大学防原教研室
  • 收稿日期:2004-05-24 出版日期:2005-05-15 发布日期:2005-05-15
  • 通讯作者: 陆洪光,E-mail:hongguanglu@yahoo.com E-mail:hongguanglu@yahoo.com
  • 基金资助:
    国家自然科学基金资助项目(39960071)

Expression of SEA Gene in Lymphocytes of C57BL/6 Mice Mediated by Replicative-deficient Recombinant Adenovirus Vector

WANG Yu1, LU Hong-guang1, CHENG Bo1, MAI Yue2, YU De-hou1   

  1. Department of Dermatology, Affiliated Hospital of Guiyang Medical College, Guiyang 550004, China
  • Received:2004-05-24 Online:2005-05-15 Published:2005-05-15

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摘要： 目的 通过复制缺陷型腺病毒载体的介导,将金黄色葡萄球菌肠毒素A(SEA)基因转染入C57BL/6小鼠淋巴细胞中,观察基因表达产物对B16细胞的作用。方法 通过MTT法直接测定重组腺病毒颗粒感染的淋巴细胞的增殖情况,采用双抗体夹心ABC-ELISA法,测定淋巴细胞培养上清液中白介素2的水平。MTT法测定活细胞数目,以了解活化的C57BL/6小鼠淋巴细胞对B16细胞的杀伤效应。结果 加入不同滴度的重组腺病毒颗粒后,C57BL/6小鼠淋巴细胞明显增殖;同时淋巴细胞培养上清液中白介素2水平也明显增加。重组腺病毒颗粒转染的C57BL/6小鼠淋巴细胞可明显杀伤B16细胞,效/靶比越高,杀伤效应越明显。结论 SEA基因可以在C57BL/6小鼠淋巴细胞中表达,并且表达产物能够活化C57BL/6小鼠淋巴细胞,进而杀伤B16细胞。

关键词: 基因,细菌, 遗传载体, 基因表达, 致细胞病变,病毒

Abstract: Objective To investigate the effects of staphylococcal enterotoxin A SEA gene on target cells mediated by replicative-deficient recombinant adenovirus vector. Methods Lymphocytes of C57BL/6 mice were infected with various titers of recombinant adenoviruses. Supernatants were collected after 12 h, 24 h, 48 h, 72 h, 96 h, 120 h and 144 h of incubation and analyzed for proliferation of lymphocytes by MTT assay. IL-2 level in the culture supernatants was measured with ELISA. The killing effect of lymphocytes was also observed by MTT assay. Results Proliferation response and elevated levels of IL-2 were observed in experimental group. The killing effect on B16 cells was stronger in experimental group, which seemed to be dose-dependent with the increase of ratio of lymphocytes/target cells. Conclusions SEA gene can be expressed in lymphocytes of C57BL/6 mice mediated by replicative-deficient recombinant adenovirus vector. The expressing products can activate lymphocytes of C57BL/6 mice, which kill B16 cells in vitro.

Key words: Genes, bacterial, Genetic vectors, Gene expression, Cytopathogenic effect, virus