Human Cytomegalovirus Modifies Placental Small Extracellular Vesicle Composition to Enhance Infection of Fetal Neural Cells In Vitro.

Although placental small extracellular vesicles (sEVs) are extensively studied in the context of pregnancy, little is known about their role during viral congenital infection, especially at the beginning of pregnancy. In this study, we examined the consequences of human cytomegalovirus (hCMV) infection on sEVs production, composition, and function using an immortalized human cytotrophoblast cell line derived from first trimester placenta. By combining complementary approaches of biochemistry, electron microscopy, and quantitative proteomic analysis, we showed that hCMV infection increases the yield of sEVs produced by cytotrophoblasts and modifies their protein content towards a potential proviral phenotype. We further demonstrate that sEVs secreted by hCMV-infected cytotrophoblasts potentiate infection in naive recipient cells of fetal origin, including human neural stem cells. Importantly, these functional consequences are also observed with sEVs prepared from an ex vivo model of infected histocultures from early placenta. Based on these findings, we propose that placental sEVs could be important actors favoring viral dissemination to the fetal brain during hCMV congenital infection.

Usutu Virus Infects Human Placental Explants and Induces Congenital Defects in Mice.

Usutu virus (USUV) is a neurotropic mosquito-borne flavivirus that has dispersed quickly in Europe these past years. This arbovirus mainly follows an enzootic cycle involving mosquitoes and birds, but can also infect other mammals, causing notably sporadic cases in humans. Although it is mainly asymptomatic or responsible for mild clinical symptoms, USUV has been associated with neurological disorders, such as encephalitis and meningoencephalitis, highlighting the potential health threat of this virus. Among the different transmission routes described for other flaviviruses, the capacity for some of them to be transmitted vertically has been demonstrated, notably for Zika virus or West Nile virus, which are closely related to USUV. To evaluate the ability of USUV to replicate in the placenta and gain access to the fetus, we combined the use of several trophoblast model cell lines, ex vivo human placental explant cultures from first and third trimester of pregnancy, and in vivo USUV-infected pregnant mice. Our data demonstrate that human placental cells and tissues are permissive to USUV replication, and suggest that viral transmission can occur in mice during gestation. Hence, our observations suggest that USUV could be efficiently transmitted by the vertical route.

Effects of Acute Dengue Infection on Sperm and Virus Clearance in Body Fluids of Men.

We investigated the effects of dengue virus (DENV) on semen using samples collected 7, 15, 30, 60, and 90 days after symptom onset from 10 infected volunteers on Réunion Island. We assessed characteristics of semen and reproductive hormones and isolated motile spermatozoa from semen. We assayed semen for DENV using reverse transcription PCR and searched for DENV RNA by virus isolation in Vero E6 cell cultures. Four volunteers had >1 DENV RNA-positive semen samples; 2 volunteers had DENV RNA-positive semen at day 15 and 1 at day 30. No motile sperm were DENV positive. After exposure to positive semen, few Vero E6 cells stained positive for DENV antigens, indicating low levels of replicative virus. We found DENV had shorter duration in semen than in blood. These findings support the possibilities that DENV is sexually transmissible for a short period after acute dengue illness and that acute dengue induces reversible alterations in sperm.

Human Cytomegalovirus Infection Changes the Pattern of Surface Markers of Small Extracellular Vesicles Isolated From First Trimester Placental Long-Term Histocultures.

Extracellular vesicles (EVs) have increasingly been recognized as key players in a wide variety of physiological and pathological contexts, including during pregnancy. Notably, EVs appear both as possible biomarkers and as mediators involved in the communication of the placenta with the maternal and fetal sides. A better understanding of the physiological and pathological roles of EVs strongly depends on the development of adequate and reliable study models, specifically at the beginning of pregnancy where many adverse pregnancy outcomes have their origin. In this study, we describe the isolation of small EVs from a histoculture model of first trimester placental explants in normal conditions as well as upon infection by human cytomegalovirus. Using bead-based multiplex cytometry and electron microscopy combined with biochemical approaches, we characterized these small EVs and defined their associated markers and ultrastructure. We observed that infection led to changes in the expression level of several surface markers, without affecting the secretion and integrity of small EVs. Our findings lay the foundation for studying the functional role of EVs during early pregnancy, along with the identification of new predictive biomarkers for the severity and outcome of this congenital infection, which are still sorely lacking.

Anti-sperm antibodies detection by a modified MAR test: Towards a better definition of its indications.

RESEARCH QUESTION: Anti-sperm antibodies (ASA) have been shown to reduce male fertility but consensus about the precise situations in which tests should be carried out are lacking. In infertility investigations, should the mixed antiglobulin reaction (MAR) test be a first-line test? Should it be carried out systematically before assisted reproductive technology (ART)? What are the risk factors for ASA? DESIGN: All infertile patients (n = 1364) were tested with SpermMar (modified MAR test) between July 2013 and June 2017. Intra-patient variability of the MAR test was also assesed by comparing two tests within the same year in selected patients (n = 101). RESULTS: The main factor that influenced the percentage of ASA was the presence or absence of sperm agglutination. In the presence of agglutinations, 27 out of 72 (37.5%) patients were positive for ASA compared with 33 out of 1292 (2.6%) in the absence of agglutinations (P < 0.0001). When one risk factor was present (spontaneous sperm agglutination, history of scrotal trauma or inguinal surgery), 33 out of 179 (18.44%) tests were positive for ASA (≥50% coated spermatozoa), whereas only 27 out of 1242 (2.2%) were positive when no risk factor was present (P < 0.0001). CONCLUSIONS: ASA detection should not be systematically recommended in investigations of fertility status and before ART but reserved for when sperm agglutination is found during conventional sperm examination, or if the patient has a history of scrotal trauma or has undergone inguinal surgery.

Patterns of residual HIV-1 RNA shedding in the seminal plasma of patients on effective antiretroviral therapy.

BACKGROUND: More and more HIV-1-infected men on effective antiretroviral treatment (ART) have unprotected sex in order to procreate. The main factor influencing transmission is seminal HIV shedding. While the risk of HIV transmission is very low, it is difficult to assess in individuals. Nevertheless, it should be quantified. RESULTS: We retrospectively analysed seminal plasma HIV-1 shedding by 362 treated HIV-infected men attending a medically assisted reproduction centre (1998-2013) in order to determine its frequency, the impact of the antiretroviral regimen on HIV shedding, and to identify shedding patterns. The HIV-1 virus loads in 1396 synchronized blood and semen samples were measured, and antiretroviral treatment, biological and epidemiological data were recorded. We detected isolated HIV-1 shedding into the seminal plasma in 5.3% of patients on efficient antiretroviral treatment, but there was no association with the HIV antiretroviral drug regimen or the CD4 cell count. These men had undergone more regimen changes since treatment initiation and had been on the ongoing drug regimen longer than the non-shedding men. The patterns of HIV seminal shedding among patients with undetectable HIV blood virus load varied greatly. HIV seminal shedding can occur as long as 5 years after starting antiretroviral treatment. CONCLUSIONS: The seminal HIV load was used to monitor risk for infertile HIV-infected patients on an assisted reproductive technology program. This can still be recommended for patients who recently (6 months) started ART, or those with a poor history of adherence to ART but may also be usefull for some patients during counselling. Residual HIV seminal shedding is probably linked to breaks in adherence to antiretroviral treatment but local genital factors cannot be ruled out.

RÉSUMÉ: De plus en plus d'hommes sous traitement antirétroviral (ART) ont des rapports sexuels non protégés à des fins de procréation. Le principal déterminant de la transmission sexuelle est. l'excrétion séminale du VIH. Malgré un risque de transmission très faible, il reste difficile à évaluer au niveau individuel. Dans ce contexte, l'étude de l'excrétion séminale du VIH, notamment chez des hommes sous traitement antirétroviral, est. d'un grand intérêt. RÉSULTATS: Nous avons analysé rétrospectivement l'excrétion séminale du HIV chez 362 hommes sous traitement antirétroviral consultant un centre d'assistance médicale à la procréation (1998­2013) pour déterminer sa fréquence, l'impact des antirétroviraux sur l'excrétion du HIV et identifier les profils d'excrétion. Les charges virales HIV-1 ont été mesurées dans 1396 échantillons de sang et de sperme prélevés concomitamment et les traitements, les données biologiques et épidémiologiques recueillis. Nous avons détecté une excrétion dans le plasma séminal isolée chez 5,3% des patients sous traitement antirétroviral efficace mais nous n'avons pas trouvé d'association avec la composition du traitement antirétroviral ou le taux de lymphocytes T CD4+. Ces hommes avaient eu plus de changements thérapeutiques et leur traitement avait été instauré depuis plus longtemps que pour les hommes non excréteurs. Les profils d'excrétion séminale du HIV parmi les patients avec une charge virale indétectable dans le sang étaient très variables. L'excrétion séminale du HIV peut survenir jusqu'à 5 ans après l'instauration du premier traitement antirétroviral. CONCLUSIONS: La charge virale séminale du HIV était l'outil classique d'évaluation du risque viral de transmission pour les patients infertiles infectés par HIV et inclus dans les programmes d'assistance médicale à la procréation. Ceci peut continuer à être recommandé chez les patients ayant débuté un traitement antirétroviral dans les 6 mois précédent ou chez ceux avec des antécédents de mauvaise adhérence au traitement mais peut aussi être utile pour le conseil de certains patients. Le risque résiduel d'excrétion séminale du HIV est. probablement lié à des défauts d'adhérence au traitement antirétroviral mais des facteurs génitaux ne peuvent pas être éliminés.

Impact of Hodgkin or non-Hodgkin lymphoma and their treatments on sperm aneuploidy: a prospective study by the French CECOS network.

OBJECTIVE: To assess sperm production and aneuploidy in Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) before and after treatments. DESIGN: Multicenter, prospective, longitudinal study of lymphoma patients analyzed before treatment and after 3, 6, 12, and 24 months. SETTING: University hospitals. PATIENT(S): Forty-five HL and 13 NHL patients were investigated before and after treatment. Treatment regimens were classified in two groups: ABVD (doxorubicin, bleomycin, vinblastine, dacarbazine) with or without (±) radiotherapy, and CHOP (doxorubicin, cyclophosphamide, vincristine, prednisone)/MOPP-ABV (mechlorethamine, oncovin, procarbazine, prednisone-doxorubicin, bleomycin, vinblastine). A control group of 29 healthy men was also studied. INTERVENTION(S): Semen analyses and aneuploidy study by FISH were performed at each time point. MAIN OUTCOME MEASURE(S): Comparison of mean sperm characteristics and percentage of sperm aneuploidy rates before and after treatment. RESULT(S): Before treatment, HL and NHL men had altered semen characteristics and higher sperm aneuploidy rates (median 0.76 [interquartile range 0.56-0.64]) than the control group (0.54 [0.46-0.74]). After treatment, sperm production was significantly lowered 3 and 6 months after ABVD ± radiotherapy or CHOP/MOPP-ABV. After ABVD ± radiotherapy, the aneuploidy rate increased significantly only at 3 months, and values obtained 1 or 2 years later were lower than pretreatment values. In contrast, in the CHOP/MOPP-ABV treatment group, semen characteristics and aneuploidy rate did not return to normal levels until 2 years after treatment. CONCLUSION(S): Lymphoma itself has consequences on sperm aneuploidy frequency before treatment. Moreover, lymphoma treatments have deleterious effects on sperm chromosomes related to treatment type and time since treatment. Patient counseling is essential concerning the transient but significant sperm aneuploidy induced by lymphoma and its treatments.

Sperm DNA fragmentation after radioiodine treatment for differentiated thyroid cancer.

BACKGROUND: Treatment of differentiated thyroid cancer usually consists of a total thyroidectomy followed by one or several courses of radioiodine ((131)I). (131)I is known to have deleterious effects on radiation sensitive tissues and irradiation to the testes has been shown after its administration. We investigated effects of such treatment on sperm DNA in a patient with differentiated thyroid carcinoma. METHODS: The patient, a 32-year-old male with differentiated thyroid carcinoma treated by total thyroidectomy and radioiodine therapy, performed 6 semen samples in total, 3 for sperm banking and 3 for semen exploration, that were analysed for classic semen parameters. DNA integrity was analysed by flow cytometry: sperm DNA fragmentation index (DFI) and high DNA stainability (HDS) were analyzed by sperm chromatin structure assay, DNA fragmentation was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling assay. RESULTS: Moderate oligozoospermia was observed as early as 3 months after a first dose of (131)I and became severe at 5 months. Total sperm count was reduced up to 12 months after the second dose of (131)I. Sperm DFI was increased 3.25 months after the first dose of (131)I. All parameters returned to normal values 28 months after the second (131)I dose. CONCLUSIONS: Treatment with (131)I induces alterations in sperm chromatin as well as in sperm parameters a short time (3 months) after a first dose of (131)I with persistence of sperm alterations until 12 months after a second dose. Sperm banking should be recommended before treatment.

CONTEXTE: Le traitement du cancer différencié de la thyroïde consiste en général en une thyroïdectomie totale suivie d'un traitement par l'iode radioactif (131I). L'131I est connu pour ses effets délétères sur les tissus sensibles aux radiations et il a également été démontré une irradiation des testicules après administration d'iode radioactif. Nous avons donc cherché à savoir quels étaient les effets du traitement par l'iode radioactif sur l'ADN des spermatozoïdes. MÉTHODES: Le patient est un homme de 32 ans présentant un cancer différencié de la thyroïde traité par thyroïdectomie totale puis traitement par iodothérapie. Il a réalisé 6 prélèvements de sperme, 3 pour de l'autoconservation et 3 pour le suivi après traitement, dont les caractéristiques spermatiques ont été étudiées suivant les méthodes classiques. L'analyse de l'intégrité de la chromatine a été réalisée par cytométrie en flux : l'index de fragmentation de l'ADN (DFI) et la haute colorabilité de l'ADN (HDS) ont été analysés par le sperm chromatin structure assay (SCSA), la fragmentation de l'ADN a été évaluée par la technique de terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). RÉSULTATS: Une oligozoospermie modérée a été observée dès 3 mois après une première dose de 131I, et est devenue sévère à 5 mois. La numération des spermatozoïdes est restée réduite jusqu'à 12 mois après la seconde dose de 131I. Le DFI est élevé 3.25 mois après la première dose de 131I. Tous les paramètres ont retrouvé des valeurs normales 28 mois après la seconde dose d'131I. CONCLUSION: Le traitement par 131I induit des altérations de la chromatine du spermatozoïde en plus de celles des caractéristiques spermatiques une courte période (3mois) après une première dose de 131I, avec persistance de ces altérations spermatiques jusqu'à 12 mois après une seconde dose. L'autoconservation devrait être recommandée avant un tel traitement. MOTS CLÉ: Traitement par iode radioactif, SCSA, fragmentation de l'ADN du spermatozoïde, cancer différencié de la thyroïde, paramètres spermatiques.

Impact of lymphoma treatments on spermatogenesis and sperm deoxyribonucleic acid: a multicenter prospective study from the CECOS network.

OBJECTIVE: To determine consequences of lymphoma treatments on sperm characteristics and sperm DNA, and to evaluate predictors of sperm recovery. DESIGN: Multicenter prospective longitudinal study of patients analyzed before treatment and after 3, 6, 12, and 24 months. SETTING: University hospitals. PATIENT(S): Seventy-five Hodgkin lymphoma and non-Hodgkin lymphoma patients and a control group of 257 fertile men. INTERVENTION(S): Semen analyses, and sperm DNA and chromatin assessments. MAIN OUTCOME MEASURE(S): Comparisons of sperm characteristics before and after treatment. RESULT(S): Patients already had altered sperm characteristics before lymphoma treatment, with no identified risk factor. Sperm count, total sperm count, motility, and vitality decreased after treatment, with lowest values at 3 and 6 months. Twelve months after treatment, mean sperm count recovered to pretreatment values after doxorubicin, bleomycin, vinblastine, darcarbacine (ABVD) or ABVD+radiotherapy, but not after doxorubicin, cyclophosphamide, vincristine, prednisone (CHOP) or mechlorethamine, oncovin, procarbazine, prednisone (MOPP) chemotherapies. It was noteworthy that 7% of patients remained azoospermic at 24 months. After 24 months, Kaplan-Meier estimates showed that more than 90% of patients will recover normal sperm count after ABVD or ABVD+radiotherapy vs. 61% for CHOP chemotherapies. In multivariate analyses including diagnosis and treatment protocol, only pretreatment total sperm count was related to recovery. Compared with a control group, lymphoma patients had higher sperm chromatin alterations and DNA fragmentation before any treatment. After treatment, DNA fragmentation assessed by TUNEL assay and sperm chromatin structure assay decreased from 3 and 6 months, respectively, while remaining higher than in the control group during follow-up. CONCLUSION(S): Lymphoma patients had altered sperm DNA and chromatin before treatment. Lymphoma treatment had damaging effects on spermatogenesis. These data on both the recovery period according to treatment modalities and the pre- and post-treatment chromatin status of sperm are useful tools for counseling patients wishing to conceive.

Impact of chemotherapy and radiotherapy for testicular germ cell tumors on spermatogenesis and sperm DNA: a multicenter prospective study from the CECOS network.

OBJECTIVE: To determine the consequences of adjuvant testicular germ cell tumor treatment (TGCT) on sperm characteristics and sperm DNA, and to evaluate the predictors of sperm recovery. DESIGN: Multicenter prospective longitudinal study of patients analyzed before treatment and after 3, 6, 12, and 24 months. SETTING: University hospitals. PATIENT(S): One hundred twenty-nine volunteer TGCT patients and a control group of 257 fertile men. INTERVENTION(S): Routine semen analyses, sperm DNA, and chromatin assessments. MAIN OUTCOME MEASURE(S): Comparisons of mean sperm characteristics before and after treatment, with sperm recovery analyzed by the Kaplan-Meier method. RESULT(S): The quantitative and qualitative sperm characteristics decreased after treatment, with lowest values at 3 and 6 months and with variations according to treatment type. The mean total sperm count recovered to pretreatment values at 12 months after treatment after two or fewer bleomycin, etoposide, and cisplatin (BEP) cycles, but not after radiotherapy or more than two BEP cycles. Only the treatment modalities and pretreatment sperm production were related to recovery of the World Health Organization reference sperm values. An increased proportion of patients had elevated high sperm DNA stainability at 6 months after radiotherapy. CONCLUSION(S): Adjuvant treatments for testicular germ cell tumor have drastic effects on spermatogenesis and sperm chromatin quality. These new data on both the recovery period according to treatment modalities and the post-treatment chromatin status of sperm are useful tools for counseling patients wishing to conceive.

Antiviral effect of maraviroc in semen: a case report.

We describe the antiviral effect of maraviroc in a patient who had been shedding high levels of HIV-1 in seminal fluid for three years despite an undetectable blood plasma viral load. Adding maraviroc to HAART stopped the seminal shedding. We discuss the mechanisms involved and the effect on sexual transmission.

Mild induced testicular and epididymal hyperthermia alters sperm chromatin integrity in men.

OBJECTIVE: To investigate the effects of a mild induced testicular and epididymal hyperthermia (+2°C) on sperm chromatin integrity in men. DESIGN: Experimental prospective study. SETTING: University hospital. PATIENT(S): Five healthy fertile volunteers. INTERVENTION(S): Testicular and epididymal hyperthermia was induced by maintaining the testes at inguinal position with the support of specially designed underwear 15 ± 1 hours daily for 120 consecutive days. MAIN OUTCOME MEASURE(S): Classic semen characteristics. Sperm DNA fragmentation index (DFI) and high DNA stainability (HDS) were analyzed by sperm chromatin structure assay. RESULT(S): Compared with baseline values, sperm DFI and HDS were significantly increased as early as day (D) 20 and D34, respectively, and remained elevated during the entire period of hyperthermia. Percentages of motile and viable spermatozoa decreased as early as D20 and D34, respectively, and total sperm count decreased at D34 during hyperthermia and remained low during the entire hyperthermia period. All studied parameters returned to respective baseline values at D73 after cessation of hyperthermia. CONCLUSION(S): Mild induced testicular and epididymal hyperthermia largely impaired sperm chromatin integrity, which appeared before any changes in sperm output. These findings may have clinical implications in male contraception, infertility, and assisted reproductive technology.

Decrease of mitochondrial DNA level in sperm from patients infected with human immunodeficiency virus-1 linked to nucleoside analogue reverse transcriptase inhibitors.

OBJECTIVE: To study sperm parameters in patients infected with human immunodeficiency virus (HIV)-1 and to analyze mitochondrial DNA (mtDNA) in sperm according to the HIV treatment. DESIGN: Observational study. SETTING: University-affiliated teaching hospital. PATIENT(S): Thirty-two patients infected with HIV-1 and 31 noninfected healthy men provided semen samples. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): After DNA extraction, mtDNA level was assessed using real-time polymerase chain reaction (PCR) (LightCycler) in whole semen and in selected spermatozoa from 90% density centrifugation gradients. For each sample, mtDNA and ß-globin gene sequences were amplified and PCR products were quantified. The mtDNA-to-ß-globin ratio expressed the number of mtDNA copies per cell. RESULT(S): Compared with the control group, several sperm parameters were altered in patients with HIV. The number of mtDNA copies per cell in whole semen was increased in HIV-infected patients (6.3±6.3 vs. 3.5±3.2). However, there was no statistically significant difference in mtDNA copy number in the spermatozoa obtained after density gradient centrifugation. The number of nucleoside analogue reverse transcriptase inhibitors (NRTI) taken by patients during treatment significantly influenced the mtDNA level in sperm (1 NRTI 7.6±8.1, 2 NRTIs 7.0±5.1, 3 NRTIs 3.2±2.1). CONCLUSION(S): Using a specific method to measure sperm mtDNA, we demonstrated a decrease of mtDNA copies in spermatozoa after use of NRTIs with known mitochondrial toxicity.

Determining seminal plasma human immunodeficiency virus type 1 load in the context of efficient highly active antiretroviral therapy.

The semen plasma virus load is measured to ensure the safety of sperm processing during medically assisted procreation (MAP) for couples with a human immunodeficiency virus type 1 (HIV-1)-infected man. A practical, automated protocol using the COBAS Ampliprep CAP/CTM kit in the COBAS TaqMan96 system was developed to measure the HIV-1 load in semen plasma samples. HIV-1 was detected in 13.4% of the semen samples processed at our MAP center. Of the eight patients having a detectable semen HIV-1 load, five had no detectable virus in their blood plasma. This highlights the residual risk of HIV-1 transmission during unprotected intercourse and raises the question of the possible consequences of ineffective highly active antiretroviral therapy in the genital tract.

Ribavirin and pegylated interferon treatment for hepatitis C was associated not only with semen alterations but also with sperm deoxyribonucleic acid fragmentation in humans.

OBJECTIVE: To report the effects of ribavirin plus pegylated interferon on semen parameters and sperm DNA integrity in a man given treatment for hepatitis C. DESIGN: Case report. SETTING: University-affiliated teaching hospital. INTERVENTION(S): None. PATIENT(S): A 37-year-old man given treatment with ribavirin and pegylated interferon for hepatitis C. MAIN OUTCOME MEASURE(S): Semen parameters (sperm count, motility, round cells), sperm protamination state measured by sperm chromatin structure assay, and sperm DNA fragmentation measured by terminal uridine nick-end labeling assay. RESULT(S): The percentage of progressive spermatozoa and the number of motile sperm per ejaculate decreased during treatment. The round cell/spermatozoa ratio, which reflects spermatogenic abnormality, increased from 2.6% +/- 1.4% to 23.6% +/- 13.0% during treatment and returned to baseline value 4 months later. Moreover, the sperm DNA fragmentation index (as measured by sperm chromatin structure assay) increased very markedly during treatment (from 14.5% before to 69.2% at 7 months of treatment) and remained elevated 8 months later. CONCLUSION(S): This study reports for the first time not only quantitative but also qualitative alterations of spermatogenesis with DNA packaging abnormalities and emphasizes the need for prospective clinical studies. While the results of other studies are awaited, the alterations that persisted 8 months after treatment argue for a longer contraception period after treatment discontinuation in men.

Azoospermic HIV-1 infected patients wishing to have children: proposed strategy to reduce HIV-1 transmission risk during sperm retrieval and intracytoplasmic sperm injection: Case Report.

BACKGROUND: To date, assisted reproductive technology (ART) with sperm washing is offered to serodiscordant couples with an human immunodeficiency virus-1 (HIV-1) infected male partner in order to have a child while reducing the risk of transmission to the woman. However, ART programmes are not possible if the man is azoospermic. We report here the first birth following intracytoplasmic sperm injection (ICSI) using frozen epididymal spermatozoa obtained after surgical sperm retrieval in a HIV-1 infected man with obstructive azoospermia. METHODS; Sperm obtained by micro-epididymal sperm aspiration was frozen after density gradient preparation and tested for HIV-RNA and DNA. ICSI with frozen sperm was performed. RESULTS: A twin pregnancy was obtained following ICSI. Two healthy girls were born. Maternal HIV-1 RNA and HIV-1 serology were negative during pregnancy and at delivery. CONCLUSIONS: This case report demonstrates that ART is possible in azoospermic HIV-1 infected men. On the basis of current knowledge, we propose a strategy to reduce HIV-1 transmission risk during sperm retrieval and ICSI in couples where the man is HIV-1 infected and azoospermic.

Good efficiency of intrauterine insemination programme for serodiscordant couples with HIV-1 infected male partner: a retrospective comparative study.

OBJECTIVE: Assisted reproduction techniques can minimize the risk of HIV female contamination when the male partner is HIV-infected. The aim of this study was to investigate the efficiency of sperm washing and intrauterine insemination (IUI) in these couples. STUDY DESIGN: Retrospective comparative study. Eighty-four HIV-1 serodicordant couples underwent 294 IUI. The control group was composed of 90 couples (320 IUI cycles) with donor sperm. Spermatozoa from HIV-1 infected male partner were prepared and tested for HIV-1 according to sperm washing method. Spermatozoa from HIV-1 and donor male were frozen before IUI. IUI were performed after ovarian stimulation. Main outcomes measures were pregnancy rate per cycle and baby take-home rate per couples. RESULTS: Although the pregnancy rate and baby take-home rate were higher in IUI with sperm washing than in IUI using donor sperm (18.0 versus 14.7 and 52.4 versus 41.1, respectively), the differences were not statistically significant. In serodiscordant couples, blood estradiol levels under ovarian stimulation and total motile sperm inseminated were a determining factor in achieving pregnancy. No female HIV-1 contamination occurred. CONCLUSION: This study demonstrates that sperm washing and IUI are highly effective in enabling serodiscordant couples with an HIV-1 infected male partner to have a child.