尖锐湿疣患者外周血自然杀伤细胞活性的研究

目的　从自然杀伤细胞 (NK)内分泌γ 干扰素和CD6 9表达等层次 ,探讨尖锐湿疣 (CA)患者的细胞免疫状态 ,为临床免疫治疗提供指引。方法　用密度梯度离心法分离出外周血单个核细胞 (PBMC) ,进行细胞培养和免疫荧光染色 :①PDB刺激PBMC ,2h、12h、2 4h后收获细胞 ,进行anti CD5 6 FITC与anti CD6 9 PE免疫标记 ;②PDB、Ionomycin和Monensin刺激PBMC ,4h收获细胞 ,用anti CD5 6 PE与anti IFN γ FITC细胞内染色 ,最后利用流式细胞仪对标记细胞进行分析。结果　NK细胞受体外刺激后 ,长病程组各时点的CD6 9阳性细胞百分率分别是 ( 2 5 .73± 2 .99) %、( 5 7.0 8±4.74) %、( 6 7.0 7± 2 .5 2 ) % ,与对照组各时点相比较 ,差异均有显著性 (P <0 .0 5 ) ,而短病程组各时点与对照组比较 ,差异均无显著性 (P >0 .0 5 )。长病程组的IFN γ阳性NK细胞比率是 ( 34.6 9± 3 .36 ) % ,与对照组比较差异有显著性 (P <0 .0 5 )。短病程组与对照组比较差异无显著性 (P >0 .0 5 )。结论　短病程患者NK细胞活化和功能均正常 ,但长病程者NK细胞活化明显不足和功能严重障碍。本试验结果为长病程患者的免疫治疗提供理论依据和新的指引。

Studies on the Activation of NK Cell from Human Peripheral Blood of the Patients with Condyloma Acuminate

Objective To improve the clinical therapy,studies were performed on the details of intracellular IFN-γ secretion and CD69 expression on NK cell in vitro for investigating the statute of cellular immunity of the patients with condyloma acuminate(CA).Methods After isolated the PBMC from anti-clotting human peripheral blood with heparin by Ficoll-Hypaque centrifugation,the PBMC culture procedure in vitro and two-color immunofluorescent staining were undertaken as following:①PBMC were activated 2,12,24 hours by PDB,then stained with anti-CD56-FITC and anti-CD69-PE.②PBMC were stimulated 4 hours by PDB,Ionomycinand and Monensin together,then intracellular stained with anti-CD56-PE and anti-IFN-γ-FITC.At last the stained cells were analyzed by flow cytometry.Results The percentages of NK-cells expressing CD69 of long history-CA group at 2,12,24 hour were(25.73±2.99)%,(57.08±4.74)%,(67.07±2.52)% respectively,the differences between them and those of control group were significant(P<0.05)But the differences between the short history-CA group and those of control group were no significant(P>0.05),The ratios of IFN-γ positive NK-cells of long history-CA group were (34.69±3.36)%,the difference between them and control group was significant(P<0.05).But there were no difference between the short history-CA group and those of control group (P>0.05).Conclusion The activation and function of NK cells of the patients with short history-CA were normal,but those of the patients of long history-CA were deficient.These data would provide the theory and new ways for the immune therapy of especially long history-CA.