特发性血小板减少性紫癜患者免疫相关指标的检测及其临床意义

目的 检测特发性血小板减少性紫癜(ITP)患者免疫相关指标的变化,探讨其在ITP发病机制中的作用及其临床意义.方法 应用酶联免疫斑点技术(ELISPOT)、改良血小板抗原单克隆抗体固相化检测技术(MAIPA)、流式细胞术及夹心法ELISA分别检测64例1TP患者及31例正常对照者分泌GPⅡb/Ⅲa抗体B细胞、血小板特异性抗体(抗GPⅡb/Ⅲa抗体、抗GP I b/Ⅸ抗体)、T淋巴细胞亚群、网织血小板(RP)及血小板生成素(TPO)的变化.结果 ITP患者分泌GPⅡb/Ⅲa抗体B细胞频数急性ITP组患者为7.6±4.6/105个外周血单个核细胞(PBMC),慢性ITP组患者为5.3±3.0/105个PBMC]、血小板特异性抗体(抗GPⅡb/Ⅲa抗体、抗GPI b/Ⅸ抗体)的吸光度值(急性ITP组患者为0.51±0.11、0.48±0.06,慢性ITP组患者为0.49±0.10、0.46±0.09)、CD8+T淋巴细胞百分比(27.09±9.86)%]、RP百分比巨核细胞增多组为(24.85±19.18)%,巨核细胞正常组为(23.89±18.90)%]明显高于正常对照组1.3±0.5/105个PBMC,0.33±0.06,0.41±0.03,(22.08±4 54)%,(8.19±2.46)%,P值均＜0.05],其中急性ITP患者分泌GPⅡb/Ⅲa抗体B细胞高于慢性ITP患者(P＜0.05).ITP组CD3+T淋巴细胞百分比、CD4+T淋巴细胞百分比及CD4+/CD8+比值(60.88±14.59)%、(28.41±10.55)%、1.18±0.59]均低于正常对照组(69.89±6.43)%、(35.38±5.05)%、1.64±0.29,P值均＜0.05].ITP患者巨核细胞增多组TPO水平(72.09±41.64)明显低于ITP患者巨核细胞正常组(118.60±70.72,P＜0.05),与正常对照组(75.37±26.32)之间差异无统计学意义(P＞0.05).结论 分泌GPⅡb/Ⅲa抗体B细胞、血小板特异性抗体、T淋巴细胞亚群、RP%及TPO在ITP诊断及指导定向干预治疗中有一定的意义.

The clinical significance of immune-related marker detection in idiopathic thrombocytopenic purpura

Objective To assess the clinical significance of detecting the immune markers in idiopathic thrombocytopenic purpura (ITP). Methods The frequencies of circulating B cells secreting platelet-specific antibody, platelet-specific antibody, the percentage of T lymphocyte subsets, the percentage of reticulated platelet and the level of thrombopoietin in 64 ITP patients and 31 healthy controls were measured with enzyme-linked immunospot assay (ELISPOT),modified monoclonal antibody immunobilization of platelet antigens assay (MAIPA), flow cytometry and sandwich enzyme-linked innnunosorbent assay respectively. Results Compared with the controls1.3 ± 0. 5/105 peripheral blood mononuclear cell (PBMC), (0.33±0.06,0.41±0.03), (22.08±4.54)% and (8.19±2.46)%], the frequencies of circulating B cells secreting platelet-specific antibody (7.6±4.6/105 PBMC in acute ITP group, 5.3±3.0/105 PBMC in chronic ITP group), platelet-specific antibody (including the anti-GP Ⅱ b/Ⅲa antibody, anti-GP Ⅰ b/X antibody) (0.51 ±0.11, 0.48±0.06 in acute ITP group; 0.49±0.10,0.46±0.09 in chronic ITP group), the percentage of CD8+ T Lymphocyte (27.09±9.86 ) %, the percentage of reticulated platelet in ITP patientsthe megakaryocyte cytosis group (24. 85 ± 19. 18)%, the normal megakaryocyte group (23.89±18.90)%]were significantly increased ( all P＜0.05).The frequencies of circulating B cells secreting platelet-specific antibody in acute ITP patients were notably increased (P＜0.05) compared to the chronic ITP patients. In T lymphocyte subsets, the percentage of CD3+T lymphocyte and CD4+ T lymphocyte and the ratio of CD4+/CD8+ in the patients with ITP(60.88±14.59)%, (28.41±10.55)%, 1.18±0.59]were notably decreased than those in the healthy controls (69.89±6.43)%, (35.38±5.05) %, 1.64±0.29, P＜0.05]. There was no apparent difference of the level of thrombopoietin between ITP patients with megakaryocyte cytosis (72. 09 ± 41.64 ) and health controls (75.37± 26. 32, P ＞ 0. 05 ), however, the level of thrombopoietin of ITP patients with normal megakaryocyte apparently increased (118.60±70.72, P＜0.05). Conclusion Detecting the frequencies of circulating B cells secreting platelet-specific antibody, platelet-specific antibody, the percentage of T lymphocyte subsets, the percentage of reticulated platelet and the level of thrombepoietin in the patients with ITP may improve the diagnosis and guide clinical therapy.