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目的:采用硫酸铵沉淀法从中药槲寄生中提取制备分子量约为31ku的槲寄生凝集素有效成分,观察其对诱导正常人外周血γδ-T细胞受体(γδ-TCR)阳性T细胞表达、杀伤Daudi细胞和增强肿瘤组织中浸润的γδ-T细胞特异杀伤肿瘤细胞的能力。方法:饱和硫酸铵沉淀法获得槲寄生凝集素;免疫荧光法检测γδ-TCR阳性细胞表达;乳酸脱氢酶法(LDH)检测γδ-TCR杀伤靶细胞活性。结果:不同浓度槲寄生凝集素均能诱导正常人外周γδ-TCR的表达;提取的槲寄生凝集素具有诱导外周血γδ-TCR阳性细胞杀伤Daudi细胞的能力,且呈剂量依赖;槲寄生凝集素可增强肿瘤组织中γδ-TCR阳性细胞的抗瘤效应。结论:槲寄生凝集素具增强γδ-T细胞杀伤肿瘤靶细胞的生物学作用。
OBJECTIVE: To extract the effective components of the mistletoe lectin with a molecular weight of about 31ku from the Chinese mistletoe by ammonium sulfate precipitation and observe its effect on the expression of γδ-TCR positive T cells in normal human peripheral blood. The ability to kill Daudi cells and enhance the infiltration of γδ-T cells in tumor tissue specifically killing tumor cells. Methods: The mistletoe lectin was obtained by ammonium sulfate precipitation. The expression of γδ-TCR positive cells was detected by immunofluorescence. The target cell activity of γδ-TCR was detected by lactate dehydrogenase (LDH) assay. Results: Lectin at different concentrations could induce the expression of γδ-TCR in normal human peripheral blood. The extracted mistletoe lectin had the ability of inducing peripheral blood γδ-TCR positive cells to kill Daudi cells in a dose-dependent manner. Mistletoe lectin Can enhance the anti-tumor effect of γδ-TCR positive cells in tumor tissue. Conclusion: Mistletoe agglutinin has the biological effect of enhancing γδ-T cell killing tumor target cells.