Close contact through sexual activity has been associated with the spread of monkeypox virus (MPXV) in the ongoing, global 2022 epidemic. However, it remains unclear whether MPXV replicates in the testes or is transmitted via semen to produce an active infection. We carried out a retrospective analysis of MPXV-infected crab-eating macaque archival tissue samples from acute and convalescent phases of infection of clade I or clade II MPXV using immunostaining and RNA in situ hybridization. We detected MPXV in interstitial cells and seminiferous tubules of testes as well as epididymal lumina, which are the sites of sperm production and maturation. We also detected inflammation and necrosis during the acute phase of the disease by histological analysis. Finally, we found that MPXV was cleared from most organs during convalescence, including healed skin lesions, but could be detected for up to 37 d post-exposure in the testes of convalescent macaques. Our findings highlight the potential for sexual transmission of MPXV in humans.
This is a preview of subscription content, access via your institution
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 12 digital issues and online access to articles
$119.00 per year
only $9.92 per issue
Rent or buy this article
Prices vary by article type
Prices may be subject to local taxes which are calculated during checkout
Magnus, P. V., Andersen, E. K., Petersen, K. B. & Birch-Andersen, A. A pox-like disease in cynomolgus monkeys. Acta Pathol. Microbiol. Scand. 46, 156–176 (1959).
Breman, J. G. et al. Human monkeypox, 1970–79. Bull. World Health Organ. 58, 165–182 (1980).
Beer, E. M. & Rao, V. B. A systematic review of the epidemiology of human monkeypox outbreaks and implications for outbreak strategy. PLoS Negl. Trop. Dis. 13, e0007791 (2019).
Bunge, E. M. et al. The changing epidemiology of human monkeypox–A potential threat? A systematic review. PLoS Negl. Trop. Dis. 16, e0010141 (2022).
Sklenovska, N. & Van Ranst, M. Emergence of monkeypox as the most important orthopoxvirus infection in humans. Front. Public Health 6, 241 (2018).
Yinka-Ogunleye, A. et al. Outbreak of human monkeypox in Nigeria in 2017–18: a clinical and epidemiological report. Lancet Infect. Dis. 19, 872–879 (2019).
Happi, C. et al. Urgent need for a non-discriminatory and non-stigmatizing nomenclature for monkeypox virus. PLoS Biol. 20, e3001769 (2022).
Erez, N. et al. Diagnosis of imported monkeypox, Israel, 2018. Emerg. Infect. Dis. 25, 980–983 (2019).
Vaughan, A. et al. Human-to-human transmission of monkeypox virus, United Kingdom, October 2018. Emerg. Infect. Dis. 26, 782–785 (2020).
Hobson, G. et al. Family cluster of three cases of monkeypox imported from Nigeria to the United Kingdom, May 2021. Eur. Surveill. 26, 2100745 (2021).
Ng, O. T. et al. A case of imported monkeypox in Singapore. Lancet Infect. Dis. 19, 1166 (2019).
Costello, V. et al. Imported monkeypox from international traveler, Maryland, USA, 2021. Emerg. Infect. Dis. 28, 1002–1005 (2022).
Rao, A. K. et al. Monkeypox in a traveler returning from Nigeria - Dallas, Texas, July 2021. Morb. Mortal. Wkly Rep. 71, 509–516 (2022).
Reed, K. D. et al. The detection of monkeypox in humans in the Western Hemisphere. N. Engl. J. Med. 350, 342–350 (2004).
Centers for Disease Control and Prevention. Update: multistate outbreak of monkeypox–Illinois, Indiana, Kansas, Missouri, Ohio, and Wisconsin, 2003. Morb. Mortal. Wkly Rep. 52, 642–646 (2003).
Vivancos, R. et al. Community transmission of monkeypox in the United Kingdom, April to May 2022. Eurosurveillance. 27, 2200422 (2022).
Minhaj, F. S. et al. Monkeypox outbreak - Nine States, May 2022. Morb. Mortal. Wkly Rep. 71, 764–769 (2022).
WHO Director-General’s Statement at the Press Conference Following IHR Emergency Committee Regarding The Multi-country Outbreak of Monkeypox (WHO, 2022).
WHO Health Emergency Dashboard-Monkeypox (WHO, 2022).
Monkeypox Key Facts (WHO, 2022).
Miura, F. et al. Estimated incubation period for monkeypox cases confirmed in the Netherlands, May 2022. Eurosurveillance 27, 2200448 (2022).
Perez Duque, M. et al. Ongoing monkeypox virus outbreak, Portugal, 29 April to 23 May 2022. Eurosurveillance 27, 2200424 (2022).
Antinori, A. et al. Epidemiological, clinical and virological characteristics of four cases of monkeypox support transmission through sexual contact, Italy, May 2022. Eurosurveillance 27, 2200421 (2022).
Thornhill, J. P. et al. Monkeypox virus infection in humans across 16 Countries – April–June 2022. N. Engl. J. Med. 387, 679–691 (2022).
Rao, A. K. et al. Use of JYNNEOS (smallpox and monkeypox vaccine, live, nonreplicating) for preexposure vaccination of persons at risk for occupational exposure to orthopoxviruses: recommendations of the Advisory Committee on Immunization Practices - United States, 2022. Morb. Mortal. Wkly Rep. 71, 734–742 (2022).
Tecovirimat SIGA (European Medicines Agency, 2022); https://www.ema.europa.eu/en/medicines/human/EPAR/tecovirimat-siga
Khodakevich, L., Jezek, Z. & Kinzanzka, K. Isolation of monkeypox virus from wild squirrel infected in nature. Lancet 1, 98–99 (1986).
Radonic, A. et al. Fatal monkeypox in wild-living sooty mangabey, Cote d’Ivoire, 2012. Emerg. Infect. Dis. 20, 1009–1011 (2014).
Doty, J. B. et al. Assessing monkeypox virus prevalence in small mammals at the human-animal interface in the Democratic Republic of the Congo. Viruses 9, 283 (2017).
Monkeypox (Centers for Disease Control and Prevention, 2022); https://www.cdc.gov/poxvirus/monkeypox/transmission.html
Johnson, R. F. et al. Comparative analysis of monkeypox virus infection of cynomolgus macaques by the intravenous or intrabronchial inoculation route. J. Virol. 85, 2112–2125 (2011).
Zaucha, G. M., Jahrling, P. B., Geisbert, T. W., Swearengen, J. R. & Hensley, L. The pathology of experimental aerosolized monkeypox virus infection in cynomolgus monkeys (Macaca fascicularis). Lab. Invest. 81, 1581–1600 (2001).
Huggins, J. et al. Nonhuman primates are protected from smallpox virus or monkeypox virus challenges by the antiviral drug ST-246. Antimicrob. Agents Chemother. 53, 2620–2625 (2009).
Nalca, A. et al. Experimental infection of cynomolgus macaques (Macaca fascicularis) with aerosolized monkeypox virus. PLoS ONE 5, e12880 (2010).
Goff, A. J. et al. A novel respiratory model of infection with monkeypox virus in cynomolgus macaques. J. Virol. 85, 4898–4909 (2011).
Chapman, J. L., Nichols, D. K., Martinez, M. J. & Raymond, J. W. Animal models of orthopoxvirus infection. Vet. Pathol. 47, 852–870 (2010).
Jordan, R. et al. ST-246 antiviral efficacy in a nonhuman primate monkeypox model: determination of the minimal effective dose and human dose justification. Antimicrob. Agents Chemother. 53, 1817–1822 (2009).
Shchelkunov, S. N. et al. Human monkeypox and smallpox viruses: genomic comparison. FEBS Lett. 509, 66–70 (2001).
Coffin, K. M. et al. Persistent Marburg virus infection in the testes of nonhuman primate survivors. Cell Host Microbe 24, 405–416.e3 (2018).
Zeng, X. et al. Identification and pathological characterization of persistent asymptomatic Ebola virus infection in rhesus monkeys. Nat. Microbiol. 2, 17113 (2017).
Smith, D. R. et al. Persistent Crimean-Congo hemorrhagic fever virus infection in the testes and within granulomas of non-human primates with latent tuberculosis. PLoS Pathog. 15, e1008050 (2019).
Liu, J. et al. Ebola virus persistence and disease recrudescence in the brains of antibody-treated nonhuman primate survivors. Sci. Transl. Med. 14, eabi5229 (2022).
Liu, J. et al. Nipah virus persists in the brains of nonhuman primate survivors. JCI Insight 4, e129629 (2019).
Govero, J. et al. Zika virus infection damages the testes in mice. Nature 540, 438–442 (2016).
Perry, D. L. et al. Ebola virus localization in the macaque reproductive tract during acute Ebola virus disease. Am. J. Pathol. 188, 550–558 (2018).
Salam, A. P. & Horby, P. W. The breadth of viruses in human semen. Emerg. Infect. Dis. 23, 1922–1924 (2017).
Pshenichnaya, N. Y., Sydenko, I. S., Klinovaya, E. P., Romanova, E. B. & Zhuravlev, A. S. Possible sexual transmission of Crimean-Congo hemorrhagic fever. Int. J. Infect. Dis. 45, 109–111 (2016).
Griffin, D. E. Why does viral RNA sometimes persist after recovery from acute infections? PLoS Biol. 20, e3001687 (2022).
Heskin, J. et al. Transmission of monkeypox virus through sexual contact – a novel route of infection. J. Infect. (2022).
Peiró-Mestres, A. et al. Frequent detection of monkeypox virus DNA in saliva, semen, and other clinical samples from 12 patients, Barcelona, Spain, May to June 2022. Eurosurveillance 27, 2200503 (2022).
Lapa, D. et al. Monkeypox virus isolation from a semen sample collected in the early phase of infection in a patient with prolonged seminal viral shedding. Lancet Infect. Dis. 22, 1267–1269 (2022).
Deen, G. F. et al. Ebola RNA persistence in semen of Ebola virus disease survivors - Final Report. N. Engl. J. Med. 377, 1428–1437 (2017).
Isidro, J. et al. Phylogenomic characterization and signs of microevolution in the 2022 multi-country outbreak of monkeypoxvirus. Nat. Med. 28, 1569–1572 (2022).
We thank J. Writer (United States Army Medical Research Institute of Infectious Diseases (USAMRIID), Fort Detrick, Frederick, MD, USA) for critically editing the manuscript and J. Huggins (USAMRIID) for his contribution to the original animal studies. The views and conclusions contained in this document are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of the US Department of the Army, the US Department of Defense, or of the institutions and companies affiliated with the authors. In no event shall any of these entities have any responsibility or liability for any use, misuse, inability to use, or reliance upon the information contained herein. The US departments do not endorse any products or commercial services mentioned in this publication.
The authors declare no competing interests.
Peer review information
Nature Microbiology thanks Hideki Hasegawa and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Extended Data Fig. 1 Characterization of inflammatory cells in the testicular tissue with MPXV persistence.
a,b, Low magnification immunofluorescence imaging demonstrates infiltration of CD68+ monocytes/macrophages (magenta in a), CD3+ T cells (green in a), myeloperoxidase (MPO)+ neutrophil granulocytes (green in b), and CD8+ T cells (magenta in b) in the areas surrounding the granulomas in the testes of MPXV survivors. Debris of MPO+ neutrophil granulocytes (green in b) are concentrated in the necrotic center of granulomas (regions circled by white dashed lines) in which MPXV persists. Representative immunofluorescence staining image shown from n = 2 survivors. Nuclei were counterstained blue with 4′,6-diamidino-2-phenylindole (DAPI). Scale bar, 100 µm.
About this article
Cite this article
Liu, J., Mucker, E.M., Chapman, J.L. et al. Retrospective detection of monkeypox virus in the testes of nonhuman primate survivors. Nat Microbiol 7, 1980–1986 (2022). https://doi.org/10.1038/s41564-022-01259-w