Detection of Simian Immunodeficiency Virus in Semen, Urethra, and Male Reproductive Organs during Efficient Highly Active Antiretroviral Therapy

Abstract

A number of men receiving prolonged suppressive highly active antiretroviral therapy (HAART) still shed human immunodeficiency virus (HIV) in semen. To investigate whether this seminal shedding may be due to poor drug penetration and/or viral production by long-lived cells within male genital tissues, we analyzed semen and reproductive tissues from macaques chronically infected with simian immunodeficiency virus mac251 (SIVmac251) who were treated for 4 months with HAART, which was intensified over the last 7 weeks with an integrase inhibitor. We showed that a subset of treated animals continued shedding SIV in semen despite efficient HAART. This shedding was not associated with low antiretroviral drug concentrations in semen or in testis, epididymis, seminal vesicles, and prostate. HAART had no significant impact on SIV RNA in the urethra, whereas it drastically reduced SIV RNA levels in the prostate and vas deferens and to a lesser extent in the epididymis and seminal vesicle. The only detectable SIV RNA-positive cells within the male genital tract after HAART were urethral macrophages. SIV DNA levels in genital tissues were not decreased by HAART, suggesting the presence throughout the male genital tract of nonproductively infected cells. In conclusion, our results demonstrate that 4 months of HAART induced variable and limited control of viral infection in the male reproductive organs, particularly in the urethra, and suggest that infected long-lived cells in the male genital tract may be involved in persistent seminal shedding during HAART. These results pave the way for further investigations of male genital organ infection in long-term-treated infected individuals. IMPORTANCEA substantial subset of men receiving prolonged HAART suppressing viral loads in the blood still harbor HIV in semen, and cases of sexual transmission have been reported. To understand the origin of this persistence, we analyzed the semen and male reproductive tissues from SIV-infected macaques treated with HAART. We demonstrated that persistent seminal shedding was not linked to poor drug penetration in semen or semen-producing prostate, seminal vesicle, epididymis, and testis. We revealed that HAART decreased SIV RNA to various extents in all male genital organs, with the exception of the urethra, in which SIV RNA ؉ macrophages were observed despite HAART. Importantly, HAART did not impact SIV DNA levels in the male genital organs. These results suggest that infection of male genital organs, and particularly the urethra, could be involved in the release of virus in semen during HAART.A pproximately 35.3 million people worldwide are living with human immunodeficiency virus (HIV)/AIDS, and 2.3 million new infections occur annually (1). Over 80% of these new infections are caused by unprotected sexual intercourse. The risk of sexual transmission of HIV is strongly correlated with HIV RNA levels in genital secretions (2). Because semen is the most common vector of HIV dissemination worldwide (3-5), preventing HIV transmission by...

Zika virus infects human testicular tissue and germ cells

“…Tissues or cell pellets were fixed in 4% formaldehyde and embedded in paraffin. RNA ISH was performed using RNAscope 2.5 (Advanced Cell Diagnostics) according to the manufacturer's instructions, as previously described (71,72). RNA-Scope ISH is a highly specific and sensitive technique, with the ability to detect single molecules (72).…”

Tissue reservoirs of HIV

“…HIV-RNA can be detected in the semen in 2-48% of on-ART, aviremic men[115-119], sometimes in association with drug resistance mutations[115]. Similarly, a subset of ART-treated, SIV-infected macaques continued to shed SIV in semen despite suppression in the plasma[120] with greater reduction of SIV-RNA in prostate and vas deferens, smaller decreases in epididymis and seminal vesicle but no change in urethra due to SIV+ macrophages. Multiple studies implicate a local source of seminal plasma virus showing compartmentalization from blood[32,121-123], suggesting a distinct reservoir.…”

HIV-1 is undetectable in preejaculatory secretions from HIV-1-infected men on suppressive HAART

“…PE from HIV-infected men contains HIV-1-infected white blood cells (WBC) [25, 26], and delayed application of condoms is a risk factor for HIV-1 transmission among men who have sex with men (MSM), providing evidence that PE may be infectious [27, 28]. SIV-infected nonhuman primates, like men, harbor virus in the penile urethra [29], and a recent study in male macaques demonstrated that HAART does not reduce SIV RNA levels in the urethra [30]. The purpose of the present study was to determine whether HIV persists in pre-ejaculatory fluid from HIV-1 infected men on HAART, and whether it is associated with seminal HIV shedding.…”