乙型肝炎病毒感染者血清和外周血单个核细胞培养上清细胞因子测定及临床意义

The Study of Cytokine Level in Sera and Culture Medium of Peripheral Blood Mononuclear Cells in Patients with Hepatitis B Virus Infection

目的观察不同HBV感染者血清和外周血单个核细胞(PBMC)培养上清中的细胞因子水平。方法采集不同HBV感染者血清;制备PBMC并用rHBcAg刺激,培养72h后收集培养上清。采用酶联免疫吸附法检测血清和培养上清中的IFN-γ和IL-4水平。结果血清IFN-γ水平,急性乙肝显著高于健康对照和HBV携带者(P<0.01和P<0.05),也高于慢性乙肝;慢性乙肝显著高于健康对照(P<0.05)。血清IL-4水平,急、慢性乙肝均显著高于健康对照和HBV携带者(P<0.01和P<0.05),HBV携带者与健康对照相比差异无统计学意义。血清IFN-γ/IL-4比值,除慢性乙肝显著低于健康对照(P<0.01),其余组间差异无统计学意义。在PBMC培养上清中,急、慢性乙肝患者的IFN-γ水平显著高于健康对照和HBV携带者(P<0.05),急性肝炎组高于慢性组,但差异无统计学意义;HBV携带者与健康对照相比,差异无统计学意义。3个HBV感染组的IL-4水平与健康对照相比,差异无统计学意义。结论 (1)与使用rHBcAg刺激后的PBMC培养上清相比,血清检测可能更加符合患者的真实细胞因子水平。(2)HBV感染时IFN-γ表达水平和IFN-γ/IL-4比值与疾病转归密切相关。

Objective To investigate cytokine levels in serum and culture medium of peripheral blood mononuclear cells （PBMC） from patients with hepatitis B virus（HBV） infection. Methods PBMCs isolated from fresh heparinized blood were cultured and stimulated with rHBcAg. After 72h at 37℃5% CO2 in air, the culture supernatant was collected. Levels of interferon - gamma （ IFN - γ） and interleukin （IL） -4 in blood serum in spontaneous and supernatant were measured by enzyme -linked immunosorbent assay （ELISA）. Resuits Serum IFN - γ levels in patients with acute self - limited hepatitis B （AH） and chronic hapetiti B （CHB） were significantly higher than those in normal control（NC） （P 〈 0. 01 and P 〈 0.05 ）, but there was signifieent difference between two HBV infection groups. Serum IL - 4 levels in AH and CHB were significantly higher those in NC （ P 〈 0.01 ） and asymptomatic HBV carriers （ P 〈 O. 05 ）. IFN - 3＂/ IL - 4 ratio in AH was closed to that in NC, but IFN - γ/ IL - 4 ratio in CHB showed significant decrease（ P 〈 0. 01 ）. CHB groups showed significant decrease in IFN - γ/IL -4 ratio than that in NC. IFN - γ/IL -4 ratio had no significant differences between every two groups, except CHB. Compared with NC and HBV carriers, AH and CHB showed significant increase in supernatant IFN -γ levels, but three HBV infection groups showed no significant changes in supernatant IL - 4 levels. Conclusion （ 1 ） Compared with culture supernatant obtained from stimulated with rHBcAg, spontaneous serum cytokine levels may be closer to real immune responses. （2）The elevated IFN -γ level and immune balance of cytokine profile exert a profound influence on the prognosis of HBV infections.