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CD4~+CD25~+调节性T细胞诱导同种异体复合组织移植免疫耐受功能的研究

Study on the effect of CD4~+ CD25~+ regulatory T cell adoptive transfusion on humoral immune function in rat composite tissue allotransplantation model

摘要:

目的 探讨供体抗原特异性CD4~+CD25~+调节性T细胞(Treg),对同种异体复合组织 移植(CTA)受体大鼠细胞免疫耐受功能的影响.方法 采用免疫磁珠法分选CD4~+CD25~+Treg,将数 量1×10~6的CD4~+CD25~+Treg输注入CTA受体的大鼠,术后30 d采取外周血,尼龙毛柱分离T、B细 胞,检测在IgM 抗体刺激下的增殖放射强度的液体闪烁测定(CPM)值及血清lgG和IgA水平,判断 CD4~+CD25~+Treg对T、B细胞功能的影响及CTA的存活情况,并进行统计学分析.结果 分选的CD4~+CD25~+Treg纯度为95.6%.T、B细胞CPM值:正常对照组分别为2436±358、2418±348,Treg 局部干预组为2273±136、2252±127,Treg全身干预组分别为2338±228、2315±218,Treg未干预组分 别为3749±245、3720±224;Treg 未干预组与其三组比较差异均有统计学意义(P<0.01).血清IsG、IgA水平:Treg 局部和全身干预组分别为(12.56±1.30)g/L、(2.38±0.21)g/L和(13.48±1.23) g/L、(2.86±0.24)g/L,与正常对照组(12.35±1.28)g/L、(2.36±0.12)g/L比较差异无统计学意 义(P>0.05),三组与Treg未干预组(16.58±1.12)g/L、(3.75±0.37)g/L比较差异有统计学意义 (P<0.01).Treg 局部和全身干预组CTA存活时间分别为(97±13)和(63±lO)d,显著长于Treg未 干预组的(22±8)d(P<0.01).结论 供体抗原特异性CD4~+CD25~+Treg对T、B细胞功能有明显的 抑制作用,能诱导CTA免疫耐受,延长其存活时间,且局部应用效果优于全身.

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abstracts:

Objective To approach the effect of the donor antigenic specificity CD4~+ CD25~+ regulatory T cell(Treg) on cellular immune tolerance function in rat composite tissue allotransplantation (CTA).Methods Use the method of immunomagnetic beads to separate CD4~+CD25~+ Treg.(1×10°) CD4~+ CD25~+ Treg was transfused to rat CTA model.Collected pefipheral blood 30 days after operation.and used nylon WooI column to separate B cell and T cell.With the stimulation of IgM.detected B cell proliferation and the level of IgG and lgA in serum Observed the effect of CD4~+ CD25~+Treg on B cell and T cell function and the survial of allotransplants,and analyzed the data by statistics.Results The purity of separated CD4~+ CD25~+ Treg was 95.6%.The CPM of T cell of normal control group.topical intervention group,systemic intervention group and non-interventiom group were(2436±358),(2273±136),(2338± 228)and(3749±245).The CPM of B cells of normal control group,topical intervention group.systemic intervention group and non-interventiom group were(2418±348),(2252±127),(2315±218) and (3720±224),there was a significant difference in these groups (P<0.01).The serum level of IgG and lgA of topical intervention group and systemic intervention group were(12.56±1.30),(2.38±0.21), (13.48±1.23)and(2.86 4±0.24)g/L,and of normal control group was(12.35±1.28),(2.36± 0.12)g/L,had no significant difference (P>0.05).But Treg of non-intervention group was (16.58± 1.12),(3.75±0.37)g/L,there was a significant difference in the non-intervention group and the three above groups(P<0.01).The survival time of CTA in intervention of local and systemic groups were (97±13)and(63±10)d,which were significant longer than the non-intervention group[(22±8)d,P< 0.01].Conclusions Donor antigen specific CD4~+ CD25~+ Treg has significantly inhibited B cell and T cell function.It can induce immune tolerance and extend the survival time of CTA:as well local application iS better than systemic.

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