| Objective: Helper T cells are found in two distinct cell types, Th1 and Th2 distinguished by the cytokines they produce and respond to and the immune responses they are involved in. Th1 cells produce pro-inflammatory cytokines like IFN-γ, TNF-βand IL-2, while Th2 cells produce the cytokines like IL-4, IL-5, IL-10,IL-6 and IL-13. Th1- and Th2-helper cells direct different immune response pathways. Th1 cells drive the type-1 pathway ("cellular immunity" ) to fight viruses and other intracellular pathogens, eliminate cancerous cells, stimulate delayed-type hypersensitivity (DTH) reactions and occur organ specific autoimmune diseases. Th2 cells drive the type-2 pathway ( " humoral immunity " ) to produce humoral immune response, irritability response and extracellular infectious diseases, autoimmune diseases and be credited with tolerance of xenografts.In immunity of hematopoietic stem cell transplantation, recent results have suggested that acute GVHD is mediated by TH1 lymphocytes and that TH2 lymphocytes may exert a protective effect but may have a role in mediating chronic GHVD. The study of the regulation mechanism to T lymphocyte differentiation can help us elucidating the pathogenesis of GVHD immune response.T-BET is important transcription factor to T lymphocytes differentiation, it can make T lymphocytes emerge TH1 differentiation, while GATA3 is also important transcription factor to T lymphocytes differentiation, it can make T lymphocytes emerge TH2 differentiation. The Cells pathways MAPK and AKT/PI3K which influence T cell differentiation is also the important Cells pathways of PTEN(phosphatase and tensinhomolog deleted on chromosome ten).Jurkat cells is human T lymphoma cell line,it could secrete IFN-γand IL-4,therefore it could become the researching object for T cell differentiation .Through transfecting recombinant adenovirus vector-PTEN (Ad-PTEN) gene containing green fluorescent protein (Ad-PTEN-GFP) or the empty adenovirus vector containing green fluorescent protein (Ad-GFP) into jurakt cells ,to detect the influence on jurkat cell by PTEN gene.Method:The recombinant adenovirus containing green fluorescent protein (GFP) gene and PTEN gene (Ad-PTEN-GFP) or empty vector (Ad-GFP) was transfected into jurkat cells. Then the PTEN T-BET GATA-3 IL4 and IFN- gamma mRNA expression levels were measured by RT-PCR.Results:1 When jurkat cells were transfected with Ad-PTEN-GFP or Ad-GFP for 48 hours at MOI=200, the PTEN mRNA expression level in Ad-PTEN-GFP group (0.702±0.1) was higher than that in the non-treatment group (0.307±0.03) and in the Ad-GFP transfected group (0.306±0.0.02), (P<0.05). While there was no significant difference between in the non-treatment group and in the Ad-GFP transfected group (P> 0.05).2 When jurkat cells were transfected with Ad-PTEN-GFP or Ad-GFP for 48 hours at MOI=200, the T-bet mRNA expression level in Ad-PTEN-GFP group (0.603±0.1) was higher than that in the non-treatment group (0.323±0.02) and in the Ad-GFP transfected group (0.319±0.04), (P<0.05). While there was no significant difference between in the non-treatment group and in the Ad-GFP transfected group (P> 0.05).3 When jurkat cells were transfected with Ad-PTEN-GFP or Ad-GFP for 48 hours at MOI=200, the IFN-gama mRNA expression level in Ad-PTEN-GFP group (0.613±0.01) was higher than that in the non-treatment group (0.345±0.02) and in the Ad-GFP transfected group (0.343±0.04), (P<0.05). While there was no significant difference between in the non-treatment group and in the Ad-GFP transfected group (P> 0.05).4 When jurkat cells were transfected with Ad-PTEN-GFP or Ad-GFP for 48 hours at MOI=200, the GATA3 mRNA expression level in Ad-PTEN-GFP group (0.132±0.02) was higher than that in the non-treatment group (0.396±0.05) and in the Ad-GFP transfected group ((0.389±0.06), (P<0.05). While there was no significant difference between tnat in the non-treatment group and in the Ad-GFP transfected group (P> 0.05).5 When jurkat cells were transfected with Ad-PTEN-GFP or Ad-GFP for 48 hours at MOI=200, the IL4 mRNA expression level in Ad-PTEN-GFP group (0.289±0.01) was higher than that in the non-treatment group (0.851±0.02) and in the Ad-GFP transfected group (0.848±0.01), (P<0.05). While there was no significant difference between in the non-treatment group and in the Ad-GFP transfected group (P> 0.05).Conclusion: The recombinant adenovirus containing green fluorescent protein (GFP) gene and PTEN gene (Ad-PTEN-GFP) or empty vector (Ad-GFP) was successful transfected into jurkat cells,and the expression of the PTEN gene in Ad-PTEN-GFP group was much higher than in the control group.The high expression of PTEN gene in jurkat cells make the expression of T-bet and IFN-gama significantly elevate ,while the expression of GATA3 and IL4 descend. These results reveal that PTEN gene can make T cells differentiate toTH1 direction...
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