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Investigating the CTLA-4Ig Inducing Th17/Treg Imbalance in Peripheral Blood with Unexplained Recurrent Spontaneous Abortion Patients

Author: ZhangYingXin
Tutor: LiuYuSheng
School: Anhui Medical University,
Course: Obstetrics and Gynaecology
Keywords: CTLA-4Ig Th17 Treg URSA
CLC: R714.21
Type: Master's thesis
Year: 2011
Downloads: 57
Quote: 0
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Abstract


Objective: To observe the CTLA-4 fusion protein induced reasons unknown in the peripheral blood of patients with recurrent spontaneous abortion Th17/Treg cell conversion. Methods: (1) collect outpatient URSA peripheral blood of patients, extracted peripheral blood mononuclear cells (peripheral blood mononuclear cell PBMC) were divided into three groups, with RPMI 1640 medium at the same time added to a final concentration of 10ug/ml phytohemagglutinin (PHA ) dubbed PBMC cell concentration of 2 * 10 6cells/ml (200ul / well) in 96-well plates. The experimental groups were added CTLA4Ig0.4ul (the final concentration 1ug/ml) and after CTLA4Ig4ul (final concentrations 10ug/ml) and blank control group, 37 ° C in 5% CO2 incubator for 72 hours to collect 100μlPBMC CD4 IL-17A, CD4 CD25 FoxP3 cells change, to explore the role of CTLA-4Ig in the induction of reasons unknown recurrent the abortion patients Th17/Treg cells conversion analysis by flow analysis. (2) collection of out-patient peripheral blood of patients with URSA, extraction of peripheral blood mononuclear cells, adding CTLA-4 fusion protein stimulation for 72 hours, and blank control group. Extraction of total mRNA by fluorescence quantitative PCR, detect which changes Foxp3mRNA and RORγtmRNA. Investigate whether CTLA-4Ig can adjust the Treg cell-specific transcription factor FoxP3 and Th17 cell-specific transcription factor RORγt expression in peripheral blood in unexplained recurrent spontaneous abortion. Results: (1) CTLA-4Ig URSA peripheral blood of patients Treg cells can induce raised, Th17 cells decreased URSA patients with peripheral blood, pure PBMC cultured in the control group CD4 IL-17A / CD4 cell percentage (2.72 ± 0.07%) was significantly higher than CTLA-4Iglug/ml experimental group (1.16 ± 0.39%), the difference was significant (P lt; 0.05) CD4 CD25 Foxp3 / CD4 cells in the peripheral blood of patients with URSA ratio (2.30 ± 0.36%) was significantly lower than CTLA-4Iglug/ml experimental group (3.96 ± 0.52%) was significantly different (P lt; 0.05) The join CTLA-4Igl0ug/ml group with CTLA-4Ig1ug/ml group compared to the proportion of CD4 IL-17A / CD4 cells no significant difference, CTLA-4Igl0ug/ml CD4 CD25 FoxP3 / CD4 cell percentage slightly lower than the CTLA- group 4Ig1ug/ml, but no significant difference (P gt; O.05). In the maintenance of pregnancy speculated Th17/Treg cells may be in a relatively balanced state, once the imbalance is not conducive to pregnancy, may be one of the reasons that cause recurrent miscarriage. (2) quantitative PCR amplification of FoxP3 mRNA and RORγt mRNA changes, join the CTLA-4Ig1ug/ml experimental group RORγt mRNA 2-ΔΔCt value (0.306 ± 0.079), the expression of RORγtmRNA lower than the control group, the difference was significant (P lt ; 0.05). Same CTLA-4Ig group FoxP3 mRNA2-ΔΔCt value (5.162 ± 3.595), FOXP3 mRNA expression compared to the control group was significantly higher, the difference was significant (P lt; 0.05) Speculated that CTLA-4Ig may affect Th17 cell-specific transcription factor RORγt reduced expression of Treg-specific transcription factor FoxP3 expression was elevated from the molecular level. Conclusion: Reproductive immunology view the normal embryo is not rejection by the maternal immune tolerance depends on the maternal-fetal interface, will result in the occurrence of URSA Once this pattern is broken. Peripheral blood of patients with normal pregnancy increased number of Treg cells is conducive to the maintenance of pregnancy, and Treg reduction in the number of cells in the peripheral blood of patients with URSA, accompanied by an increase in Th17 cells in peripheral blood, is detrimental for the maintenance of pregnancy, resulting in the occurrence of abortion. So we can see that in addition to the balance between Th1/Th2 ratio between the Th17/Treg cell imbalance may also lead to the URSA occurrence. CTLA-4 plays an important role in T cell activation and differentiation of Th cell subsets and maternal-fetal immune tolerance induction. The URSA can closed CTLA-4 in peripheral blood of patients with Th2 cytokines to a Thl-type cytokines offset is not conducive to the maintenance of pregnancy Exogenous administration of a substance with the same effect of CTLA-4, such as CTLA4Ig, and may increase the proliferation of Treg cells, inhibit the development and differentiation of Th17 cells, thus contributing to the maintenance of pregnancy. Our study found that CTLA-4Ig can be induced in vitro the conversion between the URSA peripheral blood of patients Th17/Treg cells, and affect the specific transcription factor expression at the molecular level, which indicated that CTLA-4Ig URSA patients may be reversed the the imbalance between of the Th17/Treg cell, make it to offset the Treg cells are formed in the body conducive to pregnancy to maintain the environment and thus contribute to relapse treatment of abortion, provide a favorable theoretical basis for the clinical application of CTLA4-Ig.

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CLC: > Medicine, health > Obstetrics and Gynaecology > Obstetrics > Pathological pregnancy ( abnormal pregnancy ) > Miscarriage, premature labor and prolonged pregnancy
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