目的 探讨抗内皮细胞抗体(AECA)介导的血管内皮(EC)损伤在异基因造血干细胞移植(allo-HSCT)中的病理生理机制.方法 收集allo-HSCT后患者的血清并纯化IgG型免疫球蛋白,将后者与人脐静脉内皮细胞(HUVEC)共培养,观察HUVEC的功能变化及凋亡情况.结果 HUVEC与AECA阳性的IgG共培养后,黏附分子表达显著增高:IgG浓度为160、320、640 μg/ml时,上清可溶性胞间黏附分子-1浓度分别为(117.10±12.82)对(78.17±4.90)pg/ml,(151.30±15.35)对(89.46±6.02)pg/ml,(239.00±32.53)对(127.8±13.86) pg/ml(P值均<0.01);IgG浓度为40、80、160、320、640μg/ml时,上清可溶性血管细胞黏附分子-1浓度分别为(38.51±3.762)对(24.78±250)pg/ml,(61.34±6.99)对(38.20±3.17)pg/ml,(135.60±24.46)对(63.73±5.08)pg/ml,(221.3±29.40)对(112.80±8.91)pg/ml,(420.90±31.70)对(224.40±20.79)pg/ml(P值均<0.01).HUVEC与AECA阳性的IgG共培养后,凝血活性因子表达显著增高:IgG浓度为80、160,320、640 μg/ml时,上清血管性血友病因子浓度分别为(19.51±0.72)对(17.17±0.60) ng/ml,(22.97±1.18)对(18.27±0.614)ng/ml,(26.40± 1.54)对(19.53±0.701)ng/ml,(34.35±1.60)对(23.81±0.92)ng/ml(P值均<0.05);IgG浓度为320、640 μg/ml时,上清血栓调节素浓度分别为(57.50±4.50)对(40.31±4.39)pg/ml,(59.18±4.11)对(38.84±5.16)pg/ml(P值均<0.05).AECA阳性IgG对HUVEC分泌炎性因子(IL-1β、IL-6、IL-8及ANG2)无明显作用(P>0.05).此外,AECA并不诱导HUVEC凋亡,对细胞增殖亦无影响(P>0.05).结论 AECA能引起EC功能发生显著变化,并不引起细胞凋亡,对于阐明AECA介导的EC损伤在移植物抗宿主病等移植并发症中的作用机制有重要意义.
Objective To clarify the role of endothelial cells (ECs) injury induced by antiendothelial cell antibody (AECA) in allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Serum immunoglobulin (IgG) from allo-HSCT recipients were purified and incubated with human umbilical vein vascular endothelium (HUVEC) in vitro,then the functional changes and cell apoptosis were tested.Results After incubation with AECA positive IgG,soluble adhesion molecules significantly elevated in culture supernatant.When concentration of IgG was 160,320,and 640 μg/ml,concentrations of soluble intercellular adhesion molecule-1 in supernatant were statistically higher in AECA positive groups [(117.10±12.82) vs (78.17±4.90) pg/ml,(151.30±15.35) vs (89.46±6.02) pg/ml,(239.00±32.53) vs (127.80± 13.86) pg/ml,P<0.01)].When concentration of IgG was 40,80,160,320,and 640 μg/ml,concentrations of soluble vascular cell adhesion molecule-1 in supernatant were also statistically higher in AECA positive groups [(38.51±3.76) vs (24.78±2.59) pg/ml,(61.34±6.99) vs (38.20±3.17) pg/ml,(135.60±24.46) vs (63.73±5.08) pg/ml,(221.30±29.40) vs (112.80±8.91) pg/ml,(420.90 ± 31.70) vs (224.40 ± 20.79) pg/ml,P<0.01].Clotting activity factors also elevated in culture supematant after incubation with AECA positive IgG.When concentration of IgG was 80,160,320,and 640 μg/ml,concentrations of von Willebrand factor were statistically higher in AECA positive groups [(19.51±0.72) vs (17.17±0.60) ng/ml,P=0.0193;(22.97±1.18) vs (18.27±0.61) ng/ml,(26.40±1.54) vs (19.53±0.70) ng/ml,(34.35±1.60) vs (23.81±0.92) ng/ml,P<0.01].When concentration ofIgG was 320 and 640 μg/ml,concentrations of thrombomodulin were statistically higher in AECA positive groups [(57.50±4.50) vs (40.31±4.39) pg/ml,P=0.0132;(59.18±4.11) vs (38.84±5.16) pg/ml,P<0.01].However,inflammatory factors (IL-1β,IL-6,IL-8 and A