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目的确认重组人白细胞介素12(rhIL-12)联合乙型肝炎表面抗原(HBsAg)可增强慢性乙型肝炎患者外周血单个核细胞(PBMC)对HBsAg产生特异性细胞免疫应答,探讨rhIL-12联合HBsAg作为治疗型乙型肝炎疫苗的应用前景。方法取8名正常人和16例慢性乙型肝炎患者的PBMC,分别用不同浓度的HBsAg、rhIL-12单独体外刺激,再用HBsAg联合rhIL-12(0.01、0.1和1.0ng/mL)刺激患者PBMC,72h后用酶联免疫吸附试验(ELISA)测定培养上清中的γ干扰素(IFN-γ)。结果HBsAg或rhIL-12单独刺激仅产生低水平的IFN-γ,联合使用HBsAg和rhIL-12刺激后,患者PBMC产生的IFN-γ平均水平显著高于单独使用HBsAg或rhIL-12(P均<0.05)。HBsAg(0.5μg/mL)联合rhIL-12(0.01、0.1和1.0ng/mL)刺激患者PBMC产生的IFN-γ与rhIL-12的浓度之间呈明显的量效关系。结论HBsAg和rhIL-12联合刺激乙型肝炎患者PBMC,通过产生高水平IFN-γ激活乙型肝炎患者特异性细胞免疫。
Objective To confirm that recombinant human interleukin-12 (rhIL-12) combined with hepatitis B surface antigen (HBsAg) can enhance the specific cellular immune response to HBsAg in peripheral blood mononuclear cells (PBMCs) from patients with chronic hepatitis B and to explore the effect of rhIL-12 Application prospect of HBsAg as a therapeutic hepatitis B vaccine. Methods Eight PBMCs from normal subjects and 16 patients with chronic hepatitis B were stimulated with HBsAg and rhIL-12 at different concentrations in vitro and then stimulated with HBsAg and rhIL-12 (0.01, 0.1 and 1.0 ng / mL) respectively After 72h, IFN-γ in the culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA). Results HBsAg or rhIL-12 stimulation alone produced only low levels of IFN-γ. The combined IFN-γ production of PBMC in patients with HBsAg and rhIL-12 stimulation was significantly higher than that of HBsAg or rhIL-12 alone (all P < 0.05). HBsAg (0.5μg / mL) combined with rhIL-12 (0.01,0.1 and 1.0ng / mL) in patients with PBMC produced IFN-γ and rhIL-12 concentration showed a significant dose-effect relationship. Conclusion The combination of HBsAg and rhIL-12 stimulates PBMC in patients with hepatitis B and activates specific cellular immunity in patients with hepatitis B by generating high levels of IFN-γ.