Polarization Of Macrophage Induced By Chinese 1 Toxoplasma Gondii During Pregnancy And The Role Of Rhoptry Protein 16 In Cell Apoptosis

Objective:To investigate the differential bias of peritoneal macrophages during Toxoplasma gondii infection in rats pregnancies; and to define the role of rhoptry protein 16(ROP16) of Wh3, a strain of Chinese 1 type which predominantly prevalent in China in cell apoptosis.Methods:The female Sprague-Dawley(SD) rats were randomly divided into 4 groups : control group without pregnancy(con-1), control group with pregnancy(con-2), infection group of pre-pregnancy(inf-1), infection group in pregnancy(inf-2). The two infection groups were injected intraperitoneally with 2000 tachyzoites of Toxoplasma Wh6 strain in 1ml of 0.9% sterile saline solution before the day when females were housed with males, and on gestation day 8(GD8), respectively. The two groups of control were given 1ml 0.9% sterile saline solution only. On GD14, the rats were killed under euthanasia and peritoneal macrophages were harveste. Production of M1 and M2 associated cytokines were measured by ELISA and western blottin. The m RNA expression of M1 and M2 cytkones were measured by q RT-PCR. The recombinant eukaryotic expression vector p EGFP-ROP16 was transfected into 293 T cells. Apoptosisi-associated gene PCR arrays were carried out in the transfected eukaryote. Western blotting and flow cytometry were performed to observe the potentially inhibitory apoptosis resistance in p EGFP-ROP16 transfected cells.Results:Inf-1 group had the highest levels of NO, IL-10 and TNF-α. However, Arg-1, IL-12 and TGF-β levels were higher in inf-2 group. Notably, NO was significantly reduced in group con-2. The recombinant eukaryotic expression vector p EGFP-ROP16 was transfected into 293 T cells. Apoptosis associated gene PCR array showed that 11 genes(LTA, TNFSF11 B, BAD et al.) were up-regulated by ROP16 transfection. Western blotting revealed Caspase-3, Caspase-9, and PARP-1 do not make significant difference in the three groups.Conclusion:The peritoneal macrophage of normal pregnancy rats was polarized toward M2, and the peritoneal macrophage infection wirh T.gondii pre-pregnancy was polarized toward M1, while infection wirh T.gondii in-preganacy induced the peritoneal macrophage polarize to M1 and activated M2 at the same time. These results suggested that infection with T.gondii pre-pregnancy might cause a worse pregnancy outcome, as it interfered the immune tolerance of pregnancy.ROP16 of Wh3 strain regulated 293 T cell apoptosis only at the molecular level lightly, but no obvious change at the cellular level.