去除IgG的系统性红斑狼疮血小板减少血清对脐带血巨核细胞克隆形成单位生成的影响

目的: 研究去除 IgG 的系统性红斑狼疮(SLE) 合并血小板减少患者的血清, 对正常脐带血巨核细胞克隆形成单位(CFU-MK) 生成的影响。方法: 建立正常人脐带血单个核细胞在血小板生成素(TPO) 等作用下, 向 CFU-MK 分化的半固体培养体系;亲和层析去除 SLE 血小板减少患者血清中的 IgG 组分并加入培养体系, 研究干预前后CFU-MK集落形成的数量和形态学差异 。结果: 正常脐带血(n =10) CFU-MK 集落数为(69.3 ±19.4) 个/片, 多为中~大集落 ;加入 SLE 血小板减少患者血清(n =4) 后, 集落数为(58.5±32.5) 个/片, 与正常相比变化相近(P >0.05) , 但其中一例患者减少为 29 个/片;去除血清 IgG 组分后(n =4) ,CFU-MK 为(73.0±28.3) 个/片, 与正常相比变化相近(P >0.05) ;其中一例加入血清后减少为35 个/片, 去除 IgG 后仍为 31 个/片, CFU-MK 生成抑制未解除。形态学上, 加患者血清后大集落数由正常对照的(37.4 ±10.9) 个/片显著减少为(9.0±8.8) 个/片(P<0.05) ;去除 IgG 后, 大集落数亦显著减少为(8.3±3.9) 个/片(P<0.05) 。而加入患者血清或去除 IgG 后, 小集落相对数量比正常对照显著增多(P<0.05) 。结论: SLE 血小板减少患者血清中可能存在阻碍 CFU-MK 分化发育的抑制物, 这种抑制物可能不依赖于血清 IgG 组分。

Effects of serum without immunoglobulin G from patients with systemic lupus erythematosus associated thrombocytopenia on megakaryocyto-poiesis of normal stem cells derived from cord blood

AIM: To explore the contribution of serum without immunoglobulin G from patients with systemic lupus erythematosus (SLE) associated throm-bocytopenia on megakaryocytopoiesis of stem cells de-rived from normal cord blood .METHODS: The im-munoglobulin G was depleted from serum of patients with SLE associated thrombocytopenia by Affinity Chro-matography .The mononuclear cells derived from nor-mal cord blood were cultured in semi-solid culture sys-tem in vitro in presence of thrombopoietin (TPO) . Colony forming units-megakaryocyte (CFU-MK) was quantified by an indirect immunophosphatase alkaline labeling technique using an anti-GP Ⅱb/Ⅲa MoAb. RESULTS: No statistically difference can be found be-tween the number of CFU-MK (69.3 ±19 .4) in nor-mal control(n =10) and the group(n =4) with addit-ional serum from SLE patients with thrombocytopenia or without immunoglobulin G (58.5 ±32.5 vs 73.0 ± 28.3, P >0.05) Suppression of megakaryocyto-poiesis, however, could be observed only in one case after addition of serum, and the suppression did not disappear when immunoglobulin G was depleted from the serum by Affinity Chromatography .Morphological-ly, the large colony could be observed in normal cord blood, while almost all of colony was small after addit-ion of serum of patients (13.9±3 .9 vs 34.0±12.6, P<0.05) or without immunoglobulin G (13.9 ±3.9 vs 43.8 ±15.4, P<0.05) compare to normal con-trol .CONCLUSION: It may suggest that the serum from patients with SLE associated thrombocytopenia could suppress the formation of CFU-MK derived from normal cord blood.The suppression might be indepen-dented on immunoglobulinG .