巨核细胞的免疫学研究

The Immunological Study of Megakaryocytes

巨核细胞的免疫学作用鲜被研究 ,本研究观察巨核细胞的粘附分子和细胞因子表达 ,以及细胞因子与巨核细胞的相互作用 ,以了解巨核细胞的免疫学作用。使用流式细胞术 ,ELISA和免疫组化方法 ,分析粘附分子CD36在血小板、骨髓巨核细胞和巨核细胞株 (Meg 0 1,Dami,CHRF 2 88 11和M 0 7e)的表达 ;使用RT PCR方法 ,检测白细胞介素 1(IL 1)至白细胞介素 10 (IL 10 ) ,TNF α ,TNF γ和IFN γ在 4个巨核细胞株的表达 ;使用血浆凝块巨核细胞集落培养方法 ,观察细胞因子IL 1β ,IL 3,IL 6和TPO对小鼠巨核细胞生长的影响 ,乙酰胆碱酯酶染色鉴定和识别巨核细胞。结果 :①血小板、骨髓巨核细胞和巨核细胞株均表达CD36 ,而且随着细胞的发育成熟 ,CD36表达越来越高 ,提示巨核细胞通过合成血小板膜上的粘附蛋白参与免疫反应 ;② 4个巨核细胞株代表不同成熟阶段巨核细胞的模型 ,炎性细胞因子TNF α ,IL 1β ,IL 3和IL 6在 4个细胞株均有表达 ,提示不同阶段的巨核细胞均可作为炎性细胞因子的一个来源 ,参与机体的免疫过程 ;③细胞因子IL 1β ,IL 3和IL 6对巨核细胞生长均有不同程度的促进活性 ,尽管较血小板生成素低 ,仍提示巨核细胞存在一个自分泌 (autocrine)反应 ,影响自身的生长。

The immunological role of megakaryocytes is not well known. This project studies the involvement of megakaryocytes on immuno-inflammatory processes and the possible mechanism via the adhesion molecule CD36 and the synthesis of relevant cytokines. The expression of adhesion protein CD36 on human platelets, megakaryocytes and megakaryocytic cell lines (Meg-01, Dami, CHRF-288-11 and M-07e) was analyzed by using flow cytometry, ELISA and immunocytochemical methods. The expression of interleukin-1 (IL-1) to interleukin-10 (IL-10), TNF-α,TNF-γ and IFN-γ on the four megakaryocytic cell lines was also determined by RT-PCR. The effect of IL-1β, IL-3, IL-6 and TPO on murine megakaryocyte colony formation (CFU-MK) was studied by using a plasma clot culture system. The CFU-MK was confirmed by acetylcholine esterase staining. The results showed that: ① CD36 was expressed on platelets, megakaryocytes and the four megakaryocytic cell lines, the relative expression level is as follows: platelets > megakaryocytes > Meg-01 >Dami > CHRF-288-11 > M-07e, suggesting that the level of CD36 expression correlates with the degree of maturity of megakaryocytic differentiation; ② inflammatory cytokines TNF-α, IL-1β, IL-3 and IL-6 were detected in all the four megakaryocytic cell lines, suggesting that different stages of megakaryocytes can be as a source of inflammatory cytokines; and ③ IL-1β, IL-3 and IL-6, as well as TPO, play a stimulating effect on CFU-MK formation, suggesting that there is an 'autocrine' effect on megakaryocytopoiesis. The data obtained suggest that megakaryocytes may involve in immuno-inflammatory processes via the synthesis of platelet adhesion molecules and inflammatory cytokines.