Early Sertoli cell gene expression regulates pathogenesis in response to Ebola virus

Abstract Ebola virus (EBOV) persists and actively replicates in the reproductive tract of up to 50% of male disease survivors six month after infection1. Persistent EBOV infections are usually asymptomatic and can be transmitted sexually, but the host and viral factors that mediate these infections have not been characterized2,3. We assessed viral replication kinetics and host responses of mouse testicular Leydig and Sertoli cells infected with wild-type EBOV Makona (wtEBOV) and collected samples up to 28 days post-infection. While maximum EBOV viral load was similar in both cell types, viral transcription and replication was delayed in Sertoli cells compared to Leydig cells. Using RNAseq analysis, we found that Sertoli cells responded to wtEBOV infection more rapidly and robustly than did Leydig cells, and that early differential gene expression was primarily associated with membrane function and cell cycle activity. Experiments conducted with an EBOV virus-like particle model (vlpEBOV) indicated that differences in Leydig and Sertoli cell response occurred less than one-hour post-treatment, and evidence indicated that Sertoli cells limit early EBOV entry and/or transcription. Based on our analysis, we discuss how the roles played by Sertoli cells in immune privilege and spermatogenesis may affect their initial and continued response to EBOV infection and facilitate asymptomatic persistence.