Semen impairment and occurrence of SARS-CoV-2 virus in semen after recovery from COVID-19

Background The presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in human semen and its role in virus contagion and semen quality after recovery from coronavirus disease 2019 (COVID-19) is still unclear. Recent reports evidence that, after SARS-CoV-2 infection, male reproductive function and semen quality may be damaged. Aim To evaluate the semen parameters and inflammation of sexually active men following recovery from SARS-CoV-2 infection at 1 month and 3 months follow-up after the second negative nasopharyngeal swab. Materials and methods A prospective cross-sectional study on sexually active men recovered from SARS-CoV-2 infection was performed. For previously hospitalized COVID-19 patients, data on serum inflammatory markers were retrospectively collected. One month after the second SARS-CoV-2 negative nasopharyngeal swab and 3 months later, four biological fluid samples, namely saliva, pre-ejaculation urine, semen, and post-ejaculation urine, were collected. The occurrence of SARS-CoV-2 RNA in the specimen was evaluated in all the biological fluids collected by RT-PCR. Female partners were retested if any specimen was found to be SARS-CoV-2 positive. Semen parameters were evaluated according to the World Health Organization manual edition V. Furthermore, semen inflammation was assessed by quantification of semen leukocytes and interleukin-8 (IL-8) levels and evaluation of a panel of sperm cytokine levels by a two-step ELISA method. Results A total of 43 men were enrolled in the study. Three patients (7%) tested positive for at least one sample (one saliva;one pre-ejaculation urine;one semen and one post-ejaculation urine), so the next day new nasopharyngeal swabs were collected. The results from these 3 patients and their partners were all negative for SARS-CoV-2. At 1-month follow-up, 25% of the men with recent SARS-Cov-2 infections and proven healing were oligo-cryptoazoospermic, despite the absence of virus RNA in semen. Of the 11 men with semen impairment, 8 were azoospermic and 3 were oligospermic. Serum inflammatory markers (procalcitonin and C-reactive protein) were analyzed in previously hospitalized patients both at admission and at peak of infection. Levels at admission were statistically significantly higher in patients resulting in crypto-azoospermic with respect to those resulting in normozoospermic (p = 0.05;p = 0.03 and p = 0.02, respectively) after healing. Oligo-crypto-azoospermia was significantly related to COVID-19 severity (P < 0.001). A total of 33 patients (76.7%) showed pathological levels of IL-8 in semen. Interleukin-1β and tumor necrosis factor-α levels were significantly negatively related to sperm total number and concentration, whereas interleukin-4 was correlated with sperm motility. At 3-months follow-up, 8/10 men with semen impairment showed an overall increase of semen parameters compared to levels assessed after 1 month. Of the 4 crypto-/azoo-spermic men 1 month after healing, 2 resulted oligozoospermic, 1 normozoospermic and only 1 remained azoospermic. Two of the 3 oligozoospermic men turned normozoozpermic. Semen cytokine levels remained elevated after 3 months, except for IL-6. Discussion and conclusion SARS-CoV-2 can be detected in saliva, urine, and semen in a small percentage of men who recovered from COVID-19. 25% of men who recovered from COVID-19 demonstrated oligo-crypto-azoospermia. Negative correlations between interleukin-1β and tumor necrosis factor-α and sperm number and the overall high levels of semen cytokines indicate a potential detrimental role of SARS-CoV-2 driven inflammation on spermatogenesis. An overall tendency to an improvement of semen parameters was found although a genital tract inflammatory condition appears to persist at least 3 months after COVID-19 recovery. Despite the low number of enrolled patients may limit the statistical power of study and the fact that the previous semen quality of these men was unknown, our results indicate that male of reproductive age recovering from COVID-19 deserve accurate follow-up for their fertility status.

Ex Vivo and In Vivo CD46 Receptor Utilization by Species D Human Adenovirus Serotype 26 (HAdV26)

Human adenovirus serotype 26 (Ad26) is used as a gene-based vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and HIV-1. However, its primary receptor portfolio remains controversial, potentially including sialic acid, coxsackie and adenovirus receptor (CAR), integrins, and CD46. We and others have shown that Ad26 can use CD46, but these observations were questioned on the basis of the inability to cocrystallize Ad26 fiber with CD46. Recent work demonstrated that Ad26 binds CD46 with its hexon protein rather than its fiber. We examined the functional consequences of Ad26 for infection in vitro and in vivo. Ectopic expression of human CD46 on Chinese hamster ovary cells increased Ad26 infection significantly. Deletion of the complement control protein domain CCP1 or CCP2 or the serine-threonine-proline (STP) region of CD46 reduced infection. Comparing wild-type and sialic acid-deficient CHO cells, we show that the usage of CD46 is independent of its sialylation status. Ad26 transduction was increased in CD46 transgenic mice after intramuscular (i.m.) injection but not after intranasal (i.n.) administration. Ad26 transduction was 10-fold lower than Ad5 transduction after intratumoral (i.t.) injection of CD46expressing tumors. Ad26 transduction of liver was 1,000-fold lower than that ofAd5 after intravenous (i.v.) injection. These data demonstrate the use of CD46 by Ad26 in certain situations but also show that the receptor has little consequence by other routes of administration. Finally, i.v. injection of high doses of Ad26 into CD46 mice induced release of liver enzymes into the bloodstream and reduced white blood cell counts but did not induce thrombocytopenia. This suggests that Ad26 virions do not induce direct clotting side effects seen during coronavirus disease 2019 (COVID-19) vaccination with this serotype of adenovirus. IMPORTANCE The human species D Ad26 is being investigated as a low-seroprevalence vector for oncolytic virotherapy and gene-based vaccination against HIV-1 and SARS-CoV-2. However, there is debate in the literature about its tropism and receptor utilization, which directly influence its efficiency for certain applications. This work was aimed at determining which receptor(s) this virus uses for infection and its role in virus biology, vaccine efficacy, and, importantly, vaccine safety.

Ex Vivo and In Vivo CD46 Receptor Utilization by Species D Human Adenovirus Serotype 26 (HAdV26).

Human adenovirus serotype 26 (Ad26) is used as a gene-based vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and HIV-1. However, its primary receptor portfolio remains controversial, potentially including sialic acid, coxsackie and adenovirus receptor (CAR), integrins, and CD46. We and others have shown that Ad26 can use CD46, but these observations were questioned on the basis of the inability to cocrystallize Ad26 fiber with CD46. Recent work demonstrated that Ad26 binds CD46 with its hexon protein rather than its fiber. We examined the functional consequences of Ad26 for infection in vitro and in vivo. Ectopic expression of human CD46 on Chinese hamster ovary cells increased Ad26 infection significantly. Deletion of the complement control protein domain CCP1 or CCP2 or the serine-threonine-proline (STP) region of CD46 reduced infection. Comparing wild-type and sialic acid-deficient CHO cells, we show that the usage of CD46 is independent of its sialylation status. Ad26 transduction was increased in CD46 transgenic mice after intramuscular (i.m.) injection but not after intranasal (i.n.) administration. Ad26 transduction was 10-fold lower than Ad5 transduction after intratumoral (i.t.) injection of CD46-expressing tumors. Ad26 transduction of liver was 1,000-fold lower than that ofAd5 after intravenous (i.v.) injection. These data demonstrate the use of CD46 by Ad26 in certain situations but also show that the receptor has little consequence by other routes of administration. Finally, i.v. injection of high doses of Ad26 into CD46 mice induced release of liver enzymes into the bloodstream and reduced white blood cell counts but did not induce thrombocytopenia. This suggests that Ad26 virions do not induce direct clotting side effects seen during coronavirus disease 2019 (COVID-19) vaccination with this serotype of adenovirus. IMPORTANCE The human species D Ad26 is being investigated as a low-seroprevalence vector for oncolytic virotherapy and gene-based vaccination against HIV-1 and SARS-CoV-2. However, there is debate in the literature about its tropism and receptor utilization, which directly influence its efficiency for certain applications. This work was aimed at determining which receptor(s) this virus uses for infection and its role in virus biology, vaccine efficacy, and, importantly, vaccine safety.

P0142 Semen impairment and occurrence of SARS-CoV-2 virus in semen after recovery from COVID-19

Introduction & Objectives: The presence of SARS-CoV-2 in human semen and its role in virus transmission and semen quality after recovery from coronavirus disease 2019 (COVID-19) is still undefined. To date, studies evaluating semen quality and the occurrence of SARS-CoV-2 in semen of infected or proven recovered men are scarce and include a limited number of cases. Aim of this study is to evaluate the semen quality of sexually active men following recovery from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Materials & Methods: A prospective cross-sectional study on sexually active men who were known to have recovered from SARS-CoV2 was performed, after Ethical Appraisal (ClinicalTrials.gov Identifier: NCT04446169). Four biological fluid samples, specifically saliva, pre-ejaculation urine, semen and post-ejaculation urine, were tested for the SARS-CoV-2 genome by RT-PCR. Routine semen parameters and quantification of semen leukocytes and interleukin-8 (IL-8) levels were evaluated according to the World Health Organization manual edition V and by a two-step ELISA method respectively. Questionnaires including International Index of Erectile Function and Male Sexual Health Questionnaire Short Form were administered to all subjects. Results: From 326SARS-CoV-2 positive male patients, 160 (49%) we re eligible.Amongthem,117were excluded, 55 because werenotreachable by phone, 46 refused to participate and other 16 were unable to collect semen samples. Therefore, 43 patients were finally enrolled. Among them. 3 patients (7%) tested positive for at least one sample (1 saliva;1 pre-ejaculation urine;1 semen and 1 post-ejaculation urine),so the day after new nasopharyngeal swabs were collected. The results from these three patients and their partners were all negative for SARS-CoV-2. Overall, 25% of the men studied were oligo-crypto-azoospermic after recovery from COVID-19. Of the 11 men with semen impairment, 8 were azoospermic and 3 were oligospermic. A total of 33 patients (76.7%) showed pathological levels of IL-8 in semen. Oligo-crypto-azoospermia was significantly related to COVID-19 severity (p<0.001). Conclusions: SARS-CoV-2 can be detected in saliva, urine and semen in a small percentage of men who recovered from COVID-19. One-quarter of men who recovered from COVID-19 demonstrated oligo-crypto-azoospermia showing that an evaluation of semen quality should be recommended for men of reproductive age who are affected by COVID-19.

Semen impairment and occurrence of SARS-CoV-2 virus in semen after recovery from COVID-19.

STUDY QUESTION: How is the semen quality of sexually active men following recovery from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection? SUMMARY ANSWER: Twenty-five percent of the men with recent SARS-Cov-2 infections and proven healing were oligo-crypto-azoospermic, despite the absence of virus RNA in semen. WHAT IS KNOWN ALREADY: The presence of SARS-CoV-2 in human semen and its role in virus contagion and semen quality after recovery from coronavirus disease 2019 (COVID-19) is still unclear. So far, studies evaluating semen quality and the occurrence of SARS-CoV-2 in semen of infected or proven recovered men are scarce and included a limited number of participants. STUDY DESIGN, SIZE, DURATION: A prospective cross-sectional study on 43 sexually active men who were known to have recovered from SARS-CoV2 was performed. Four biological fluid samples, namely saliva, pre-ejaculation urine, semen, and post-ejaculation urine, were tested for the SARS-CoV-2 genome. Female partners were retested if any specimen was found to be SARS-CoV-2 positive. Routine semen analysis and quantification of semen leukocytes and interleukin-8 (IL-8) levels were performed. PARTICIPANTS/MATERIALS, SETTING, METHODS: Questionnaires including International Index of Erectile Function and Male Sexual Health Questionnaire Short Form were administered to all subjects. The occurrence of virus RNA was evaluated in all the biological fluids collected by RT-PCR. Semen parameters were evaluated according to the World Health Organization manual edition V. Semen IL-8 levels were evaluated by a two-step ELISA method. MAIN RESULTS AND THE ROLE OF CHANCE: After recovery from COVID-19, 25% of the men studied were oligo-crypto-azoospermic. Of the 11 men with semen impairment, 8 were azoospermic and 3 were oligospermic. A total of 33 patients (76.7%) showed pathological levels of IL-8 in semen. Oligo-crypto-azoospermia was significantly related to COVID-19 severity (P < 0.001). Three patients (7%) tested positive for at least one sample (one saliva; one pre-ejaculation urine; one semen and one post-ejaculation urine), so the next day new nasopharyngeal swabs were collected. The results from these three patients and their partners were all negative for SARS-CoV-2. LIMITATIONS, REASONS FOR CAUTION: Although crypto-azoospermia was found in a high percentage of men who had recovered from COVID-19, clearly exceeding the percentage found in the general population, the previous semen quality of these men was unknown nor is it known whether a recovery of testicular function was occurring. The low number of enrolled patients may limit the statistical power of study. WIDER IMPLICATIONS OF THE FINDINGS: SARS-CoV-2 can be detected in saliva, urine, and semen in a small percentage of men who recovered from COVID-19. One-quarter of men who recovered from COVID-19 demonstrated oligo-crypto-azoospermia indicating that an assessment of semen quality should be recommended for men of reproductive age who are affected by COVID-19. STUDY FUNDING/COMPETING INTEREST(S): None. TRIAL REGISTRATION NUMBER: N/A.