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目的研究人骨髓间充质干细胞(MSC)体外对 B 淋巴细胞的免疫调节作用。方法从人骨髓中分离培养 MSC,通过观察细胞形态和流式细胞术检测细胞表面标志进行鉴定。用加速器辐射去除 MSC 增殖分化能力。检测加入骨髓 MSC 前后 B 淋巴细胞增殖能力变化,并对其进行凋亡分析;比较 Transwell 非接触培养与直接接触培养中,MSC 对活化 B 淋巴细胞增殖的作用;观察骨髓 MSC作用前后,活化 B 淋巴细胞分泌免疫球蛋白 IgG、IgA 和 IgM 的能力,以及 B 淋巴细胞表面免疫分子的表达。结果①MSC 不能刺激 B 淋巴细胞增殖;MSC 抑制 LPS 活化的 B 淋巴细胞增殖,其抑制作用与浓度呈依赖性。②MSC 不能诱导 B 淋巴细胞凋亡;Transwell 非直接接触培养组中 MSC 抑制 B 淋巴细胞增殖的作用较直接接触培养组的作用弱,提示 MSC 抑制 B 淋巴细胞增殖的作用包括物理性接触抑制和非接触抑制。③骨髓 MSC 抑制活化的 B 淋巴细胞分泌免疫球蛋白 IgG、IgM 和 IgA;MSC 作用下,B 淋巴细胞表面 HLA-DR、CD40、CD80和 CD86的表达无显著改变。结论骨髓 MSC 在体外可抑制 B淋巴细胞增殖和分泌功能,具有一定的免疫调节能力。
Objective To study the immunomodulatory effects of human bone marrow mesenchymal stem cells (MSCs) on B lymphocytes in vitro. Methods MSCs were isolated and cultured from human bone marrow. Cell morphology was observed by flow cytometry and cell surface markers were identified by flow cytometry. Accelerated irradiation removes MSC proliferation and differentiation. The proliferation of B lymphocytes was detected before and after the addition of bone marrow MSCs, and the apoptosis of B lymphocytes was analyzed. The effect of MSC on the proliferation of activated B lymphocytes was compared between Transwell non-contact culture and direct contact culture. The ability of cells to secrete immunoglobulin IgG, IgA, and IgM, as well as the expression of B lymphocyte surface immune molecules. Results ① MSC could not stimulate the proliferation of B lymphocytes; MSC inhibited the proliferation of B lymphocytes activated by LPS, and its inhibitory effect was dependent on the concentration. MSCs could not induce apoptosis of B lymphocytes; the effect of MSC inhibiting B lymphocyte proliferation in Transwell non-direct contact culture group was weaker than direct contact culture group, suggesting that the effect of MSC in inhibiting B lymphocyte proliferation includes physical contact inhibition and non-contact inhibition. ③ BMSCs inhibited the secretion of immunoglobulin IgG, IgM and IgA by activated B lymphocytes. There was no significant change in the expression of HLA-DR, CD40, CD80 and CD86 on B lymphocytes under the action of MSC. Conclusion Bone marrow MSCs can inhibit the proliferation and secretion of B lymphocytes in vitro and have certain immunomodulatory capacity.