Evaluation of the epididymis following intravasal-epididymal injection of SARS-CoV-2 antigens focusing on the clear cell response

• Grant number: 23/11482-8
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: April 01, 2024
• End date: September 30, 2024
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: Estela Sasso Cerri
• Grantee: André Acácio Souza da Silva
• Supervisor: Maria Agustina Battistone
• Host Institution: Faculdade de Odontologia (FOAr). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Institution abroad: Massachusetts General Hospital, United States
• Associated to the scholarship: 22/10560-2 - Immunological response of epididymis of K18-hACE2 transgenic mice infected with SARS-CoV-2, with emphasis on the clear cell, BP.MS

Abstract

The global pandemic COVID-19, caused by SARS-CoV-2, lead to millions of deaths worldwide, and studies have shown that men were more susceptible to infection. The epididymis is an essential organ for male fertility since the specialized luminal environment provides sperm motility and fertilizing capacity. Clear cells, one of the most abundant cells in the epididymis, regulate the luminal fluid pH through V-ATPase recruitment to the apical membrane. However, recent studies performed by Battistone and collaborators have demonstrated that these cells express pro-inflammatory cytokines and chemokines in response to LPS injection and that these cells may have a close correlation with macrophages and dendritic cells present in the epididymis, playing an immunoregulatory role in the epididymis. In our study with transgenic K18-hACE2 mice infected with SARS-CoV-2, we have shown increased expression of TNF-± in epididymal clear cells and an increase in intraepithelial macrophages in all portions of the epididymis. Thus, the aim of this study is to evaluate, with the collaboration of Dr. M. Agustina Battistone, the immune response of epididymis with emphasis on clear cells following intravasal-epididymal injection of SARS-CoV-2 purified lysate. Transgenic animals expressing clear cell markers (ATP6V1B1-EGFP) and mononuclear phagocyte markers (CXCR1-EGFP or CD11c-YFP) will be used. Using the intravasal-epididymal injection technique performed in the Dr. Battistone's lab, SARS-CoV-2 purified lysates will be injected in the epididymal lumen, and the RNA-seq of clear cells and mononuclear phagocytes will be performed. The immune cell types raised by in the epididymis will be analyzed by flow cytometry. The activation of mononuclear phagocytes will be evaluated by double immunofluorescence of clear cells and mononuclear phagocytes, and analyzed under high-resolution microscopy. All data will be statistically analyzed. (AU)