乙肝病毒携带者外周血单个核细胞培养上清液HBV-DNA定量检测意义

The Study on the Levels of HBV-DNA in Culture Supernatants of Peripheral Blood Mononuclear Cells From HBeAg Positive Carriers

目的 探讨慢性无症状乙肝病毒携带者外周血单个核细胞 (PBMC)与HBV -DNA定量的临床关系及意义。方法 采用慢性乙肝病毒HBeAg阳性携带者PBMC体外刺激诱导培养 ,荧光定量PCR(FQ -PCR)法测定上清液HBV -DNA ,ELISA法检测白细胞介素 10、12 (IL -10、12 )与γ 干扰素 (γ IFN)。同期检测肝功能 ,血清HBV -DNA定性试验。结果 PBMC培养后于上清液中以FQ -PCR法检出HBV -DNA。 >5 0 0 0考贝 /ml组中 ,IL -12及γ IFN均明显高于正常对照组。重症肝炎PBMC上清液FQ -PCR测定HBV -DNA阳性率明显高于同期血清PCR定性试验阳性率。PBMC上清液FQ -PCR测定HBV -DNA阳性率与血清HBeAg转换的关系同血清HBV -DNA定性试验阳性率与其关系相仿。与肝功能ALT及TBiL程度无明显相关。结论 HBeAg携带者PBMC培养液HBV -DNA高复制状态时IL -12及γ IFN分泌增强。重症肝炎PBMC上清液HBV -DNAFQ

Objective To probe into significanec of HBV-DNA level in cultured supernatants of peripheral blood mononuclear cell(PBMC) from asymptomatic carriers with chronic HBeAg positive.Methods The PBMC from 51 carries of HBeAg positive was cultured by stimuli-induction in vitro.The levels of HBV-DNA were measured by means of fluorescence quantitative PCR(FQ-PCR)in supernatants,the interleukin-10,-12(IL-10,-12) and γ interferon(γ IFN)were also detected by using ELISA.Simultaneously,liver functions were tested and qualitative test was used to assay the HBV-DNA in sere.The levels of HBV-DNA positive in supernatants of PBMC were detected by FQ-PCR after incubation.Results IL-112 and γ IFN in the group of >5000 copy/ml were both significantly higher than those in the controls.The levels of HBV-DNA positive in cultured supernatants of PBMC from suffering grave hepatitis B were higher than those in the sera by qualitative PCR of the same period.They were similar between the positive rate of quantitative PCR of HBV-DNA in sera and FQ-PCR in cultured supernatants of PBMC from hepatitis B types.They were not obviously related with the levels of ALT and TBiL.Conclusions The IL-12 and γ IFN secretion increased when the HBV-DNA of PBMC from carriers of HBeAg were at high-reproducing state.The HBV-DNA positive rate in cultured supernatants of PBMC of grave hepatitis detected by FQ-PCR was higher than that in sera by qualitative PCR.