胆固醇自稳失调对SP2/0骨髓瘤细胞免疫原性的影响

Effects of disturbed cholesterol homeostasis on the immunogenicity of SP2/0 myeloma cells

目的 探讨胆固醇自稳失调对SP2 0骨髓瘤细胞免疫原性的影响。方法 以去脂血清 (LDS)和 或洛伐他汀 (LOV)处理SP2 0细胞 ,导致胆固醇自稳失调 ,以密度梯度离心法分离细胞膜 ;以rmGM CSF和rmIL 4诱导获得小鼠骨髓细胞来源的DC ,混合淋巴细胞反应 (MLR)检测其功能 ;DC负载细胞膜抗原活化T细胞获得肿瘤特异性CTL ,MTT法检测其对SP2 0细胞的杀伤效果。结果 MLR结果表明DC具有较强的刺激同种T细胞增殖的能力 ;与对照组膜抗原诱导的CTL相比 ,经胆固醇自稳失调处理的SP2 0细胞膜负载DC诱导的CTL的杀伤活性增强 ,尤以高效靶比 (2 0∶1)时细胞杀伤作用增强更为明显 ,CTL细胞毒性具有肿瘤特异性。结论 胆固醇自稳失调的SP2 0细胞膜负载DC致敏的CTL的细胞毒作用增强 ,并与胆固醇失调严重程度有一致性。

Objective To explore the effects of disturbed cholesterol homeostasis on the immunogenicity of SP2/0 myeloma cells. Methods The membrane of SP2/0 cells treated with LDL-deficient serum (LDS) and/or lovastatin (LOV) was obtained by density gradient centrifugation. Bone marrow-derived dendritic cells (DCs) were generated in vitro in the presence of recombinant murine granulocyte-macro- phage colony-stimulating factor (rmGM-CSF) and recombinant murine interleukin-4 (rmIL-4), then the immunological function of the DCs were detected by mixed lymphocyte reaction (MLR). Cytotoxic T lymphocytes (CTLs) were induced by membrane antigen-loaded DCs. The cytoto- xicity of CTLs against SP2/0 cells was evaluated by MTT. Results DCs generated from murine bone marrow precursors had a potent capacity for stimulating allogenic T lymphocytes proliferation. CTLs, induced by cellular membrane antigen of LDS- and/or lovastatin-treated SP2/0 cells, have stronger tumor-specific cytotoxicity than did the CTLs induced by untreated SP2/0 cells, especially at the high effector/target ratio. Conclusion Perturbation of cholesterol homeostasis resulted from treatment of LDS and /or LOV can strengthen the cytotoxicity of CTLs induced by SP2/0 cellular membrane-loaded DCs. The intensity of cytotoxicity of CTLs was positively correlated with the levels of cell cholesterol disturbance.