Study On T Cell Transcription Profile Of HIV Infection And PYCARD On Virus Replication And Its Mechanism

Background: AIDS,also known as acquired immunodeficiency syndrome(AIDS),is caused by human immunodeficiency virus(HIV)infection.As a major disease that seriously threatens human health,it has always been the public health and social governance of various countries in the world.The major challenges in this regard have received extensive attention from scientists.Therefore,it is urgent to explore the characteristics of AIDS infection and its possible cure mechanism.This is not only in line with the national innovation-driven development strategy,but also closely related to national health and safety issues.HIV infection can cause changes in the function of T lymphocytes,leading to changes in the host's entire immune system and destruction of homeostasis.In addition,the characteristics of HIV infection vary with the period of infection,and these characteristics may determine the fate of patients;however,the specific pathways activated in different T cell subtypes at different stages of infection are still unclear.Purpose: HIV infection can cause changes in the function of T lymphocytes,leading to changes in the host's entire immune system and destruction of homeostasis.In addition,the characteristics of HIV infection vary with the period of infection,and these characteristics may determine the fate of patients;however,the specific pathways activated in different T cell subtypes at different stages of infection are still unclear.This study aims to clarify the transcriptional profile and metabolic characteristics of CD4+ T cells and CD8+ T cells after HIV infection.Discover key pathways,predict and screen proteins that can regulate HIV infection,and preliminarily verify its impact on HIV virus.Methods: By searching the Gene Expression Omnibus(GEO),the GEO data of early HIV infection,chronic progressive infection and uninfected persons were downloaded.The weighted gene co-expression network analysis(WGCNA)was used to evaluate the transcriptome characteristics of CD4+ and CD8+ T cells to find possible antiviral targets.A new metabolic algorithm was used to analyze the similarities and differences of the metabolic pathways of these two T cells at different stages of infection.Screen the genes of interest for the next step of verification,perform gene set enrichment analysis and protein interaction prediction,and verify through molecular biology experiments.Results: Weighted gene co-expression network analysis was used to evaluate changes in gene expression.We identified different functional modules.CD4+ T cells can be divided into 23 functional modules,and CD8+ T cells can be divided into 20 functional modules.Both have Different transcription characteristics.Through the level of correlation,we screened a key module at each stage of each cell,and then identified the module characteristics and key targets to determine the antiviral response.For example,early infection of CD4+ T cells is significantly enriched in metabolism and infection-related pathways,and chronic infection is significantly enriched in pathways such as TGF-?signaling pathway and IL-17 signaling pathway.In the early infection of CD8+ T cells,the cell cycle and DNA replication pathways are significantly activated,and the proteasome and sphingolipid metabolism pathways of chronic infection are significantly activated.We used metabolic coupling to compare the changes in the metabolic pathways of the two T cell subtypes at different stages of infection.Analysis of the metabolic function of these two cells showed that the metabolic characteristics of CD8+ T cells changed more significantly than that of CD4+ T cells.And the metabolic pathways that are significantly changed after HIV infection are "degradation of valine,leucine and isoleucine","?-alanine metabolism" and "PPAR signaling pathway".These three pathways are also CD4+ T cells.The most varied pathway between early infection and chronic infection.Through gene set enrichment analysis and protein interaction prediction,we found that apoptosis-related dot-like protein(PYCARD)has an inhibitory effect on the HIV virus.Verification by molecular biology experiments,preliminary results show that PYCARD has the ability to reduce the viral load of different subtypes of HIV in both HEK293 T cells and CD4+ T cells,and PYCARD has no effect on cell viability and apoptosis.Changes in its expression content have also been found in a variety of latent cell lines(Jlat,Ach2,etc.),but it is currently tentatively believed that it will not affect the latent state.Conclusion: After HIV infection,CD4+ T cells and CD8+ T cells have different transcription characteristics.We have identified different functional modules,and identified the module features and key targets.In acute infection,the NOD-like receptor signaling pathway is a possible key signaling pathway.HIV infection destroys the metabolic balance between CD4+ T cells and CD8+ T cells.The changes in the metabolic characteristics of CD8+ T cells are more obvious than that of CD4+ T cells.PYCARD has the effect of reducing HIV viral load,and the change of latent state will change its expression.