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首页> 外文期刊>Journal of Reproductive Immunology >Interleukin-6 and dexamethasone modulate in vitro asymmetric antibody synthesis and UDP-Glc glycoprotein glycosyltransferase activity.
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Interleukin-6 and dexamethasone modulate in vitro asymmetric antibody synthesis and UDP-Glc glycoprotein glycosyltransferase activity.

机译：白介素6和地塞米松调节体外不对称抗体合成和UDP-Glc糖蛋白糖基转移酶活性。

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The increased production of asymmetric IgG protective antibodies is one of the mechanisms proposed to explain a successful semiallogeneic pregnancy. We have previously demonstrated that IL-6 was able to enhance the synthesis of these antibodies by a murine hybridoma, while the glucocorticoid dexamethasone (DEXA) diminished it. In order to investigate the mechanism of asymmetric antibody synthesis, we investigate the role of UDP-Glc glycoprotein glucosyltransferase (GT), an endoplasmic reticulum enzyme involved in the quality control and folding of glycoproteins. Either recombinant murine rmIL-6 (0-10-40-160-320-640 ng/ml) or DEXA (0.15 microM) were added to a mouse hybridoma culture and incubated for 24 and 72 h in the first case, and for 4h in the presence of DEXA. Anti-DNP asymmetric antibodies were determined in the culture supernatants by ELISA. After harvesting, hybridoma cells were sonicated and GT activity was analysed in isolated microsomal fractions by measuring UDP((14)C)-Glc incorporation into urea-denatured thyroglobulin (urea-Tg). In the present paper, we showed that IL-6, mainly at 40 ng/ml and t=24h, was able to upregulate both in vitro GT activity (+74%) and asymmetric molecule synthesis (+227%). Lower increases were obtained employing 10 and 160 ng/ml. On the other hand, DEXA, at 0.15 microM and t=4h, showed a mild inhibition of enzyme activity (-10%) and a diminished proportion of asymmetric IgG (-49%). A direct relationship between GT activity and proportion of the asymmetric antibody synthesised was found in our hybridoma cells employing IL-6 and DEXA in different conditions, as was indicated by a correlation analysis. These results suggest that GT might be involved in the synthesis of asymmetric antibodies.
机译：不对称IgG保护性抗体产量的增加是为解释成功的半同种异体妊娠而提出的机制之一。先前我们已经证明IL-6能够通过鼠类杂交瘤增强这些抗体的合成，而糖皮质激素地塞米松（DEXA）可以将其减少。为了研究不对称抗体合成的机制，我们研究了UDP-Glc糖蛋白葡萄糖基转移酶（GT）的作用，这是一种内质网酶，参与糖蛋白的质量控制和折叠。将重组鼠rmIL-6（0-10-40-160-320-640 ng / ml）或DEXA（0.15 microM）加入小鼠杂交瘤培养物中，在第一种情况下孵育24和72 h，然后孵育4 h在DEXA存在下。通过ELISA在培养上清液中确定抗DNP不对称抗体。收获后，对杂交瘤细胞进行超声处理，并通过测量将UDP（（14）C）-Glc掺入尿素变性的甲状腺球蛋白（urea-Tg）中来分析分离的微粒体组分中的GT活性。在本文中，我们显示IL-6主要在40 ng / ml和t = 24h时，能够上调体外GT活性（+ 74％）和不对称分子合成（+ 227％）。使用10和160 ng / ml获得较低的增加。另一方面，DEXA在0.15 microM和t = 4h时显示出对酶活性的轻度抑制（-10％）和不对称IgG的比例减少（-49％）。相关性分析表明，在不同条件下使用IL-6和DEXA的杂交瘤细胞中，GT活性与合成的不对称抗体比例之间存在直接关系。这些结果表明GT可能参与不对称抗体的合成。

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