Zika Virus Infects Human Sertoli Cells And Trespasses The Blood-Testes Barrier To Gain Entry Into The Seminiferous Epithelium

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2017-08

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Over the past decade, Zika virus (ZIKV) has re-emerged as a pathogen of major health concern in the Western Hemisphere. Specifically, since 2007, 76 countries have announced new outbreaks of ZIKV, 29 of which also report increased incidence of CNS malformations such as microcephaly and also Guillain-Barre Syndrome [1]. These alarming statistics combined with the unique ability of ZIKV to be transmitted both sexually and in utero highlight the urgent need to study the mechanisms of ZIKV pathogenesis in order to ultimately develop vaccines, effective anti-viral therapies, and policies to control the spread of ZIKV disease. While ZIKV can be detected in human seminal fluid for months after the clearance of viremia, the cellular targets and mechanisms associated with persistent infection in the testes remains unclear. Mouse and NHP studies have recently shown that ZIKV can infect and damage the seminiferous tubules within the testes [2-5]. The seminiferous tubules are an immune privileged organ with a tight blood-testes barrier also known as the Sertoli-cell barrier (SCB), which protects developing spermatozoa from peripheral pathogens and environmental toxins. However, increased inflammatory mediators such as TNF-α and IL-1β, matrix metalloproteinases, and cell-adhesion molecules (CAM) can disrupt the integrity of the SCB leading to pathologic outcomes [6, 7]. Therefore, the objective of this study was to characterize ZIKV replication kinetics and immune responses in primary human Sertoli cells (SC) and develop an in vitro SCB model to understand mechanisms of ZIKV transmigration across the barrier. We demonstrate that primary human SC are highly susceptible to ZIKV as compared to the closely related dengue virus and induced expression of IFN-α, key cytokines and celladhesion molecules (VCAM-1 and ICAM-1). Further, using an in vitro SCB model, we show that ZIKV was released on the adluminal side of the SCB model with higher efficiency when compared to the blood-brain barrier model. ZIKV-infected SC also exhibited enhanced adhesion of leukocytes that correlated with decrease in the SCB integrity. While ZIKV infection did not affect the expression of tight and adherens junction proteins such as ZO-1, claudin and JAM-A, exposure of SC to inflammatory mediators derived from ZIKV-infected macrophages led to the degradation of ZO-1 protein that correlated with increased SCB permeability. Collectively, our data suggest that infection of SC may be one of the crucial steps by which ZIKV gains access to the site of spermatozoa development and identifies SC as a therapeutic target. Finally, the SCB model opens up opportunities to assess interactions of SC with other testicular cells and lays the platform for future studies to test the ability of anti-ZIKV drugs to cross the barrier and clear testicular infection.

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