中华皮肤科杂志 ›› 2002, Vol. 35 ›› Issue (6): 449-451.

• 论著 • 上一篇    下一篇

UVB与钙离子对体外培养人角质形成细胞表达天疱疮抗原的影响

渠涛, 王宝玺, 邵燕玲, 曾建英   

  1. 中国医学科学院、中国协和医科大学北京协和医院皮肤科 北京 100730
  • 收稿日期:2001-11-29 出版日期:2002-12-15 发布日期:2002-12-15

Effects of UVB Irradiation and Calcium on the Expression of Pemphigus Antigens by Human Keratinocytes

QU Tao, WANG Baoxi, SHAO Yanling, ZENG Jianying   

  1. Department of Dermatology, Pe king Union Medical College Hospital, Peking Union Medical College, Chinese Acade my of Medical Sciences, Beijing 100730, China
  • Received:2001-11-29 Online:2002-12-15 Published:2002-12-15

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摘要： 目的 研究中波紫外线(UVB)照射以及不同钙离子浓度对角质形成细胞表达天疱疮抗原的影响.方法 通过改变培养基中的钙离子浓度以及采用不同剂量UVB照射体外培养的人角质形成细胞,在不同时间段以寻常型天疱疮(PV)或落叶型天疱疮(PF)血清作为一抗进行免疫荧光检测,观察寻常型天疱疮抗原(PVA)和落叶型天疱疮抗原(PFA)的表达情况;提取细胞或皮肤表皮组织蛋白质,用PV和PF血清进行免疫印迹检测.结果 无论是否增加钙离子浓度,体外培养人角质形成细胞间隙均可以检测到PV血清特异性着色.只有增加培养基中钙离子浓度后,形成的复层化角质形成细胞间隙才可以检测到PF血清特异性荧光着色.不同剂量UVB照射后的角质形成细胞均不产生PF血清的特异性着色.PF血清与160000条带反应,PV血清可与130000和160000两条带反应.结论 体外培养的单层或复层角质形成细胞均可以表达PVA,提高培养基中钙离子浓度可以诱导培养人角质形成细胞复层化并表达PFA,而UVB照射不能促使人角质形成细胞在体外培养条件下表达PFA.

关键词: 角蛋白细胞, 天疱疮, 钙, 紫外线

Abstract: Objective To investigate the effects of UVB irradiation and calcium at different concentrations on the expression of pemphigus antigens by culture d human keratinocytes. Methods Human keratinocyte cultures were treated witheit her 2 mmol/L calcium added to the serum free media, or exposure to UVB irradiation. Immunofluorescence staining was performed with sera from patients with pemp higus vulgaris (PV) or pemphigus foliaceus (PF) as the first antigen. Extracts from both epidermis and keratinocytes were run on SDS-PAGE according to Laemmli's method and transferred to nitrocellulose membrane for immunoblot with PV and PF sera. Results Specific staining with PV sera was always detectable on kerati nocyte culture by immunofluorescence assay with or without high concentrations o f calcium while PF antigen was detected on stratified cells only. However, exposure to UVB irradiation could not evoke keratinocyte culture express PF antigen. The reactivities were found at 160 kd band with PF sera while at both 160 kd and 130 kd bands with PV sera. Conclusions Monolayer or stratified keratinocytes can produce PV antigen, by increasing the concentration of calcium in the culture media, the human cultured kera tinocytes can be induced to stratify and express PFA. Human keratinocytes can not express PF antigen after exposure to UVB in intro.

Key words: Keratinocytes, Pemphigus, Calcium, Ultraviolet rays