外周血不同制备法对检测两种性病患者淋巴细胞免疫表型的影响

Study on the lymphocyte immunophenotyping using different peripheral blood cell preparations in the patients with condyloma acuminatum and recurrent genital herpes

目的:探讨流式细胞术中的裂解全血法和分离淋巴细胞法对检测尖锐湿疣(CA)和复发性生殖器疱疹(RGH)患者外周血淋巴细胞免疫表型实验结果的影响。方法:采用单色、双色和三色荧光抗体染色技术经流式细胞仪分别检测20例RGH、CA患者的裂解全血及10例RGH、CA患者的分离淋巴细胞中的CD3+T细胞、CD4+T细胞、CD8+T细胞、CD40抗原、CD40L抗原、T细胞αβ链(Tαβ)和T细胞γδ链(Tγδ)染色阳性细胞百分率。结果:裂解全血法和分离淋巴细胞法分析淋巴细胞免疫表型结果,除CD40L(P=0.89)没有显著性差异以外,其余均有显著性差异(分别为CD3+T:P<0.01,CD4+T:P<0.05,CD8+T:P<0.05,CD40:P<0.05,Tαβ:P<0.001,Tγδ:P<0.05)。结论:CA和RGH患者外周血不同制备方法影响淋巴细胞免疫表型结果分析,裂解全血法优于分离淋巴细胞法。

Objective： To explore the effect of lysed whole blood （WBL） method and Ficoll - Pague separated peripheral blood lymphocyte （ F - PBL） method using flow cytometry on immunophenotyping in peripheral blood lymphocyte of patients with condyloma acuminatum （CA） and recurrent genital herpes （RGH）. Methods： Single- color, two- color and three- color immunofluorescent staining of cell surface antigens （CD3^＋ , CD8^＋ , CD4^＋ , CD40, CD40L, T cell αβ chains, T cell γδ chains） from 20 patients of CA and BGH by WBL method and 10 patients of CA and RGH by F- PBL method with flow cytometric analysis were performed. Results： The lymphocyte immunophenotyping measured by WBL method and F - PBL method was markedly different （CD^3＋ T： P 〈 0.01, CD4^＋T：P〈0.05,CD8^＋T：P〈0.05,CD40：P〈0.05,Tαβ：P〈0.01,Tγδ：P〈0.05, respectively）, except for CD40L （ P - 0.89）. Conclusion： Peripheral blood cell preparation of patients with CA and RGH influences the levels of the lymphocyte immunophenotyping. The WBL method is better than the F- PBL method.