细胞因子诱导的杀伤细胞及上清液对肺炎克雷伯杆菌的作用

Cytokine-induced killer cells and supernatant on the role of klebsiella pneumoniae

目的 观察细胞因子诱导的杀伤细胞（CIK）及CIK细胞培养上清液对肺炎克雷伯杆菌的体外抑菌作用.方法 体外诱导制备CIK细胞,流式细胞术分析第1、7、14、21天CIK细胞表型,微量肉汤稀释法检测CIK细胞对肺炎克雷伯杆菌（细菌接种后浓度为1.0×105 cfu/ml,效靶比为40∶1、20∶1、10∶1、5∶1）、上清液、及联合浓度梯度的阿米卡星（AK）、哌拉西林（PIPC）后对菌株的最低抑菌浓度（MIC）值,确定其体外抗菌活性.结果 CD3＋ CD56＋细胞于培养第14 ～21天时所占比例维持在45％以上;吸光度（A）值检测结果示24 h时各浓度CIK细胞对细菌杀伤作用差异无统计学意义（F =3.065,P＞0.05）;上清液组未见明显细菌生长;AK、PIPC的MIC值分别为2 mg/L和512 mg/L;CIK细胞联合AK、PIPC未降低后两者的MIC值,但上清液联合AK、PIPC后,两者的MIC值分别降至最低接种浓度0.25 mg/L和64.00 mg/L以下.结论 CIK细胞对肺炎克雷伯杆菌无抗菌活性,且联合抗生素后无增效反应,但其上清液具有体外抑菌作用.

Objective To investigate the ability of cytokine-induced killer （CIK） cells and supernatant against klebsiella pneumoniae in vitro.Methods CIK cells were prepared in vitro.The phenotype of CIK cells was analyzed by flow cytometer in 1,7,14,21 day.Both microdilution method was used to detect the minimum inhibitory concentration （MIC） values of CIK cells against klebsiella pneumoniae （final inoculation concentration of bacteria was 1.0 × 105 cfu/ml,target ratio was 40∶ 1,20∶ 1,10∶ 1,5∶1）,the supernatant,and amikacin （AK）,piperacillin （PIPC） joint after,to determine their antimicrobial activity in vitro.Results In the culture section 14-21 day,CD3 ＋ CD56 ＋ cell phenotype proportion remained at a high level more than 45%.Absorbance （A） values for each test showed that different concentrations of CIK cells killing effect were no statistically significant （F =3.065,P 〉 0.05） at 24 h.The supernatant group showed no significant bacterial growth.The MIC values of amikacin （AK）,piperacillin （PIPC） were 2,512 mg/L respectively.CIK cells respectively,and AK,PIPC joint after,the latter failed to reduce the MIC values,but the supernatant reduced the MIC values of both to the minimize inoculum concentrations of 0.25 mg/L and 64.00 mg/L.Conclusion CIK cells showed no direct killing effect against klebsiella pneumoniae in vitro,and no synergistic reaction combined with antibiotics,but the supernatant has antibacterial effects to klebsiella pneumoniae.