Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, 9800 Medical Center Drive, Rockville, MD, 20850, USA.
Reproductive Biology and Medicine Branch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA.
Reprod Sci. 2022 Aug;29(8):2391-2400. doi: 10.1007/s43032-022-00970-7. Epub 2022 May 18.
Steroid-producing cells contain key cytochrome P450 enzymes, such as side-chain cleavage (P450-SCC) and 17α-hydroxylase (17α-OH). They are required for steroid hormone synthesis and considered antigens associated with Addison's disease and autoimmune primary ovarian insufficiency (POI). We studied an animal model for human autoimmune POI in mice with autoimmune oophoritis induced by neonatal thymectomy performed at day 3 (TX3). We previously identified an oocyte-specific protein as a major antigen inciting autoimmune oophoritis in mice. In this study, we characterized ovarian steroid-producing cell antigens. Using indirect immunofluorescence staining, we tested immune reactions in mouse ovarian and adrenal tissue sections with sera from TX3 female mice. More than half of the TX3 mice (8 of 15) produced antibodies reacting with both ovarian and adrenal steroid-producing cells, including some that reacted to oocytes as well. We produced recombinant proteins for the three key steroidogenic enzymes 17α-OH, P450-SSC, and 3β-hydroxysteroid dehydrogenase (3β-HSD) and tested their immune reactions with individual mouse sera. By immunoblotting, all mouse sera that reacted with the steroid-producing cells (n = 8) were shown to react with the P450-SCC, but not with the 17α-OH or 3β-HSD recombinant proteins. The sham-operated mouse sera and TX3 mouse sera negative for steroid-producing cells did not react with the P450-SCC recombinant protein. Our findings indicate that the P450-SCC is a specific and unique major antigen within the ovarian steroid-producing cells. Given their similarity of predicted antigenicity, we assume that P450-SCC acts in human autoimmune POI as it does in mouse autoimmune oophoritis.
产类固醇细胞含有关键的细胞色素 P450 酶，如侧链裂解酶（P450-SCC）和 17α-羟化酶（17α-OH）。它们是类固醇激素合成所必需的，被认为与艾迪生病和自身免疫性原发性卵巢功能不全（POI）相关的抗原。我们在通过在第 3 天（TX3）进行新生胸腺切除术诱导的自身免疫性卵巢炎的小鼠中研究了人类自身免疫性 POI 的动物模型。我们之前鉴定了一种卵母细胞特异性蛋白，它是激发小鼠自身免疫性卵巢炎的主要抗原。在这项研究中，我们对卵巢产类固醇细胞的抗原进行了特征分析。使用间接免疫荧光染色，我们用 TX3 雌性小鼠的血清对小鼠卵巢和肾上腺组织切片进行了免疫反应测试。超过一半的 TX3 小鼠（15 只中的 8 只）产生了与卵巢和肾上腺产类固醇细胞反应的抗体，其中一些也与卵母细胞反应。我们为三种关键的类固醇生成酶 17α-OH、P450-SCC 和 3β-羟类固醇脱氢酶（3β-HSD）生成了重组蛋白，并测试了它们与单个小鼠血清的免疫反应。通过免疫印迹，与产类固醇细胞反应的所有 8 只小鼠血清（n=8）均与 P450-SCC 反应，但与 17α-OH 或 3β-HSD 重组蛋白不反应。未与产类固醇细胞反应的假手术操作小鼠血清和 TX3 小鼠血清均不与 P450-SCC 重组蛋白反应。我们的研究结果表明，P450-SCC 是卵巢产类固醇细胞内的一种特异性和独特的主要抗原。鉴于它们预测的抗原性相似，我们假设 P450-SCC 在人类自身免疫性 POI 中的作用与在小鼠自身免疫性卵巢炎中相同。
