HBV感染对体外培养NK细胞生物活性影响的研究

Effect of HBV infection on biological activity of NK cells cultured in vitro

目的HBV感染对NK细胞生物活性的影响。方法外周血标本来源于2018年1月至2018年12月在中山大学附属第三医院岭南医院经体外扩增培养NK细胞后回输的49例患者。患者均签署知情同意书,符合医学伦理学规定。其中男30例,女19例;年龄27~87岁,中位年龄60岁。根据患者有无HBV感染,将血标本分为HBV(+)组和HBV(-)组。体外扩增、培养CD3^-CD56^+的NK细胞和CD3^+CD56^+的NKT细胞。采用流式细胞术检测NK、NKT细胞百分率。两组细胞活性率和细胞凋亡等比较采用t检验。结果HBV(-)组细胞活性率为(94.4±1.2)%,明显高于HBV(+)组的(85.1±4.6)%(t=2.146,P<0.05);而HBV(-)组细胞凋亡率为(3.2±0.9)%,明显低于HBV(+)组的(11.6±4.1)%(t=-2.220,P<0.05)。HBV(-)组NK细胞百分率为(55.8±4.4)%,明显高于HBV(+)组的(34.7±5.9)%(t=2.889,P<0.05)。结论HBV(-)患者来源的细胞体外扩增诱导可获得较多NK细胞。

Objective To investigate the effect of HBV infection on the biological activity of NK cells.Methods Peripheral blood samples were collected from 49 patients who received reinfusion of NK cells cultured in vitro Lingnan Hospital of the Third Affiliated Hospital of Sun Yat-sen University from January 2018 to December 2018.The informed consents of all patients were obtained and the local ethical committee approval was received.Among them,30 patients were male and 19 female,aged 27-87 years old with a median age of 60 years old.Blood samples were divided into the HBV(+)and HBV(-)groups according to the incidence of HBV infection.CD3^-CD56^+NK cells and CD3^+CD56^+NKT cells were proliferated and cultured in vitro.The percentage of NK and NKT cells was detected by flow cytometry.The cell activity rate and apoptosis rate were statistically compared between two groups by t test.Results The cell activity rate in the HBV(-)group was(94.4±1.2)%,significantly higher than(85.1±4.6)%in the HBV(+)group(t=2.146,P<0.05).The apoptosis rate in the HBV(-)group was(3.2±0.9)%,significantly lower than(11.6±4.1)%in the HBV(+)group(t=-2.220,P<0.05).The percentage of NK cells in the HBV(-)group was(55.8±4.4)%,significantly higher than(34.7±5.9)%in the HBV(+)group(t=2.889,P<0.05).Conclusions More NK cells can be obtained from HBV(-)patient-derived cells proliferated and cultured in vitro.