| spelling |
Gleide Fernandes de Avelarhttp://lattes.cnpq.br/2440697413164924Luiz Renato de FrançaNathalie Dejucq-rainsfordLuiz Renato de FrançaRossana Correa Netto de MeloRicardo Mello MarinhoSamyra Maria dos Santos Nassif LacerdaFernando Marcos dos Reishttp://lattes.cnpq.br/9468901780810601Raquel de Lima Leite Soares Alvarenga2023-05-17T17:07:41Z2023-05-17T17:07:41Z2016-07-29http://hdl.handle.net/1843/535010000-0003-1591-2734O sêmen representa o principal vetor de disseminação do vírus da imunodeficiência humana (HIV) no mundo. A origem das partículas virais livres e das células infectadas presentes no fluido seminal, bem como a natureza das interações entre o HIV e os espermatozoides e as demais células espermatogênicas, permanecem ainda muito pouco conhecidas. O fato de número crescente de estudos terem descrito a persistência de vírus no sêmen de até 30% de indivíduos submetidos a diferentes combinações de tratamentos antirretrovirais, sugere que o sistema genital masculino representa um santuário anatômico/farmacológico que impede a erradicação do vírus no organismo. Neste contexto, é necessário investigar a permissividade das células espermatogênicas ao HIV, bem como a contribuição das mesmas para a carga viral do sêmen. A proposta deste estudo foi a de se utilizar a cultura de células testiculares isoladas para estudar a natureza das interações entre as células germinativas testiculares e o HIV-1 in vitro. Foi utilizada também a linhagem de células germinativas derivadas de seminoma, as TCam-2. Estas células foram analisadas para diferentes etapas da infecção: entrada viral, síntese e integração de DNA viral, síntese de proteína viral e produção viral. Foi observado que a transcrição reversa e integração do HIV em células Tcam-2 ocorre quando a entrada é favorecida por um pseudovírus VSV-HIV de amplo tropismo e após 24 horas de exposição à cloroquina. Foi também demonstrada a produção de partículas virais pelas células Tcam-2. Não foi possível confirmar a re-infecção com os vírus produzidos. Confirmou-se a ligação do vírus HIV-1 às celulas germinativas testiculares. Foi possível sugerir a internalização das partículas p24, tanto para as células Tcam-2 quanto para células germinativas testiculares através do estudo de secções seriadas obtidas por microscopia confocal. Estes resultados, em conjunto, sugerem que o HIV-1 pode ser internalizado por células espermatogênicas em cultura, embora este evento seja raro. Os eventos posteriores (transcrição reversa, integração) não foram observados em células germinativas testiculares.Semen is the most important mechanism for spreading of the human immunodeficiency virus (HIV) infection in the world. The source of free viral particles and the infected cells present in the seminal fluid, as well as the nature of the interactions between HIV and other spermatozoa and spermatogenic cells are still barely known. The fact that increasing numbers of studies have described the persistence of virus in semen up to 30% of patients undergoing different combinations of antiretroviral treatments, suggests that the male genital system is an anatomical/pharmacological sanctuary that prevents the eradication of the virus in the body. In this context, it is necessary to investigate the permissiveness of spermatogenic cells to HIV, as well as the contribution of these cells to the viral load of semen. The purpose of this study was to use the culture of isolated testicular cells to study the nature of interactions between testicular germ cells and HIV-1 in vitro. It was also used the germ cell lineage derived from seminoma, the TCAM-2. These cells were analyzed for different phases of infection: viral entry, viral DNA synthesis and integration of viral protein synthesis and viral production. It was observed that the reverse transcription and integration of HIV to the TCAM-2 cells occur when the input is favored by a large tropism VSV-HIV pseudovirus and after 24 hours of exposure to chloroquine. It was also demonstrated the production of viral particles by TCAM-2 cells, however we were unable to confirm the re-infection with the virus produced. On the other hand, it was demonstrated the binding of the HIV-1 virus to testicular germ cells. The internalization of p24 particles to both TCAM-2 and testicular germ cells was evaluatedthrough the study of serial sections obtained by confocal microscopy. These results together suggest that HIV-1 can be internalized by spermatogenic cells in culture, although this is a rare event. Also, further studies have to be done in order to evaluate the subsequent events, for instance reverse transcription and integration, after the virus entrance in testicular germ cells.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorporUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em Biologia CelularUFMGBrasilICB - INSTITUTO DE CIÊNCIAS BIOLOGICAShttp://creativecommons.org/licenses/by-nd/3.0/pt/info:eu-repo/semantics/openAccessBiologia celularHIVSêmenCélulas germinativasSeminomaGenitália masculinaHIVsêmencélulas germinativasTCam-2sistema genital masculinohomenHIV-1 e células germinativas humanas: possíveis vias e mecanismos de infecçãoPotential for virus endogenization in humans through testicular germ cell infection: the case of HIVinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALRaquel de Lima Leite Soares Alvarenga Tese de Doutorado HIV-1 E CÉLULAS GERMINATIVAS HUMANAS POSSÍVEIS VIAS E MECANISMOS DE INFECÇÃO.pdfRaquel de Lima Leite Soares Alvarenga Tese de Doutorado HIV-1 E CÉLULAS GERMINATIVAS HUMANAS POSSÍVEIS VIAS E MECANISMOS DE INFECÇÃO.pdfapplication/pdf8902512https://repositorio.ufmg.br/bitstream/1843/53501/4/Raquel%20de%20Lima%20Leite%20Soares%20Alvarenga%20Tese%20de%20Doutorado%20HIV-1%20E%20C%c3%89LULAS%20GERMINATIVAS%20HUMANAS%20POSS%c3%8dVEIS%20VIAS%20E%20MECANISMOS%20DE%20INFEC%c3%87%c3%83O.pdfe8e667cb5f9945a219347a6e487c6213MD54LICENSElicense.txtlicense.txttext/plain; charset=utf-82118https://repositorio.ufmg.br/bitstream/1843/53501/5/license.txtcda590c95a0b51b4d15f60c9642ca272MD55CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8805https://repositorio.ufmg.br/bitstream/1843/53501/2/license_rdf00e5e6a57d5512d202d12cb48704dfd6MD521843/535012023-05-17 14:07:42.228oai:repositorio.ufmg.br:1843/53501TElDRU7Dh0EgREUgRElTVFJJQlVJw4fDg08gTsODTy1FWENMVVNJVkEgRE8gUkVQT1NJVMOTUklPIElOU1RJVFVDSU9OQUwgREEgVUZNRwoKQ29tIGEgYXByZXNlbnRhw6fDo28gZGVzdGEgbGljZW7Dp2EsIHZvY8OqIChvIGF1dG9yIChlcykgb3UgbyB0aXR1bGFyIGRvcyBkaXJlaXRvcyBkZSBhdXRvcikgY29uY2VkZSBhbyBSZXBvc2l0w7NyaW8gSW5zdGl0dWNpb25hbCBkYSBVRk1HIChSSS1VRk1HKSBvIGRpcmVpdG8gbsOjbyBleGNsdXNpdm8gZSBpcnJldm9nw6F2ZWwgZGUgcmVwcm9kdXppciBlL291IGRpc3RyaWJ1aXIgYSBzdWEgcHVibGljYcOnw6NvIChpbmNsdWluZG8gbyByZXN1bW8pIHBvciB0b2RvIG8gbXVuZG8gbm8gZm9ybWF0byBpbXByZXNzbyBlIGVsZXRyw7RuaWNvIGUgZW0gcXVhbHF1ZXIgbWVpbywgaW5jbHVpbmRvIG9zIGZvcm1hdG9zIMOhdWRpbyBvdSB2w61kZW8uCgpWb2PDqiBkZWNsYXJhIHF1ZSBjb25oZWNlIGEgcG9sw610aWNhIGRlIGNvcHlyaWdodCBkYSBlZGl0b3JhIGRvIHNldSBkb2N1bWVudG8gZSBxdWUgY29uaGVjZSBlIGFjZWl0YSBhcyBEaXJldHJpemVzIGRvIFJJLVVGTUcuCgpWb2PDqiBjb25jb3JkYSBxdWUgbyBSZXBvc2l0w7NyaW8gSW5zdGl0dWNpb25hbCBkYSBVRk1HIHBvZGUsIHNlbSBhbHRlcmFyIG8gY29udGXDumRvLCB0cmFuc3BvciBhIHN1YSBwdWJsaWNhw6fDo28gcGFyYSBxdWFscXVlciBtZWlvIG91IGZvcm1hdG8gcGFyYSBmaW5zIGRlIHByZXNlcnZhw6fDo28uCgpWb2PDqiB0YW1iw6ltIGNvbmNvcmRhIHF1ZSBvIFJlcG9zaXTDs3JpbyBJbnN0aXR1Y2lvbmFsIGRhIFVGTUcgcG9kZSBtYW50ZXIgbWFpcyBkZSB1bWEgY8OzcGlhIGRlIHN1YSBwdWJsaWNhw6fDo28gcGFyYSBmaW5zIGRlIHNlZ3VyYW7Dp2EsIGJhY2stdXAgZSBwcmVzZXJ2YcOnw6NvLgoKVm9jw6ogZGVjbGFyYSBxdWUgYSBzdWEgcHVibGljYcOnw6NvIMOpIG9yaWdpbmFsIGUgcXVlIHZvY8OqIHRlbSBvIHBvZGVyIGRlIGNvbmNlZGVyIG9zIGRpcmVpdG9zIGNvbnRpZG9zIG5lc3RhIGxpY2Vuw6dhLiBWb2PDqiB0YW1iw6ltIGRlY2xhcmEgcXVlIG8gZGVww7NzaXRvIGRlIHN1YSBwdWJsaWNhw6fDo28gbsOjbywgcXVlIHNlamEgZGUgc2V1IGNvbmhlY2ltZW50bywgaW5mcmluZ2UgZGlyZWl0b3MgYXV0b3JhaXMgZGUgbmluZ3XDqW0uCgpDYXNvIGEgc3VhIHB1YmxpY2HDp8OjbyBjb250ZW5oYSBtYXRlcmlhbCBxdWUgdm9jw6ogbsOjbyBwb3NzdWkgYSB0aXR1bGFyaWRhZGUgZG9zIGRpcmVpdG9zIGF1dG9yYWlzLCB2b2PDqiBkZWNsYXJhIHF1ZSBvYnRldmUgYSBwZXJtaXNzw6NvIGlycmVzdHJpdGEgZG8gZGV0ZW50b3IgZG9zIGRpcmVpdG9zIGF1dG9yYWlzIHBhcmEgY29uY2VkZXIgYW8gUmVwb3NpdMOzcmlvIEluc3RpdHVjaW9uYWwgZGEgVUZNRyBvcyBkaXJlaXRvcyBhcHJlc2VudGFkb3MgbmVzdGEgbGljZW7Dp2EsIGUgcXVlIGVzc2UgbWF0ZXJpYWwgZGUgcHJvcHJpZWRhZGUgZGUgdGVyY2Vpcm9zIGVzdMOhIGNsYXJhbWVudGUgaWRlbnRpZmljYWRvIGUgcmVjb25oZWNpZG8gbm8gdGV4dG8gb3Ugbm8gY29udGXDumRvIGRhIHB1YmxpY2HDp8OjbyBvcmEgZGVwb3NpdGFkYS4KCkNBU08gQSBQVUJMSUNBw4fDg08gT1JBIERFUE9TSVRBREEgVEVOSEEgU0lETyBSRVNVTFRBRE8gREUgVU0gUEFUUk9Dw41OSU8gT1UgQVBPSU8gREUgVU1BIEFHw4pOQ0lBIERFIEZPTUVOVE8gT1UgT1VUUk8gT1JHQU5JU01PLCBWT0PDiiBERUNMQVJBIFFVRSBSRVNQRUlUT1UgVE9ET1MgRSBRVUFJU1FVRVIgRElSRUlUT1MgREUgUkVWSVPDg08gQ09NTyBUQU1Cw4lNIEFTIERFTUFJUyBPQlJJR0HDh8OVRVMgRVhJR0lEQVMgUE9SIENPTlRSQVRPIE9VIEFDT1JETy4KCk8gUmVwb3NpdMOzcmlvIEluc3RpdHVjaW9uYWwgZGEgVUZNRyBzZSBjb21wcm9tZXRlIGEgaWRlbnRpZmljYXIgY2xhcmFtZW50ZSBvIHNldSBub21lKHMpIG91IG8ocykgbm9tZXMocykgZG8ocykgZGV0ZW50b3IoZXMpIGRvcyBkaXJlaXRvcyBhdXRvcmFpcyBkYSBwdWJsaWNhw6fDo28sIGUgbsOjbyBmYXLDoSBxdWFscXVlciBhbHRlcmHDp8OjbywgYWzDqW0gZGFxdWVsYXMgY29uY2VkaWRhcyBwb3IgZXN0YSBsaWNlbsOnYS4KRepositório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2023-05-17T17:07:42Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
|