In vitro modulation of protective antibody responses by estrogen, progesterone and interleukin-6
In vitro modulation of protective antibody responses by estrogen, progesterone and interleukin-6
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DOI：
10.1034/j.1600-0897.2002.01141.x
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发表时间：
2002-11-01
影响因子：
3.6
通讯作者：
Idehu, RAM
中科院分区：
文献类型：
--
作者：
Canellada, A;Blois, S;Idehu, RAM
问题：我们之前已经证明，在小鼠杂交瘤中添加胎盘白介素-6 （IL-6）会增加不对称抗体的合成。本研究分析了黄体酮（Pg）和雌激素（E-2）是否通过调节IL-6的产生影响杂交瘤细胞的不对称抗体合成。研究方法：用E-2、Pg或重组IL-6培养杂交瘤112d5b细胞。采用3 h -胸腺嘧啶掺入法检测细胞增殖，采用Con A固定和酶联免疫吸附试验（ELISA）检测培养上清中的不对称抗体。Western blot检测全细胞提取物中E-2和pg受体（ER和PR）的表达。ELISA法检测培养上清液中IL-6的含量。结果：112135同时表达PR和ER，并有差异调控。48小时时，Pg和E-2轻微抑制细胞增殖，而IL-6没有。以及IL-6、10(-10)m Pg而非E-2诱导不对称抗体的产生。有趣的是，Pg在10(-6)m时减少了杂交瘤细胞的不对称抗体合成。最后，以Pg为主，E-2也增加。结论：在同时表达ER和PR的细胞中，我们可以证明类固醇通过调节不对称抗体的合成参与了体液免疫反应。IL-6被证明只是部分参与。本文还讨论了其他可能的机制，包括Pg对阻断抗体反应的影响及其对成功妊娠的贡献。
PROBLEM: We have previously demonstrated that the addition of placental interleukin-6 (IL-6) to murine hybridomas increased asymmetric antibody synthesis. Here we analyze whether progesterone (Pg) and estrogen (E-2) affect asymmetric antibody synthesis by modulating IL-6 production in hybridoma cells.METHOD OF STUDY: Hybridoma 112D5 B cells were cultured with E-2, Pg or recombinant IL-6. Cell proliferation was assessed by 3 H-thymidine incorporation, and asymmetric antibodies were measured in culture supernatants by Con A fixation and enzyme-linked immunusorbant assay (ELISA). E-2 and Pg-receptors (ER and PR) were evaluated in whole cell extracts by Western blot. IL-6 was measured in culture supernatants by ELISA.RESULTS: 112135 expressed both PR and ER, which were differentially regulated. At 48 hr, Pg and E-2 slightly decreased cell proliferation whereas IL-6 did not. As well as IL-6, 10(-10) m Pg but not E-2 induced asymmetric antibody production. Interestingly, Pg at 10(-6) m decreased asymmetric antibody synthesis by hybridoma cells. Finally, mainly Pg but also E-2 increased. IL-6 synthesis, although IL-6 levels did not correlate with asymmetric antibodies synthesized in the presence of E-2 or Pg.CONCLUSIONS: In cells expressing both ER and PR, we could demonstrate that steroids participate in humoral immune responses by modulating asymmetric antibody synthesis. IL-6 proved to be only partially involved. Other possible mechanisms involved in the effect of Pg on blocking antibody responses and their contribution to a successful pregnancy are discussed in the paper.