中华皮肤科杂志 ›› 2002, Vol. 35 ›› Issue (6): 425-428.

• 论著 • 上一篇    下一篇

干扰素γ诱导人黑素细胞CD40的表达

鲁严1, 朱文元1, 谭城1, 於葛华2, 顾劲雄2   

  1. 1. 南京医科大学第一附属医院皮肤科 210029;
    2. 苏州大学医学院免疫学教研室
  • 收稿日期:2001-11-12 出版日期:2002-12-15 发布日期:2002-12-15
  • 基金资助:
    国家自然科学基金资助(39870701)

CD40 Expression on Melanocytes Induced by IFN-γ and Its Significance

LU Yan1, ZHU Wenyuan1, TAN Cheng1, YU Gehua2, Gu Jinxiong2   

  1. Department of Dermatology, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China
  • Received:2001-11-12 Online:2002-12-15 Published:2002-12-15

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摘要： 目的 探讨γ-干扰素(IFN-γ)体外诱导人黑素细胞表面CD40分子的表达及其意义.方法 常规分离培养人黑素细胞,流式细胞仪测定IFN-γ处理前后细胞表面CD40等免疫分子表达;混合淋巴细胞反应评价黑素细胞对同种异体T淋巴细胞的刺激能力;ELISA检测培养上清液IL-8、IL-10、IL-12的浓度.结果 体外培养的人黑素细胞表面表达少量CD40分子;不同浓度的IFN-γ处理黑素细胞24h、48h、72h后,能显着促进其CD40的上调(P<0.01),且24h处理组的黑素细胞CD40的表达量和IFN-γ的浓度呈直线相关.经IFN-γ诱导后的黑素细胞形态有所变化,刺激同种异体淋巴细胞的能力也显着增加,300IU/mLIFN-γ处理的黑素细胞72h刺激指数(SI)可达到峰值.黑素细胞经IFN-γ作用后,培养上清液中IL-12水平明显增加(P<0.05),而IL-8、IL-10的浓度无变化(P>0.05).经IFN-γ预处理的黑素细胞经SCD40L配基化后,能显着上调CD80、细胞间粘附分子1(P<0.01),且这种作用能被特异性的CD40L的单克隆抗体所阻断.结论 IFN-γ体外能够诱导黑素细胞功能性地表达CD40分子及增加对淋巴细胞的刺激能力,这对于认识黑素细胞在细胞免疫应答中的作用具有重要的意义,CD40分子上调后,黑素细胞可能不经过CD4+细胞而直接刺激活化CD8+的杀伤性T细胞(CTL).

关键词: 干扰素γ,重组, 黑素细胞, 抗原,CD40

Abstract: Objective To study CD40 expression on melanocytes induced by IFN-γ and its significance. Methods CD40 expression was detected by flow cytometry. The capacity of melanocytes to stimulate T lymphocytes was evaluated by mixed ly mphocyte reaction and the supernatant cytokine levels were determined by ELISA. Results Human melanocytes(MC) cultured in vitro expressed low but detectable CD40 surf ace protein. The surface expression of CD40 was markedly up-regulated by stimul ation with interferon(IFN)-γ with different concentrations for 24 hours,48 hours and 72 hours, respectively. The expression of CD40 was correlated with IFN-γ levels after 24 hour incubation. MC underwent a morphologic change with an increased capacity to stimulate allogenic lymphocytes to proliferate after IFN-γ stimulation. Optimal enhancement of stimulating index(SI) was observed at an IFN-γ concentration of 300 IU/ml after 72 hour treatment. Meanwhile concentrations of interleukin-12 but not interleukin-8 or 10 were obviously increased in the supernatants of cultured MC. Furthermore, ligation of CD40 via soluble CD40 ligan d(SCD40L) could enhance CD80 and ICAM-1 expression, which could be blocked by specific monoclonal antibody to CD40L. Conclusions Since CD40-CD40L is a pair of important and special costimulating signal, it is of great value to elucidate t he fact that CD40 is functionally expressed on MC, thus for a better understandi ng of MC's role in cellular immune responses. MC mightactivate cytotoxic T lymphocyte directly and not via CD4 positive lymphocyte.

Key words: Interferon-gamma, recombinant, Melanocytes, Antigens, CD40