慢性乙型病毒感染者树突状细胞体外经HBsAg活化后的免疫效应

Immune effect of dendritic cell activated by HBsAg in patients with chronic HBV infective

目的研究慢性乙型病毒感染者与健康人外周血树突状细胞（DC）经HBsAg活化后的免疫功能的差异。方法从慢性HBV感染者、健康人外周血中培养扩增DC和CIK，在DC成熟前，加入纯的HBsAg刺激，并再与同一来源的CIK共同培养。用流式细胞仪检测DC、CIK表型，用ELISA法检测DC、CIK共培养上清液中的IL-12浓度，用CCK．8比色法测定DC诱导CIK对HepG2．2．15细胞的杀伤活性。结果未经HBsAg致敏的健康人DC表面标志CDla、CD80及CD83明显高于慢性HBV感染者（P〈0．05）；经HBsAg致敏的健康人DC表面标志均高于慢性HBV‘感染者，差异有统计学意义（P〈0．01）；慢性HBV感染者经HBsAg致敏的DC表面标志与未经HBsAg致敏者比较差异无统计学意义（P〉0．05）。CIK细胞CD3CD8、CD3CD56的双阳性表达率健康人HBsAg致敏者明显高于慢性HBV感染者及未经HBsAg致敏的健康人（P〈0．01，P〈0．05）；慢性HBV感染组HBsAg致敏与未经HBsAg致敏者比较差异无统计学意义（P〉0．05）。HBsAg致敏DC诱导CIK对HepG2．2．15细胞的杀伤率健康人显著高于慢性HBV感染者（P〈0．01）。健康人HBsAg致敏DC、CIK共培养上清液中的IL-12浓度显著高于慢性HBV感染者（P〈0．01）；慢性HBV感染者HBsAg致敏与未经HBsAg致敏IL-12含量差异无统计学意义（P〉0．05）。结论慢性乙型病毒感染者、健康人DC经HBsAg活化后对HepG2．2．15细胞的免疫效应存在显著差异：健康人高于慢性HBV感染者。

Objective To study differences of immune effect of dendritic cell activated by HB- sag between patients with chronic HBV and without HBV infective in vitro. Methods Dendritic cells （DC） and cytokine-induced killer（CIK）cells were cultured and amplified from chronic HBV infection and human peripheral blood . Before the DC mature, it was stimulation with adding pure HBsAg in cell culture medium. DC and CIK phenotype were detected by flow cytometry method. ELISA was used to de- tect the level of IL-12 in the upper liquid medium which DC and CIK cell mixed culture. The cell-killing activity of DC-induced CIK cell for HepG2.2. 15 was measured by using CCK-8 colorimetric assay. Re-suits Except HLA-DR,the positive rate of DC surface marker CDla, CD50 and CD83 in healthy people were higher than in chronic HBV infection（ P 〈0. 05）,but healthy peopleDC activated by HBsAg, four surface markers were significantly higher than in chronic HBV infection （P 〈 0. 01 ）. With or without HBsAg-pulsed ,there was no significant difference in DC surface markers of chronic HBV infection. CD3 CD8, CD3 CD56 double positive expression rate of CIK cells in healthy people with HBsAg-pulsed were higher than in chronic HBV infection and those healthy without with HB- sAg-pulsed. The difference statistically significant （ P 〈 0. 01, P 〈 0. 05 ）. There were not significantly differences chronic in HBV infection with or without HBsAg-pulsed （ P 〉 0.05 ） ; At healthy people, CIK cells HBsAg pulsed DC induced cell-killing rate of HepG2.2. 15 ceils was significantly higher than those in the chronic HBV infection （ P 〈0. 01 ）. The level of IL-12 in HBsAg pulsed DC-CIK co-culture supermatants of healthy people was much higher than that of chronic HBV infection （ P 〈 0. 001 ）. There were not significant differences in chronic HBV infection with HBsAg vs without HBsAg pulsed （ P 〉 0. 05 ）. Conclusions The activation of the immune effect of HBsAg pulsed for HepG2. 2. 15 cells were sig- nificantly different ：the healthy is stronger than that chronic HBV infection＇ s.