慢性HBV感染者、健康人树突状细胞经HBsAg活化后的免疫效应差异

目的:研究慢性HBV感染者、健康人外周血树突状细胞(dendriticcells,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,差异具有统计学意义(P<0.01);慢性HBV感染者经HBsAg致敏的DC表面标志与未经HBsAg致敏无显著性差异.CIK细胞CD3、CD8、CD3、CD56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致敏者比较无显著性差异(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<0.001);慢性HBV感染者HBsAg致敏与未经HBsAg致敏IL-12含量差异不显著(P>0.05).结论:慢性HBV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.

Difference in immune effect of dendritic cells activated by HBsAg between patients with chronic HBV infection and healthy people

AIM:To compare the difference in immune effect of dendritic cells activated by HBsAg in patients with chronic HBV and healthy people.METHODS:Dendritic cells(DCs) and cytokineinduced killer(CIK) cells were cultured and amplified from the peripheral blood of patients with chronic HBV infection and healthy people.DCs were stimulated with pure HBsAg in cell culture medium prior to maturation.DC and CIK phenotypes were detected by flow cytometry.ELISA was used to detect the level of IL-12in the supernatants of co-cultured DCs and CIK cells.The cell-killing activity of DC-induced CIK cells against HepG2.2.15 cells was measured using CCK-8 colorimetric assay.RESULTS:Except HLA-DR,the positive rates of DC surface markers Cdla,CD80,and CD83 in DCs from healthy people were higher than those in DCs from patients with chronic HBV infection(all P < 0.05),but the positive rates of all four surface markers in healthy people DCs activated by HBsAg were significantly higher than in DCs from patients with chronic HBV infection(P < 0.01).There was no significant difference in the positive rates of DC surface markers between HBsAg-pulsed and non-pulsed DCs from patients with chronic HBV infection.The percentage of CD3+/CD8+ or CD3+/CD56+ CIK cells in HBsAg-pulsed healthy people was significantly higher than those in patients with chronic HBV infection and non-pulsed healthy people(P < 0.01,P < 0.05),but there was no significant difference between HBsAg-pulsed and non-pulsed patients with chronic HBV infection(P > 0.05).The cell-killing activity of HBsAg-pulsed DCinduced CIK cells against HepG2.2.15 cells was significantly higher in healthy people than in patients with chronic HBV infection(P < 0.01).The level of IL-12 in supernatants of co-cultured HBsAg-pulsed DC-CIK cells from healthy people was much higher than that from patients with chronic HBV infection(P < 0.001),but there was no significant difference between HBsAg-pulsed and non-pulsed DCs from patients with chronic HBV infection(P > 0.05).CONCLUSION:The immune effect of HBsAgpulsed DCs for HepG2.2.15 cells was significantly stronger in healthy people than in patients with chronic HBV infection.