The Functionary Mechanism Of A20 And ABIN1 In The NF-?B Signaling Pathway On The Proliferation And Apoptosis Of Human Leukemia T Cells Induced By Toxoplasma Gondii

ObjectiveTaking Toxoplasma gondii ME-49 infection to human T cell line Jurkat T cells and Molt-4 T cells as a model,and taking NF-?B pathway and its related regulatory proteins as the research object,to analyze the effect of A20 on the proliferation and apoptosis of human T cells induced by Toxoplasma gondii and the role of ABIN1 protein in the regulation of Caspase8 protein.Methods1.Tunel assay,Flow cytometry assay were used to detect the apoptotic effect of human leukaemia T-cell lines Jurkat and Molt-4.2.Western blotting was used to detect the protein level of p NF-?B,A20,ABIN1 after T.gondii ME-49 infection.3.Lentiviruses sh RNA were used to knockdown the A20 and ABIN1 gene,and then detect the expression level of each protein by using western blotting after knockdown.4.The overexpression-plasmid target ABIN1 gene was transfected into Jurkat T cell and Molt-4 T cell,and then detect the expression level of each protein by using western blotting.Rusults1.Under the infection of a certain number of Toxoplasma gondii ME-49 tachygonite,the proliferation effect of Jurkat T cell and Molt-4 T cell were inhibited,and the level of apoptosis increased significantly,and it increased significantly with the increase of tachychon number(5×105,1×106,5×106).2.Under the infection of Toxoplasma gondii ME-49,the expression level of NF-?B and ABIN1 protein Jurkat in T cells was significantly down-regulated with the increase of tachyzoites number(5×105,1×106,5×106),and the expression level of A20 was significantly increased(P<0.05)with the increase of tachyzoites number(5×105,1×106,5×106),the similar changes were present in Molt-4 T cell.3.Using lentiviral sh RNA to knockdown the A20 gene of Jurkat T cells,the protein level of A20 was significantly down-regulated.And under the infection of Toxoplasma gondii ME-49(5 ×106),the NF-kappa B and ABIN1 protein levels were up-regulated in Jurkat T cell(P<0.05),the protein level of Caspase-8 was significantly down-regulated(P<0.05),and the level of apoptosis decreased,the similar changes were present in Molt-4 T cell.4.Using lentiviral sh RNA to knockdown the ABIN1 gene of Jurkat T cells,the expression level of ABIN1 was significantly decreased(P<0.05).And under the infection of Toxoplasma gondii ME-49(5×106),the level of Caspase-8 was significantly increased(P<0.05),and the level of apoptosis also increased significantly,the similar changes were present in Molt-4 T cell.5.The overexpression-plasmid target ABIN1 gene was transfected into Jurkat T cell,the expression level of ABIN1 was significantly increased(P<0.05).And under the infection of Toxoplasma gondii ME-49(5×106),the level of Caspase-8 was significantly increased(P<0.05),and the level of apoptosis also decreased significantly,the similar changes were present in Molt-4 T cell.Conclusion1.Toxoplasma gondii ME-49 infection inhibited the proliferation and induced apoptosis through down-regulate the expression level of phosphorylation NF-?B in T cell lines.2.Toxoplasma gondii ME-49 could up-regulate the level of A20 protein to inhibit the activation of NF-?B pathway and the expression of ABIN1.3.Increasing ABIN1 protein could down-regulate the expression of caspase-8 protein and resist the apoptosis of human leukemia T cells induced by Toxoplasma gondii.