TY - JOUR
T1 - High SARS-CoV-2 tropism and activation of immune cells in the testes of non-vaccinated deceased COVID-19 patients
AU - Costa, Guilherme M.J.
AU - Lacerda, Samyra M.S.N.
AU - Figueiredo, André F.A.
AU - Wnuk, Natália T.
AU - Brener, Marcos R.G.
AU - Andrade, Lídia M.
AU - Campolina-Silva, Gabriel H.
AU - Kauffmann-Zeh, Andrea
AU - Pacifico, Lucila G.G.
AU - Versiani, Alice F.
AU - Antunes, Maísa M.
AU - Souza, Fernanda R.
AU - Cassali, Geovanni D.
AU - Caldeira-Brant, André L.
AU - Chiarini-Garcia, Hélio
AU - de Souza, Fernanda G.
AU - Costa, Vivian V.
AU - da Fonseca, Flavio G.
AU - Nogueira, Maurício L.
AU - Campos, Guilherme R.F.
AU - Kangussu, Lucas M.
AU - Martins, Estefânia M.N.
AU - Antonio, Loudiana M.
AU - Bittar, Cintia
AU - Rahal, Paula
AU - Aguiar, Renato S.
AU - Mendes, Bárbara P.
AU - Procópio, Marcela S.
AU - Furtado, Thiago P.
AU - Guimaraes, Yuri L.
AU - Menezes, Gustavo B.
AU - Martinez-Marchal, Ana
AU - Orwig, Kyle E.
AU - Brieño-Enríquez, Miguel
AU - Furtado, Marcelo H.
N1 - Funding Information:
We thank Dr. Luiz Orlando Ladeira, Dr. Rodrigo Ribeiro Resende, Dr. Ary Correa Jr, and Dr. Iara Borges for their help and for gently allowing the use of their Lab equipment. We also thank the Image Acquisition and Processing Center (CAPI-ICB/UFMG), the BSL-3 Laboratory from the ICB-UFMG, and the Centro de Microscopia da UFMG (CM-UFMG) for their technical assistance. We especially thank the families of the deceased COVID-19 patients for understanding the importance of our study.
Funding Information:
This work was financed by Ferring COVID-19 Investigational Grant (grant n° FIN0042393, to G.M.J.C). G.M.J.C. also received resources from FAPEMIG (APQ-01078-21 and RED-00079-22) and PRPq-UFMG. Miguel A. Brieño-Enriquez and Ana Martinez-Marchal were supported by Eunice Kennedy Shriver National Institute of Child Health and Human Development (R00HD090289 and P50HD096723), Ferring COVID-19 Investigational Grants, MWRI Research & Education Funding and Magee Auxiliary Research Scholar (MARS) endowment. Other specific grants from coauthors punctually helped in some experiments and logistics of the present study. SMSNL was supported by FAPEMIG (RED-00135-22). AFV was supported by FAPESP (grant #18/17647-0) when the study was conducted. GRFC is supported by FAPESP (grant #20/07419-0). M.H.F. was supported by the Laboratório São Paulo/BH-MG. M.L.N. is supported by FAPESP (grant # 2019/07250-9 and #2020/04836-0) and CNPq. F.G.F. received grants from FAPEMIG (CBB-APQ-03081-17, and CBB- APQ-04295-17), and from Rede Mineira de Pesquisa e Inovação para Bioengenharia de Nanossistemas (RM PI-BEM – FAPEMIG - TEC - RED-00282-16). R.S.A was supported by CNPq (R.S.A.: 312688/2017-2 and 439119/2018-9).
Publisher Copyright:
© 2023, The Author(s).
PY - 2023/12
Y1 - 2023/12
N2 - Background: Cellular entry of SARS-CoV-2 has been shown to rely on angiotensin-converting enzyme 2 (ACE2) receptors, whose expression in the testis is among the highest in the body. Additionally, the risk of mortality seems higher among male COVID-19 patients, and though much has been published since the first cases of COVID-19, there remain unanswered questions regarding SARS-CoV-2 impact on testes and potential consequences for reproductive health. We investigated testicular alterations in non-vaccinated deceased COVID-19-patients, the precise location of the virus, its replicative activity, and the immune, vascular, and molecular fluctuations involved in the pathogenesis. Results: We found that SARS-CoV-2 testicular tropism is higher than previously thought and that reliable viral detection in the testis requires sensitive nanosensors or RT-qPCR using a specific methodology. Through an in vitro experiment exposing VERO cells to testicular macerates, we observed viral content in all samples, and the subgenomic RNA’s presence reinforced the replicative activity of SARS-CoV-2 in testes of the severe COVID-19 patients. The cellular structures and viral particles, observed by transmission electron microscopy, indicated that macrophages and spermatogonial cells are the main SARS-CoV-2 lodging sites, where new virions form inside the endoplasmic reticulum Golgi intermediate complex. Moreover, we showed infiltrative infected monocytes migrating into the testicular parenchyma. SARS-CoV-2 maintains its replicative and infective abilities long after the patient’s infection. Further, we demonstrated high levels of angiotensin II and activated immune cells in the testes of deceased patients. The infected testes show thickening of the tunica propria, germ cell apoptosis, Sertoli cell barrier loss, evident hemorrhage, angiogenesis, Leydig cell inhibition, inflammation, and fibrosis. Conclusions: Our findings indicate that high angiotensin II levels and activation of mast cells and macrophages may be critical for testicular pathogenesis. Importantly, our findings suggest that patients who become critically ill may exhibit severe alterations and harbor the active virus in the testes.
AB - Background: Cellular entry of SARS-CoV-2 has been shown to rely on angiotensin-converting enzyme 2 (ACE2) receptors, whose expression in the testis is among the highest in the body. Additionally, the risk of mortality seems higher among male COVID-19 patients, and though much has been published since the first cases of COVID-19, there remain unanswered questions regarding SARS-CoV-2 impact on testes and potential consequences for reproductive health. We investigated testicular alterations in non-vaccinated deceased COVID-19-patients, the precise location of the virus, its replicative activity, and the immune, vascular, and molecular fluctuations involved in the pathogenesis. Results: We found that SARS-CoV-2 testicular tropism is higher than previously thought and that reliable viral detection in the testis requires sensitive nanosensors or RT-qPCR using a specific methodology. Through an in vitro experiment exposing VERO cells to testicular macerates, we observed viral content in all samples, and the subgenomic RNA’s presence reinforced the replicative activity of SARS-CoV-2 in testes of the severe COVID-19 patients. The cellular structures and viral particles, observed by transmission electron microscopy, indicated that macrophages and spermatogonial cells are the main SARS-CoV-2 lodging sites, where new virions form inside the endoplasmic reticulum Golgi intermediate complex. Moreover, we showed infiltrative infected monocytes migrating into the testicular parenchyma. SARS-CoV-2 maintains its replicative and infective abilities long after the patient’s infection. Further, we demonstrated high levels of angiotensin II and activated immune cells in the testes of deceased patients. The infected testes show thickening of the tunica propria, germ cell apoptosis, Sertoli cell barrier loss, evident hemorrhage, angiogenesis, Leydig cell inhibition, inflammation, and fibrosis. Conclusions: Our findings indicate that high angiotensin II levels and activation of mast cells and macrophages may be critical for testicular pathogenesis. Importantly, our findings suggest that patients who become critically ill may exhibit severe alterations and harbor the active virus in the testes.
KW - Infertility
KW - Leydig cell
KW - Macrophages
KW - Nanotechnology
KW - Renin-angiotensin system
KW - SARS-CoV-2 replication
KW - Sertoli cell
KW - Spermatogenesis
KW - Spermatogonia
KW - Testosterone
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U2 - 10.1186/s12915-022-01497-8
DO - 10.1186/s12915-022-01497-8
M3 - Article
C2 - 36797789
AN - SCOPUS:85148252494
SN - 1741-7007
VL - 21
JO - BMC Biology
JF - BMC Biology
IS - 1
M1 - 36
ER -