Rider V, Jones S, Evans M, Bassiri H, Afsar Z, Abdou N I
School of Biological Sciences, University of Missouri-Kansas City, USA.
J Rheumatol. 2001 Dec;28(12):2644-9.
To examine the in vitro effects of estrogen on CD40 ligand (CD40L) expression in peripheral blood T cells isolated from patients with systemic lupus erythematosus (SLE) and normal controls.
T cells from female patients with SLE and controls were cultured in serum-free medium without and with 2-fluoroestradiol. Some T cells were activated by further culture on anti-CD3 coated plates. Calcineurin was activated in some T cells by culture in ionomycin. Cell surface CD40L was quantitated by FACS analysis. mRNA expression was measured using semiquantitative PCR.
Lupus T cells cultured in medium containing 2-fluoroestradiol showed a significant (p = 0.04) increase in the amount of CD40L on the cell surface, but not in the number of positive cells, compared to the same T cells cultured without estradiol. Estradiol did not significantly change CD40L expression on the surface of T cells from normal women. In addition, the difference in cell surface CD40L between T cells cultured without and with estradiol was significantly greater (p = 0.048) on SLE than on normal T cells. Culture of SLE T cells in medium containing 2-fluoroestradiol followed by T cell receptor (TCR) activation for 2 h using anti-CD3 resulted in a significant (p = 0.04) estrogen dependent increase in CD40L mRNA. The estrogen dependent increases in SLE T cell CD40L mRNA and cell surface protein were blocked by the estrogen receptor antagonist ICI 182,780. SLE and normal T cells pretreated with estradiol and cultured with ionomycin for 2 h to activate calcineurin showed no significant differences in CD40L mRNA.
These results suggest that estradiol, working through the estrogen receptor, stimulates the expression of CD40L in unstimulated and activated SLE T cells. Estradiol effects may be exerted on multiple regulatory steps that control CD40L expression. The estrogen dependent increase in CD40L expression could hyperstimulate SLE T cells and thereby contribute to the pathogenesis of SLE.
研究雌激素对从系统性红斑狼疮（SLE）患者及正常对照者分离出的外周血T细胞中CD40配体（CD40L）表达的体外影响。
将女性SLE患者和对照者的T细胞在无血清培养基中培养，分别添加和不添加2-氟雌二醇。部分T细胞通过在抗CD3包被的平板上进一步培养来激活。部分T细胞通过在离子霉素中培养来激活钙调神经磷酸酶。通过流式细胞术分析定量细胞表面CD40L。使用半定量PCR测量mRNA表达。
与在无雌二醇培养基中培养的相同T细胞相比，在含有2-氟雌二醇的培养基中培养的狼疮T细胞表面CD40L的量显著增加（p = 0.04），但阳性细胞数量未增加。雌二醇对正常女性T细胞表面CD40L表达无显著影响。此外，SLE T细胞在无雌二醇和有雌二醇培养基中培养的细胞表面CD40L差异比正常T细胞显著更大（p = 0.048）。将SLE T细胞在含有2-氟雌二醇的培养基中培养，然后用抗CD3激活T细胞受体（TCR）2小时，导致CD40L mRNA显著（p = 0.04）依赖雌激素增加。雌激素受体拮抗剂ICI 182,780可阻断SLE T细胞中雌激素依赖的CD40L mRNA和细胞表面蛋白增加。用雌二醇预处理并与离子霉素培养2小时以激活钙调神经磷酸酶的SLE和正常T细胞，在CD40L mRNA方面无显著差异。
这些结果表明，雌二醇通过雌激素受体发挥作用，刺激未刺激和激活的SLE T细胞中CD40L的表达。雌二醇的作用可能施加于控制CD40L表达的多个调节步骤。CD40L表达的雌激素依赖性增加可能过度刺激SLE T细胞，从而导致SLE的发病机制。
