Investigação da infecção por Zika vírus no sistema genital masculino de camundongo
Carregando...
Data
2019-03-01
Autores
Lima, Maria Letícia Duarte
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
O Zika virus (ZIKV), arbovírus pertencente à família Flaviviridae, gênero Flavivirus, teve seu primeiro caso de transmissão autóctone relatado no Brasil em 2015. Sua rápida dispersão, não só pelo Brasil, mas por diversos países do mundo, tem trazido muitas preocupações, principalmente por sua infecção estar relacionada com vários casos de síndrome congênita do Zika em fetos de gestantes infectadas, assim como o aumento do número de casos da síndrome de Guillain-Barré. Além da transmissão por vetores artrópodes, casos de transmissão sexual foram relatados e verificou-se altos títulos virais no sêmen, mesmo após o fim da viremia. Um estudo detectou o vírus no sêmen por 62 dias após o fim dos sintomas, enquanto outro verificou por mais de um ano a presença do vírus nesse mesmo fluido. Para melhor esclarecer a infecção por ZIKV no sistema genital masculino, seus efeitos e em quais órgãos desse sistema o vírus se replica, modelos animais vêm sendo utilizados para tal, dentre eles o camundongo AG129. Este trabalho teve como objetivo estudar a infecção por ZIKV em células e órgãos do sistema genital masculino desses camundongos no pico da viremia. Foram avaliados: testículos, epidídimo e complexo prostático (vesícula seminal e próstata) dois dias após a infecção com o vírus. Para isso, foram utilizados 11 camundongos AG129, divididos em 2 grupos por metodologia: 5 para PCR em tempo real e 6 para imuno-histoquímica. Utilizamos o sangue e cérebro como controle positivo e um camundongo não-infectado em cada metodologia com controle negativo. Detectamos, através de qPCR, a presença do vírus em todos os órgãos analisados e obtivemos a seguinte média de títulos virais (em cópias de RNA) em cada: 6,76x108 no cérebro; 7,6x108 nos testículos; 6,13x108 nos epidídimos; 4,41x107 no complexo prostático e 9,41x1010 no soro. As imuno-histoquímicas demonstraram uma infecção inicial nestes órgãos, em que o vírus pode ser encontrado por muitas vezes próximo ou dentro de vasos sanguíneos, sendo que também pode ter sido observada algumas marcações no tecido em si. Nos testículos e epidídimos a infecção nos tecidos se mostrou mais significativa do que na próstata, mas nesta última também foi possível observar a imunorreatividade para ZIKV em seus epitélios. A verificação da proteína viral não-estrutural NS5, a RNA polimerase viral, nos tecidos dos testículos, epidídimos e até mesmo da próstata dá indícios de que o vírus é capaz de se replicar nestes órgãos. Portanto, a análise em 2 dias após a infecção destes órgãos, demonstrou altos títulos de RNA viral e uma infecção inicial nos tecidos, sendo mais robustas em testículo e epidídimo, além da imunorreatividade próxima a vasos e em leucócitos no interior deles.
Zika virus (ZIKV), arbovirus belonging to the Flaviviridae family, genus Flavivirus, had its first case of autochthonous transmission reported in Brazil in 2015. Its rapid dispersion, not only by Brazil, but by several countries of the world, has brought many concerns, mainly due to its infection is related to many cases of Congenital Zika Syndrome in fetuses of infected pregnant women, as well as the increase in the number of cases of Guillain-Barré syndrome. In addition to transmission by arthropod vectors, cases of sexual transmission have been reported and high viral titers have been found in semen, even after the end of viremia. One study detected the virus in the semen for 62 days after the end of symptoms, while another verified for more than a year the presence of the virus in that same fluid. In order to better clarify the ZIKV infection in the male genital system, its effects and in which organs of this system the virus replicates, animal models have been used for this, among them the mouse AG129. This work aimed to study ZIKV infection in cells and organs of the male genital tract of these mice at peak of viremia. Testis, epididymis and prostatic complex (seminal vesicle and prostate) were evaluated two days after infection with the virus. For this, 11 AG129 mice were used, divided into 2 groups by methodology: 5 for real-time PCR and 6 for immunohistochemistry. We used the blood and brain as a positive control and an uninfected mouse in each negative control methodology. We detected, through qPCR, the presence of the virus in all analyzed organs and obtained the following mean viral titers (in RNA copies) in each: 6.76x108 in the brain; 7.6x108 in the testicles; 6,13x108 in the epididymides; 4.41x107 in the prostatic complex and 9.4x1010 in the serum. Immunohistochemistry has demonstrated an initial infection in these organs, in which the virus can be found often near or within blood vessels, and some markings on the tissue itself may also have been observed. In the testis and epididymis tissue infection was more significant than in the prostate, but in the latter it was also possible to observe the immunoreactivity to ZIKV in its epithelia. The verification of NS5 nonstructural viral protein, viral RNA polymerase, in the tissues of the testes, epididymis and even the prostate gives evidence that the virus is able to replicate in these organs. Therefore, the analysis at 2 days post-infection of these organs showed high titers of viral RNA and an initial infection in the tissues, being more robust in testicles and epididymis, besides the immunoreactivity close to vessels and in leukocytes inside them.
Zika virus (ZIKV), arbovirus belonging to the Flaviviridae family, genus Flavivirus, had its first case of autochthonous transmission reported in Brazil in 2015. Its rapid dispersion, not only by Brazil, but by several countries of the world, has brought many concerns, mainly due to its infection is related to many cases of Congenital Zika Syndrome in fetuses of infected pregnant women, as well as the increase in the number of cases of Guillain-Barré syndrome. In addition to transmission by arthropod vectors, cases of sexual transmission have been reported and high viral titers have been found in semen, even after the end of viremia. One study detected the virus in the semen for 62 days after the end of symptoms, while another verified for more than a year the presence of the virus in that same fluid. In order to better clarify the ZIKV infection in the male genital system, its effects and in which organs of this system the virus replicates, animal models have been used for this, among them the mouse AG129. This work aimed to study ZIKV infection in cells and organs of the male genital tract of these mice at peak of viremia. Testis, epididymis and prostatic complex (seminal vesicle and prostate) were evaluated two days after infection with the virus. For this, 11 AG129 mice were used, divided into 2 groups by methodology: 5 for real-time PCR and 6 for immunohistochemistry. We used the blood and brain as a positive control and an uninfected mouse in each negative control methodology. We detected, through qPCR, the presence of the virus in all analyzed organs and obtained the following mean viral titers (in RNA copies) in each: 6.76x108 in the brain; 7.6x108 in the testicles; 6,13x108 in the epididymides; 4.41x107 in the prostatic complex and 9.4x1010 in the serum. Immunohistochemistry has demonstrated an initial infection in these organs, in which the virus can be found often near or within blood vessels, and some markings on the tissue itself may also have been observed. In the testis and epididymis tissue infection was more significant than in the prostate, but in the latter it was also possible to observe the immunoreactivity to ZIKV in its epithelia. The verification of NS5 nonstructural viral protein, viral RNA polymerase, in the tissues of the testes, epididymis and even the prostate gives evidence that the virus is able to replicate in these organs. Therefore, the analysis at 2 days post-infection of these organs showed high titers of viral RNA and an initial infection in the tissues, being more robust in testicles and epididymis, besides the immunoreactivity close to vessels and in leukocytes inside them.
Descrição
Palavras-chave
ZIKV, Zika vírus, Sistema genital masculino, Camundongo, AG129, Male reproductive tract, Mice