海胆黄多糖SEP对小鼠T、B淋巴细胞的选择性激活作用.docVIP

  海胆黄多糖 SEP 对小鼠 T、B 淋巴细胞的选 择性激活作用# 5 10 15 20 25 30 35 40  摘要：目的：研究海胆黄多糖 SEP 对小鼠 T、B 淋巴细胞的选择性激活作用，并初步探讨其 机制。方法：尼龙毛柱法分离小鼠脾 T、B 淋巴细胞，MTT 法测定 SEP 对两种细胞增殖的 影响；ELISA 测定对 T、B 细胞因子产量的影响；RT-PCR 测定 SEP 对 T 细胞因子基因表达 的影响，流式细胞技术测定对 T 细胞亚群的影响；MTT 法测定 CD3 单抗对 SEP 刺激 T 细 胞增殖的影响。结果：SEP 对 T 细胞增殖、细胞因子产量的促进作用明显强于对 B 细胞； SEP 可以使脾细胞悬液 IL-2、IFN-γ 和 TNF-α 的基因表达显著上调，使 CD3+细胞和 CD4+/CD3+的百分比显著增加；CD3 单抗能抑制 SEP 对 T 淋巴细胞的增殖作用。结论：SEP 主要选择性地直接激活小鼠 T 细胞，对 B 细胞的影响则较小；SEP 可以直接通过 Th1 反应 激活 T 淋巴细胞；CD3 是 SEP 激活小鼠 T 淋巴细胞的一个可能的受体。 关键词：生化药学；海胆黄多糖 SEP；T 淋巴细胞；B 淋巴细胞；激活作用；机制 中图分类号：R9 Selective activation of mouse T and B lymphocytes by polysaccharide SEP from Strongylocentrotus nudus eggs LIU Chunhui, CAO Jichao, CHEN Li (School of Pharmaceutical Sciences National Glycoengineering Research Center, Shandong University, Jinan 250012) Abstract: Objective To investigate the selective activation mouse T and B lymphocytes by polysaccharide SEP and its mechanism. Methods The effects of SEP on mouse T cell and B cell proliferation were determined by MTT assay after the cells were purified on nylon wool column, the productions of T- and B- lymphocytic cytokine were tested using ELISA kits, the effects of SEP on cytokine mRNA expression in splenocytes were determined by RT-PCR, the effects on T lymphocyte subsets were analyzed by flow cytometry, and the effects of CD3 monoclonal antibodies on SEP-inducing T lymphocyte proliferation were also determined by MTT assay. Results SEP stimulated both T cell and B cell proliferation and cytokine production. However, the influence on B cells was greatly weaker than that on T cells. SEP up-regulated mRNA expression for T cell cytokine and increased the percentages of CD3+ and CD4+/CD3+ lymphocyte subsets. CD3 monoclonal antibodies inhibited SEP-stimulating T lymphocyte proliferation. Conclusion SEP mainly activated T lymphocyte by Th1 response, which was related to cell surface receptor, such as CD3. Key words: biochemical pharmacy; Strongylocentrotus nudus egg polysaccha