中华皮肤科杂志 ›› 2001, Vol. 34 ›› Issue (4): 277-279.

• 论著 • 上一篇    下一篇

小鼠角质形成细胞MHC-Ⅰ类限制性内源性抗原递呈系统的构建

高兴华1, 小阪博2, 陈洪铎1, 吉川邦彦2   

  1. 1. 中国医科大学第一临床学院皮肤科 沈阳 110001;
    2. 日本大阪大学医学部皮肤科
  • 收稿日期:2000-11-18 出版日期:2001-08-15 发布日期:2001-08-15
  • 基金资助:
    国家自然科学基金(39970682)

Construction of a MHC ClassⅠ Restricted Endogenous Antigen Presenting System in Mouse Kera tinocytes

GAO Xinghua1, Hiroshi Kosaka2, CHEN Hong-Duo1   

  1. Department of Dermatology, No.1 Hospital of China Medical University, Shenyang 110001, China
  • Received:2000-11-18 Online:2001-08-15 Published:2001-08-15

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摘要： 目的 建立一转染小鼠角质形成细胞系,使其表达MHC-Ⅰ类分子限制的卵白蛋白(ovalbumin,OVA)抗原。方法 将含有融合OVA基因(在角蛋白5的启动子下)和小鼠H-2Kb的重组质粒转染于小鼠PAM212细胞系(H-2Kd)。采用RT-PCR和流式细胞分析技术检测相应基因的转录及翻译水平的表达。结果 本实验建立了2个可表达小鼠H-2Kb分子(命名为PAM-Kb)的转染细胞系,4个可表达OVAmRNA(命名为PAM-OVA)的转染细胞系和2个可同时表达小鼠H-2Kb分子和OVAmRNA的转染细胞系(命名为PAM-OK)。结论 所建立的MHC-Ⅰ限制的内源性抗原系统为进一步理解多肽/MHC-Ⅰ复合体与具有相应特异T细胞受体的CD8+T细胞之间的相互作用奠定了基础。

关键词: 角蛋白细胞, 卵白蛋白, 主要组织相容性复合物

Abstract: Objective To set up a mouse keratinocyte line that expresses a H-2Kb restricted endogenous ovalbumin (OVA) antigen. Methods A mouse keratinocyte line (PAM212, H-2Kd) was transfected with a recombinant plasmid containing a desired partial sequence of ovalbumin under K5 promotor, and/or with other recombinant plasmid containing a H-2Kb gene. Cloned transfected PAM212 cells were examined by RT-PCR for OVA mRNA expression and flow cytometry for H-2Kb molecule expression. Results PAM212 transfectant lines that expressed OVA mRNA (named PAM-OVA, 4 lines), H-2Kb molecule (PAM-Kb, 2 lines) and both OVA mRNA and H-2Kb molecule (PAM-OK, 2 lines) were set up. Conclusion PAM-OK cell line will serve as a useful tool for interpreting interactions between peptide/MHC-Ⅰ complex and TCR specific CD8+ T cells.

Key words: Keratinocytes, Ovalbumin, Major histocompatibility complex