宫内感染HBV免疫失败儿童Th1和Th2型细胞因子mRNA转录研究

Expression of the messenger RNA of type 1 and type 2 cytokines in intrauterus infected hepatitis B immunization failure children

目的 探讨Th1型和Th2型细胞因子mRNA转录变化对宫内感染乙型肝炎病毒(HBV)后免疫失败的影响。方法 采用半定量逆转录 聚合酶链反应 (RT PCR)对 2 9例宫内感染免疫失败儿童、9例宫内感染免疫有效儿童及 2 5例正常免疫儿童外周血单个核细胞在丝裂原 (植物血凝素和细菌脂多糖 )、酵母重组乙型肝炎表面抗原 (HBsAg)及无刺激物时的γ 干扰素 (IFN γ)、白细胞介素 4(IL 4)mRNA表达水平进行检测。结果 IFN γmRNA自发表达在免疫失败组增高 ( 95 %可信限免疫失败组 16 6 5～ 30 89,对照组 76 3～ 16 0 9,P =0 .0 0 8)。与自发表达比较 ,在HBsAg刺激时 ,对照组、有效组和免疫失败组均表现为转录增加 ,但免疫失败组增加值显著低于对照组 (小剂量HBsAg刺激增加值 95 %可信限对照组 899～ 170 3 ,免疫失败组 170～ 85 4,P =0 .0 0 4)。IL 4mRNA自发转录和丝裂原刺激时转录水平在 3组间差异无显著性 ,大剂量HBsAg刺激时对照组表现为转录增加 (增加值 95 %可信限 2 13～ 86 1,P =0 .0 11) ,而免疫失败组无显著增加。丝裂原刺激时IFN γmRNA转录水平和大剂量HBsAg刺激时IFN γmRNA/IL 4mRNA与血清丙氨酸转氨酶水平正相关。 结论 宫内感染HBV免疫失败儿童存在Th1型和Th2型细胞因子的特异性反应低下 ,Th1型细胞因?

Objective To investigate the mRNA response of type1 and type2 cytokines in immunization failure children who were infected hepatitis B virus (HBV) via placenta from HBV carrier mothers. Methods All subjects were born to HBV carrier mothers and received hepatitis B immunoprophylaxis treatment according to routine schedule. Interleukin-4 (IL-4) and interferon-gamma (IFN-γ) transcription of peripheral blood mononuclear cells upon lectins or two-doses of HBsAg from 29 children with persistent HBV infection since birth (Nonresponders), 9 with transient HBV infection at birth and at less than 6 month of age (Responders) were analyzed by semi-quantitative RT-PCR. 25 children with positive anti-HBs response and never positive HBsAg were used as controls. Results Nonresponders expressed a marked higher spontaneous IFN-γ mRNA transcription than controls (95% confidence interval is 1665～3089 in nonresponders,763～1609 in controls, P=0.008). Upon two doses of HBsAg stimulation, all three groups expressed a higher IFN-γ mRNA transcription than spontaneous transcription, but the increase in nonresponders is lower than controls (95% confidence interval by low dose of HBsAg is 899～1703 in controls, 170～854 in nonresponders,P=0.004). The responders expressed a significant increase of IL-4 mRNA transcription upon high-dose HBsAg stimulation but not in nonresponders (95% confidence interval of the increase in controls is 213～861, P=0.011). Positive correlations were found between ALT level and the IFN-γ mRNA transcription upon lectins stimulation and between ALT level and IFN-γ mRNA/IL-4 mRNA ratio upon high-dose HBsAg stimulation.Conclusions Low responses of type 1 and type 2 cytokines transcription upon specific stimulation exist in intrauteral infected hepatitis B immunization failure children, and the over response of type 1 cytokine upon nonspecific stimulation may contribute to the liver injury in nonresponders.