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P450 Side-Chain Cleavage Enzyme (P450-SCC) Is an Ovarian Autoantigen in a Mouse Model for Autoimmune Oophoritis

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Abstract

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.

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Data Availability

The data and materials are available. The software used in this study is available online for the public.

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Acknowledgements

We are thankful to Dr. Lawrence Nelson for valuable discussion of this study and helpful improvement of the manuscript, and to Ms. Nancy Terry at NIH Library for providing an editing service on this manuscript. We appreciate Dr. David Gerhold and Dr. Anton Simeonov for their critical readings of the manuscript.

Funding

This work was supported by the Intramural Research Programs at NICHD and NCATS, National Institutes of Health, USA.

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Authors and Affiliations

  1. Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, 9800 Medical Center Drive, Rockville, MD, 20850, USA

    Zhi-Bin Tong

  2. Reproductive Biology and Medicine Branch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA

    Zhi-Bin Tong, Noriyuki Otsuka, Wei Tu, Qingxiang Wei & Alan H. DeCherney

  3. Department of Pathology, Graduate School of Medicine/Faculty of Medicine, Hokkaido University, Sapporo, Japan

    Noriyuki Otsuka

  4. Division of Viral Products, CBER, FDA, Silver Spring, MD, 20993, USA

    Wei Tu

  5. Global Biopharma Development, Incyte, Wilmington, DE, 19803, USA

    Qingxiang Wei

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  1. Zhi-Bin Tong
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  2. Noriyuki Otsuka
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Contributions

ZBT and AHD conceived and designed the study. ZBT, NO, WT, and QW carried out the experiments. ZBT drafted the manuscript. All authors participated in the process of the manuscript’s reviews and edits. All authors approved the manuscript for a submission and publication.

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Correspondence to Zhi-Bin Tong.

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This study was performed under the animal study protocols approved by the animal care and study commission at NICHD.

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The authors declare no competing interests.

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Tong, ZB., Otsuka, N., Tu, W. et al. P450 Side-Chain Cleavage Enzyme (P450-SCC) Is an Ovarian Autoantigen in a Mouse Model for Autoimmune Oophoritis. Reprod. Sci. 29, 2391–2400 (2022). https://doi.org/10.1007/s43032-022-00970-7

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