| ObjectiveTo explore effect of CTLA-4Ig on the periphera Th17/Treg cells and pregnantoutcomes in abortion prone matings.Methods①The female CBA/J×male DBA/2matings with very high fetal resorption rates wereused as the abortion prone model,while female CBA/J×male BALB/c matings wereused as the normal pregancy model with low resorption rates.After overnightcohabitation with DBA/2or BALB/c males,CBA/J females with vaginal plugs wereseprated from the malesand the day of vaginal plug detection was considered to be day0of pregnancy.②On the day4of pregnancy (implantation window),DBA/2-mated females werethen randomized and divided into two groups:one group receivied10ug/2d(0.5mg/Kg)intraperitoneally(i.p)., three times. The other group and the normal pregancy groupreceived the same amount of NS(i.p).③On the day14of gestation,the females were sacrificed,the abortion (resorption)sites were identified by their small size accompanied by their necrotic hemorrhagicappearance compared with normal embryos.The percentage of resorption was calculatedby the formula:R%=Re/(Re+F)×100,were R represents the percentage of resorptionsrelative to the total number of effective implantation sites,Re represents the number ofthe resorbed embryos,and F represents the number of viable embryos.The speen was collected for the following experiments.④Pick up lymphocytes of CBA/J females in three groups.⑤Flow cytometric analysis the change of CD4+CD25+Foxp3+and CD4+IL-17A+cell,to investigate possible role of CTLA-4Ig inducing Th17/Treg imbalance inabortion-Prone matings.⑥By RT-PCR methods for the expression of Foxp3mRNA and RORγtmRNA ofsamples.Explore whether CTLA-4Ig adjust the Treg cells specific transcription factersFoxp3and Th17cells specific transcription factor RORγt in abortion-Prone matings.Rsults①CBA/Jfemales that were mated with DBA/2males (abortion group) experiencedembryo resorption rates of25%.This was significantly higher compared with the CBA/Jfemales that were mated with BALB/2males(control group)(P<0.05).Abortion-pronefemale mice treated with CTLA-4Ig(CTLA-4Ig group) showed a significantly lowerabortion rate compared with abortion group. An embryo resorption rate of CBA/Jfemales mated with BABL/c males of about10.8%, this did not show a significantdifference compared with CTLA-4Ig-treated abortion-prone mice(P>0.05).②The proption of CD4+CD25+Foxp3+cell in abortion group showed a significantlylower than the normal group(P<0.05).However, the proption of CD4+IL-17A+cell inabortion-prone was significantly higher compared with the normal group(P<0.05).The proption of CD4+CD25+Foxp3+cell in abortion-prone CBA/J females receivingCTLA-4Ig(CTLA-4Ig group) showed higher than those from the abortion-prone CBA/Jfemales(P<0.05). The proption of CD4+IL-17A+cell in abortion-prone CBA/J femalesreceiving CTLA-4Ig(CTLA-4Ig group) showed lower than those from theabortion-prone CBA/J females(P<0.05).③Foxp3mRNA expression was significantly increased in abortion-prone CBA/Jfemales receiving CTLA-4Ig(CTLA-4Ig group)compared with the the abortion-prone CBA/J females(P<0.05). RORγtmRNA expression was significantly decreased inabortion-prone CBA/J females receiving CTLA-4Ig(CTLA-4Ig group)compared withthe the abortion-prone CBA/J females(P<0.05).CoculsionSuccessful pregnancy requires a state of immunological tolerance since normallythe maternal immune system does not reject the semi-allogeneic conceptus. Our resultsshow that CTLA-4Ig can induce the balance of Th17/Treg Cell in mice abortion modeland increase Foxp3mRNA expression，which reduces the embryonic absorption rate,and eventually improves the pregnancy outcome. |
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