自制改进溶血素用于慢加急性肝衰竭患者免疫细胞的流式检测  

Detection of immune cells in patients with acute-on-chronic liver failure by flow cytometry with home-made lysis solution

作　　者：周春保[1] 徐向升[1] 徐若男[1] 袁今虹[1] 金磊[1] 张政[1] 王福生[1] 

机构地区：[1]中国人民解放军第三○二医院肝病生物治疗研究中心,北京100039

出　　处：《中国病毒病杂志》2013年第2期117-121,共5页Chinese Journal of Viral Diseases

基　　金：国家自然科学基金(81172779)

摘　　要：目的探讨自制改进溶血素与BD LysingTMSolution对健康人(HC)、慢性乙型肝炎(CHB)和慢加急性肝衰竭(ACLF)患者外周血淋巴细胞、单核细胞形态、比例变化和表面标志的影响。方法用两组溶血素(自制改进溶血素和BD LysingTMSolution)处理10例HC、20例CHB和20例ACLF患者的抗凝外周血,观察两种溶血素对红细胞裂解的影响,同时应用流式细胞仪检测淋巴细胞、单核细胞亚群比例,并观察溶血素对其影响。结果改进溶血素和BD LysingTMSolution均能彻底裂解红细胞,两种溶血素处理的淋巴细胞、单核细胞、中性粒细胞在流式细胞仪上分群清晰;在HC和CHB患者,单核细胞和淋巴细胞比例差异无统计学意义;在ACLF患者,改进溶血素和BD LysingTMSolution溶血后获得的淋巴细胞比例差异无统计学意义,但改进溶血素溶血后获得的单核细胞比例却显著高于BD LysingTMSolution溶血后所获得的比例(P<0.05)。结论在流式细胞仪检测淋巴细胞时,改进溶血素和BD LysingTMSolution效果相同;但在ACLF患者进行单核细胞研究时,改进溶血素明显优于BD LysingTMSolution,且可明显降低研究成本。Objective To compare the results of our home-made lysis solution and the commercial solution （BD LysingTM Solution） on the morphology and the proportion of peripheral blood lymphocytes and monocytes in healthy controls （HC）, patients with chronic hepatitis B （CHB） and acute-on-chronic liver failure （ACLF）. Methods A total of 10 HC subjects, 20 CHB and 20 ACLF patients were enrolled in the study. Peripheral blood was lysed by using both home-made lysis solution and BD LysingTM Solution. The lysis results were ana- lyzed by flowcytometry to compare the proportion of lymphocytes and monocytes. Results Our home-made ly- sis solution showed similar effects when comparing with the BD LysingTM Solution in red blood cell lysis; no significant damage on the lymphocytes, monocytes and neutrophils was found comparing to lAD LysingTM Solu- tion. The proportion of lymphocyte was also similar with two solutions for both HC subjects and CHB patients （P〉0.05）; nevertheless, the proportion of monocytes treated by home-made lysis buffer is significantly higher than that acquired by using the BD LysingTM Solution （P〈0.05） . Conclusions Our home-made lysis solution showed similar effects to the BD LysingTM Solution in the study of lymphocytes and better performance for monocytes in cytometry analysis.

关 键 词：溶血素 BD LysingTM Solution 单核细胞 淋巴细胞 慢加急性肝衰竭 流式细胞术 

分 类 号：R575.3[医药卫生—消化系统]