灵芝多糖对体外树突状细胞诱导细胞毒性T-淋巴细胞的调节作用(英文)

目的:研究灵芝多糖(Gl-PS)在抗原提呈阶段对体外培养树突状细胞(DC)诱导特异性细胞毒性T-淋巴细胞(CTL)功能的调节及其机制。方法:体外培养小鼠骨髓来源DC经P815肿瘤细胞冻融抗原冲击致敏,并与不同浓度Gl-PS(0.8,3.2,或12.8 mg/L)共培养。成熟DC与脾淋巴细胞共培养诱导P815特异性CTL生成。第5天收集各组悬浮细胞及培养上清,采用乳酸脱氢酶法比较CTL特异性杀伤活性;RT-PCR法测定T扰素γ(IFNγ)及颗粒酶B mRNA在CTL的表达;ELISA或Western blot法检测IFNγ或颗粒酶B蛋白的表达。结果:3种浓度的Gl-PS均可增高培养上清中释放的LDH活性(P<0.01);增加CTL表达IFN7及颗粒酶B mRNA(IFNγ:Gl-PS 12.8 mg/L组与RPMI-1640组,P<0.05;颗粒酶B:P<0.01);促进CTL培养上清中IFNγ蛋白生成(P<0.05)以及CTL表达颗粒酶B蛋白(Gl-PS 12.8 mg/L组与RPMI-1640组,P<0.05)。结论:Gl-PS可在抗原提呈阶段促进P815肿瘤冻融抗原冲击致敏DC所诱导的特异性CTL的杀伤活性,其机制可能是通过IFNγ及颗粒酶B途径的调节。

Regulatory effect of Ganoderma lucidum polysaccharides on cytotoxic T-lymphocytes induced by dendritic cells in vitro

AIM: To study the regulatory effects of Ganoderma lucidum polysaccharides (Gl-PS) on cytotoxicity and mechanism of specific cytotoxic T-lymphocytes (CTL) induced by dendritic cells (DC) in vitro during the stage of antigen presentation. METHODS: Cultured murine bone marrow-derived DC were pulsed with P815 tumor cell lysates and co-incubated with or without various concentrations of Gl-PS (0.8, 3.2, or 12.8 mg/L) at the same time. P815 specific CTL were induced by spleen lymphocytes stimulated with mature DC. Non-adherent cells and culture supernatants were harvested on d 5 for analysis of specific cytotoxicity with lactate dehydrogenase (LDH) activity assay, mRNA expression of IFNgamma, granzyme B with RT-PCR assay, and protein expression of IFNgamma, granzyme B with ELISA or Western blot assay, respectively. RESULTS: Three concentrations of Gl-PS promoted LDH activities released into culture supernatants (P<0.01). It also increased mRNA expression of IFNgamma in CTL (Gl-PS 12.8 mg/L vs RPMI medium 1640, P<0.05) and granzyme B in CTL (P<0.01). Protein production of IFNgamma in culture supernatants (P<0.05) and protein expression of granzyme B in CTL (Gl-PS 12.8 mg/L vs RPMI medium 1640, P<0.05) were also augmented by Gl-PS. CONCLUSION: Gl-PS is shown to promote the cytotoxicity of specific CTL induced by DC which were pulsed with P815 tumor antigen during the stage of antigen presentation, and the mechanism of cytotoxicity is believed to be going through IFNgamma and granzyme B pathways.