Research On The Disorders Of The NKT Cells In The Unexplained Recurrent Spontaneous Abortions Mouse Model

ObjectiveComparative Study of the NKT cells numbers,cytokine secretion,maturity and expression levels of T-bet in mice trophoblast with normal pregnancy and unexplained recurrent spontaneous abortion.MethodsEstablish normal pregnancy model by feeding CBA / J and BABL/C in a cage, unexplained recurrent spontaneous abortions model by feeding CBA / J and DBA/2J in a cage. Observe the vagina embolization in mice daily. In the 9,11,13,15 days,terminate pregnancy. Grind the trophoblast wire on the mesh 200 to get cells suspension. Crack erythrocytes. Stain 1.5×106 cells with the CD3,α-loaded CD1d Galcer tetramer, CD69 antibody, then detected by flow cytometry. Culture 5×106 cells in RPMI1640 +15% FBS medium, two days after the collection medium supernatant, and the suspension cells and from the total RNA, reverse transcriptase into cDNA, fluorescence quantitative PCR analysis, testing T - bet in relation to the expression of HPRT.Results1. Normal pregnancy and unexplained recurrent spontaneous abortion group, in the course of pregnancy, classical CD3 + T cells without significant changes. Moreover, there is no significant difference between the two groups.2.Normal pregnancy group, in pregnancy process, the trophoblast cells secreting IFN-γdecreased gradually;the number and mature frequency gradually declining.3.Unexplained recurrent spontaneous abortion group, in pregnancy process, trophoblastic cells secreting IFN-γincreased gradually, the number and mature frequency of NKT cells also increased gradually.4. No matter the normal pregnancy group or the unexplained recurrent spontaneous abortion group, trophoblast cells IFN-γsecretion is related to NKT cells mature frequency.5.There is a difference of T-bet mRNA expression between Unexplained recurrent spontaneous abortion group compared with normal pregnancy,. T-bet mRNA expression levels is related to the frequency of mature NKT cells and trophoblastic cells IFN-γsecretion.Conclusions1. In the course of pregnancy, NKT cells may be the major secreting cells in the trophoblast.2.Unexplained recurrent spontaneous abortion occurred, may be related to NKT cells disorders. NKT cells have low- mature frequency and inadequate secretion of IFN-γduring early pregnancy; whereas have high- mature frequency and excessive secretion of IFN-γduring latter pregnancy.3.The anomaly of T-bet mRNA expression may be one of the factors lead to NKT cells disorder.