中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (11): 788-791.

• 论著 • 上一篇    下一篇

双链DNA抗原冲激导致髓源性树突细胞免疫表型变化

夏育民1,方春红2,江珊3,程鸿3   

  1. 1. 武汉大学人民医院皮肤科江珊收转13659892065
    2. 武汉大学人民医院皮肤科
    3.
  • 收稿日期:2010-03-15 修回日期:2010-03-31 出版日期:2010-11-15 发布日期:2010-11-10
  • 通讯作者: 夏育民 E-mail:xiayumin1202@163.com
  • 基金资助:

    国家自然科学基金

Exogenous double-stranded DNA induces immunophenotypic changes of bone marrow-derived dendritic cells

  • Received:2010-03-15 Revised:2010-03-31 Online:2010-11-15 Published:2010-11-10
  • Contact: Yu-min XIA E-mail:xiayumin1202@163.com

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摘要：

目的 探索外源性双链DNA物质对小鼠髓源性树突细胞（DC）免疫表型的影响。方法 采用免疫磁珠法分离C57小鼠骨髓lin-CD117+ 干细胞，用多种细胞因子诱导后其增殖并发育成不同成熟阶段的DC。提取马疫锥虫动基体DNA（kDNA），对上述DC进行冲激。采用流式细胞法和激光共聚焦显微镜检测DC免疫表型和形态学变化。结果 冲激前，未成熟、半成熟和成熟DC的MHCⅡ阳性率依次为11.42% ± 2.56%、27.08% ± 5.29%与44.63% ± 10.37%，CD80阳性率为8.54% ± 2.01%、31.35% ± 6.40%与52.96% ± 10.34%，CD86阳性率为10.22% ± 3.47%、32.15% ± 6.83%与64.72% ± 9.68%。冲激后，这三组DC的MHCⅡ阳性率分别上升15.63%、9.66%、4.12%，与冲激前比较，t值分别为6.21、4.35与2.82，P值均 < 0.05；CD80阳性率上升9.63%、7.09%与4.09%，CD86阳性率上升13.16%、9.75%与3.10%，升高幅度皆为未成熟DC > 半成熟DC > 成熟DC。结论 双链DNA抗原可促进髓源性DC表达成熟免疫表型，且成熟程度越低的DC受影响越显著。

关键词: 红斑狼疮, 抗体，抗双链DNA, 树突状细胞, 免疫表型

Abstract:

Objective To study the effects of exogenous double-stranded DNA antigen on the immunophenotypic changes of dendritic cells （DCs） derived from stem cells in mouse bone marrow. Methods Lin-CD117（c-kit）+ hemopoietic stem cells were obtained from the bone marrow of C57 mice by magnetic affinity cell sorting. Some cytokines, including granulocyte-macrophage colony-stimulating factor, interleukin-4, tumor necrosis factor-α and so on, were used to enhance the proliferation or differentiation of stem cells to obtain mature, semimature and immature DCs. The double stranded DNA of kinetoplast （kDNA） was isolated from Trypanosoma equiperdum, and added to the culture media to pulse DCs. The immunophenotypic and morphologic features of DCs were analyzed by using flow cytometry and laser confocal microscopy respectively. Results The expression rates of CD117 and CD11c in DCs showed no significant changes after kDNA pulse compared with those before the pulse. In unpulsed immature, semi-mature and mature DCs, the expression rate was 11.42% ± 2.56%, 27.08% ± 5.29% and 44.63% ± 10.37% for MHCⅡ, 8.54% ± 2.01%, 31.35% ± 6.40% and 52.96% ± 10.34% for CD80, 10.22% ± 3.47%, 32.15% ± 6.83% and 64.72% ± 9.68% for CD86, respectively. After pulse with the kDNA antigen, the expression rate increased by 15.63%, 9.66% and 4.12% （t = 6.21, 4.35, 2.82, P < 0.05） for MHCⅡ, by 9.63%, 7.09% and 4.09% for CD80, by 13.16%, 9.75% and 3.10% for CD86, respectively in immature, semi-mature and mature DCs, respectively. The increase of expression rate of these membrane antigens in decreasing order was observed in immature DCs, semi-mature DCs and mature DCs. Conclusions The exogenous DNA antigen could enhance the maturation of bone marrow-derived DCs, likely by upregulating the expression of certain immunophenotypic membrane proteins, and the lower the maturity degree, the more liable the DCs to be affected by the antigen.

Key words: Lupus erythematosus, Antibodies, anti-double stranded DNA, Dendritic cells, Immunophenotype