Development of monoclonal antibodies against the human 3β-hydroxysteroid dehydrogenase/isomerase isozymes
Development of monoclonal antibodies against the human 3β-hydroxysteroid dehydrogenase/isomerase isozymes
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DOI：
10.1016/j.steroids.2017.08.011
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发表时间：
2017-11-01
期刊：
影响因子：
2.7
通讯作者：
Gomez-Sanchez, Elise P.
中科院分区：
文献类型：
--
作者：
Gomez-Sanchez, Celso E.;Lewis, Mark;Gomez-Sanchez, Elise P.
人β 13-羟基类固醇脱氢酶/异构酶（HSD 3B）催化β 13-羟基δ 5-6类固醇转化为3-酮δ 4-5类固醇，这是合成由肾上腺和性腺分泌的成熟类固醇激素所需的。人类有2种同工酶，传统上位于胎盘和肾上腺外组织的HSD 3B 1和在肾上腺和性腺中表达的HSD 3B 2。隐花色素1和隐花色素2基因缺失的小鼠具有盐敏感性高血压和醛固酮增多症。这些缺失也与Hsd 3b 6酶（人类HSD 3B 1的同源物）在肾小球中的过度表达有关，这被认为是解释醛固酮增多症的原因。一份使用抗人HSD 3B 1抗体的报告表明，它在正常人肾上腺和特发性醛固酮增多症患者的肾小球中表达，而HSD 3B 2在束状肾上腺和肾小球中均表达。我们已经开发了针对人HSD 3B 1和HSD 3B 2同工酶的特异性单克隆抗体，并发现在小球中表达的主要酶是HSD 3B 2。仅在高浓度抗体下使用抗HSD 3B 1抗体也获得肾上腺的微弱染色。本研究未能证实HSD 3B 1在人肾小球中的表达，双重免疫荧光清楚地表明，HSD 3B 2在肾小球和束状体中表达，并且在肾小球中，HSD 3B 2与醛固酮合酶（CYP 11 B2）共表达。
The human 313-hydroxysteroid dehydrogenase/isomerase (HSD3B) enzymes catalyze the conversion of 3j3-hydroxy Delta 5-6 steroids into 3-keto Delta 4-5 steroids, which is required for the synthesis of the mature steroid hormones secreted by the adrenal and gonads. The human has 2 isozymes, the HSD3B1 that is traditionally located in placenta and extra-adrenal tissues and the HSD3B2 that is expressed in the adrenal and gonads. Mice with both cryptochrome 1 and 2 genes deletion were recently found to have salt-sensitive hypertension and hyperaldosteronism. These deletions were also associated with overexpression of the Hsd3b6 enzyme, the homolog of the human HSD3B1, in the zona glomerulosa which was believed to explain the hyperaldosteronism. A report using antibodies against human HSD3B1 suggested that it was expressed in the zona glomerulosa of normal human adrenals and in patients with idiopathic hyperaldosteronism and the HSD3B2 expressed in both the zona fasciculata and glomerulosa. We have developed specific monoclonal antibodies against the human HSD3B1 and HSD3B2 isozymes and found that the main enzyme expressed in the zona glomerulosa was the HSD3B2. Faint staining of the adrenal was also obtained using the anti-HSD3Blantibody only at high concentrations of antibody. This study fails to confirm that HSD3B1 expression in the human zona glomerulosa and double immunofluorescence clearly shows that the HSD3B2 is expressed in the zona glomerulosa and fasciculata and in the zona glomerulosa HSD3B2 is co-expressed with aldosterone synthase (CYP11B2).