Sun Ying, Chen Huan-yong, Wang Fei, Zhang Xin, Jiang Hong-qi, Shao Feng-juan, Zhu Si-he
Department of Infectious Diseases, First Clinical Hospital, Harbin Medical University, Harbin 150001, China.
Zhonghua Gan Zang Bing Za Zhi. 2003 Aug;11(8):470-3.
To explore the effect of IL-18 on peripheral blood monocytes (PBMCs) from chronic hepatitis B (CHB) patients and HBV DNA released by HepG2.2.15 cells, which were transfected with the gene of HBV.
PBMCs were isolated from 25 healthy persons and 25 CHB patients, which were co-cultured with HBcAg and IL-18 at different concentrations for 72 hours. The level of IFN-gamma in the culture supernatant of PBMCs was determined by ELISA. One patient' PBMCs were co-cultured for 96 hours with various concentrations of IL-18 and HepG2.2.15 cells which had been cultured for 24 hours, the supernatant was collected to detect HBV DNA level by PCR.
When PBMCs were stimulated by HBcAg and IL-18 at various concentrations, the levels of supernatant IFN-gamma in the CHB group were much higher than those in the normal control group (at 0.2ng/ml: t=11.7, P<0.01; at 1.0ng/ml: t=16.19, P<0.01; at 5.0ng/ml: t=20.12, P<0.01), especially when the PBMCs were stimulated by HBcAg, IL-18 and IL-12 (1313.20pg/ml+-187.76pg/ml vs. 390.75pg/ml+-43.23pg/ml, t=23.94, P<0.01). The IFN-gamma level in the patients who were stimulated by HBcAg alone was much lower than the levels in the patients who were stimulated by HBcAg and IL-18 at various concentrations, and which were lower than those in the patients stimulated by HBcAg, IL-12 and IL-18 at the same concentrations (light: t=2.2, P<0.05; moderate: t=2.97, P<0.05). The HBV DNA content in the supernatant of co-cultivation with HepG2.2.15 cells and PBMCs was much higher than that of the two kinds of cells stimulated by HBcAg and IL-18 at various concentrations or HBcAg, IL-18 and IL-12/IFN-a1b.
IL-18 can improve the PBMCs from CHB patients to produce a great deal of IFN-gamma, so it has a good application prospect in two aspects: immunoregulatory effects and increasing the ability to kill the cells infected with virus.
探讨白细胞介素-18(IL-18)对慢性乙型肝炎(CHB)患者外周血单个核细胞(PBMCs)及转染乙肝病毒(HBV)基因的HepG2.2.15细胞释放HBV DNA的影响。
从25名健康人和25名CHB患者中分离PBMCs,将其与不同浓度的乙肝核心抗原(HBcAg)和IL-18共培养72小时。采用酶联免疫吸附测定法(ELISA)检测PBMCs培养上清液中γ干扰素(IFN-γ)水平。将1例患者的PBMCs与不同浓度的IL-18及已培养24小时的HepG2.2.15细胞共培养96小时,收集上清液,采用聚合酶链反应(PCR)检测HBV DNA水平。
不同浓度的HBcAg和IL-18刺激PBMCs时,CHB组上清液IFN-γ水平明显高于正常对照组(0.2ng/ml时:t=11.7,P<0.01;1.0ng/ml时:t=16.19,P<0.01;5.0ng/ml时:t=20.12,P<0.01),尤其是HBcAg、IL-18和IL-12共同刺激时(1313.20pg/ml±187.76pg/ml对390.75pg/ml±43.23pg/ml,t=23.94,P<0.01)。单独HBcAg刺激患者的IFN-γ水平明显低于不同浓度HBcAg和IL-18刺激患者的水平,且低于相同浓度HBcAg、IL-12和IL-18刺激患者的水平(轻度:t=2.2,P<0.05;中度:t=2.97,P<0.05)。HepG2.2.15细胞与PBMCs共培养上清液中的HBV DNA含量明显高于不同浓度HBcAg和IL-18或HBcAg、IL-18和IL-12/干扰素α1b刺激的两种细胞。
IL-18能促进CHB患者PBMCs产生大量IFN-γ,在免疫调节及提高杀伤病毒感染细胞能力两方面均具有良好的应用前景。