中华皮肤科杂志 ›› 2009, Vol. 42 ›› Issue (1): 5-7.

• 论著 • 上一篇    下一篇

大疱性类天疱疮疱液中IgG对角质形成细胞分泌趋化因子的影响

袁艳霞 林麟 周武庆 康定华 冯素英   

  1. 常州第一人民医院皮肤科 南京医科院皮研所 南京医科院皮研所 江苏省常州市第一人民医院皮肤科 南京医科院皮研所
  • 收稿日期:2008-03-24 修回日期:2008-04-21 出版日期:2009-01-15 发布日期:2009-01-15
  • 通讯作者: 袁艳霞 E-mail:doctoryanxia@yahoo.com.cn

Influence of IgG in bullous pemphigoid blister fluid on the secretion of chemokines by human kera- tinocytes

  • Received:2008-03-24 Revised:2008-04-21 Online:2009-01-15 Published:2009-01-15

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摘要：

目的 探讨大疱性类天疱疮IgG（BP IgG）对角质形成细胞分泌趋化因子的影响。方法 应用辛酸－硫酸铵法从BP患者疱液中纯化IgG，并用BP180 ELISA试剂盒鉴定疱液纯化前后的免疫活性。将纯化的BP IgG与角质形成细胞共同孵育，以正常人血清纯化后IgG作对照。设置0.5、1、2、4、8 g/L IgG浓度组，收集培养24 h后上清液；再以含相同浓度（4 g/L）BP IgG和正常人IgG的培养液进行培养，收集培养3、6、12、24 h后上清液。用ELISA法检测培养上清液中嗜酸粒细胞趋化因子、单核细胞趋化蛋白-1和IL-8的表达情况。结果 用辛酸-硫酸铵法从BP患者疱液中纯化IgG，不影响提纯前后的免疫活性。纯化的BP IgG以时间依赖方式、浓度依赖方式刺激角质形成细胞分泌IL-8，与正常人血清纯化后IgG刺激组相比，差异具有统计学意义（P均 ＜ 0.01）。介质组、正常人IgG刺激组和BP IgG刺激组的培养上清液，均未检测到嗜酸粒细胞趋化因子和单核细胞趋化蛋白-1。结论 BP IgG能刺激角质形成细胞分泌IL-8，这一过程可能参与BP的发病。

关键词: 类天疱疮，大疱性;炎症趋化因子类;角蛋白细胞

Abstract:

Objective To investigate the effect of immunoglobulin G （IgG） in bullous pemphigoid （BP） blister fluid on the secretion of chemokines by human keratinocytes. Methods IgG was obtained from the blister fluid of patients with bullous pemphigoid and sera of normal human controls, then purified by sequential precipitation with caprylic acid and ammonium sulfate. The immunological activity of blister fluid was tested before and after the purification by BP180 ELISA kit. Keratinocytes were isolated from the foreskin tissue of yong adults, and subjected to primary culture. After 3 passages, the primary keratinocytes were harvested and subcultured in the presence of purified IgG of 0.5, 1, 2, 4 and 8 g/L, respectively, for 24 hours, or IgG of 4 g/L for 3, 6, 12, 24 hours, respectively, followed by the detection of levels of eotaxin, monocyte chemoattractant protein （MCP）-1 and interleukin （IL）-8 in the supernate of keratinocytes by ELISA. Results The valence of IgG remained unchanged after the purification with caprylic acid and ammonium sulfate. Compared with IgG from the sera of normal controls, that from bullous pemphigoid blister fluid significantly enhanced the secretion of IL-8 by keratinocytes in a time- and dose-dependent manner （both P < 0.01）. Neither eotaxin nor MCP-1 was detected in the supernate of control IgG-treated, BP IgG-treated or untreated keratinocytes. Conclusions The IgG in BP blister fluid has been proved to stimulate the secretion of IL-8 by cultured human keratinocytes, which may be involved in the pathogenesis of BP.

Key words: Pemphigoid, bullous;Chemokines;Keratinocyte