【摘 要】
：
目的 探讨B淋巴细胞功能变化在不明原因复发性自然流产患者中的作用.方法 收集生殖中心就诊病例共48例,根据病史及生殖中心流产原因筛查结果分为A组(n=10)、B组(n=16)、C组(n=12)、D组(n=10)四组.采用淋巴细胞分离液于室温下梯度离心分离患者外周血单个核细胞(PBMC),加入特异性丝裂原(葡萄球菌蛋白A)及细胞因子(IL-2)共培养,刺激B淋巴细胞增值及分泌功能.细胞悬液培养5d后,一部分细胞悬液用MTT法检测外周血B淋巴细胞的增殖功能.于96孔板中加样200 μL,每个样品设置3个复孔,
【机 构】
：
贵州省人民医院生殖医学科,贵州贵阳550002;贵州中医药大学,贵州贵阳550002
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目的 探讨B淋巴细胞功能变化在不明原因复发性自然流产患者中的作用.方法 收集生殖中心就诊病例共48例,根据病史及生殖中心流产原因筛查结果分为A组(n=10)、B组(n=16)、C组(n=12)、D组(n=10)四组.采用淋巴细胞分离液于室温下梯度离心分离患者外周血单个核细胞(PBMC),加入特异性丝裂原(葡萄球菌蛋白A)及细胞因子(IL-2)共培养,刺激B淋巴细胞增值及分泌功能.细胞悬液培养5d后,一部分细胞悬液用MTT法检测外周血B淋巴细胞的增殖功能.于96孔板中加样200 μL,每个样品设置3个复孔,加入MTT溶液20 μL,并于培养箱中培养4h后,以调零组调零检测其在490 nm处OD值.留取另一部分培养后的细胞悬液采用E-LISA法检测外周血B淋巴细胞的分泌功能,通过加样、温育、洗涤、加酶等步骤在450 nm处检测其OD值并根据标准品浓度及OD值绘制标准曲线,根据标准曲线计算不同患者外周血中B淋巴细胞的分泌功能.结果 A、B两组患者其外周血B淋巴细胞经过葡萄球菌蛋白A及IL-2刺激后,其增殖效果较C、D两组显著提高,且差异有统计学意义(P＜0.05).但A、B两组之间(P=0.394),C、D两组之(P=0.786)其增殖差异并无统计学意义.B组较之C组,其分泌IgG的能力增强,且差异有统计学意义(P=0.030).结论 母体体液免疫调节失衡可能是不明原因复发性自然流产的一个重要因素.
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