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Viral replication analysis and RSK2 protein expression analysis in CD4+ t cells infected by HIV-1 under several activation stimuli

Grant number: 18/05184-6
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): June 01, 2018
Effective date (End): December 31, 2018
Field of knowledge:Biological Sciences - Microbiology
Principal Investigator:Luiz Mário Ramos Janini
Grantee:Tatiane Montes de Andrade
Home Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil

Abstract

Introduction: The eradication of HIV virus from infected patients is arduous since HIV is capable of establish viral reservoirs. When the integration of the viral genetic material is made at genome regions of host cells that transcription is inactive, viral products have not enough expression to be target by antiretroviral therapy and to be detected by host Immunology System. According to that, infection can succeed to a latent phase. It is know that viral production depends on: chromatin epigenetic changes and the activity of transcription factors related to HIV integrated regions. The transcription factor NF-ºB can link to LTR-HIV promoter and it can increase viral replication. That way, this project has as propose investigate whether NF-ºB activated by RSK2 protein of MAPK pathway can effect viral replication. Objective: to investigate viral production and RSK2 expression after to stimulate CD4+ T cells by several substances. Materials and methods: from a concentration of erythrocytes and leukocytes Peripheral Blood Mononuclear Cells (PBMCs) will be separated by density gradient using Ficoll Paque reagent. Those cells will be treated for obtaining CD4+ T cells that will be activated by several substances. They will be infected by HIV-1 virus and cultivated in suitable culture medium. Cell culture supernatants will be used to detect viral RNA and protein extracts will be used to verify and quantify some protein expressions.