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目的观察慢性乙型肝炎(CHB)患者拉米夫定抗病毒治疗前后外周血HBV特异性T辅助细胞频率的变化及与血清HBVDNA的关系。方法用重组人HBcAg作为刺激抗原,采用酶联免疫斑点法(ELISPOT)检测25例CHB轻、中度患者外周血抗病毒治疗前后分泌IFNγ的HBV特异性T辅助细胞的频率。用即时定量荧光PCR仪定量检测患者血清HBVDNA水平。结果CHB轻、中度患者治疗前外周血HBV特异性T辅助细胞频率均值为(47.30±25.50)SFCs/1×106PBMC,拉米夫定抗病毒治疗3个月后显著降低(23.10±18.45SFCs/1×106PBMC,P<0.05)。动态观察的8例CHB轻、中度患者抗病毒治疗前HBV特异性T辅助细胞频率均高于治疗后;6例患者抗病毒治疗后血清HBVDNA水平显著降低,2例患者血清HBVDNA水平没有下降,血清HBVDNA水平下降的患者(除1例外)治疗前HBV特异性T辅助细胞频率高于血清HBVDNA水平没有下降的患者。结论炎症期CHB轻、中度患者HBV特异性T辅助细胞反应高于炎症缓解期。
Objective To investigate the changes of HBV-specific T-helper cells in peripheral blood before and after lamivudine treatment in patients with chronic hepatitis B (CHB) and its relationship with serum HBV DNA. Methods Recombinant human HBcAg was used as stimulating antigen and ELISPOT was used to detect the frequency of HBV-specific T helper cells secreting IFNγ before and after antiviral treatment in 25 mild and moderate CHB patients. Quantitative detection of serum HBVDNA levels using real-time quantitative PCR machine. Results The average frequency of HBV-specific T helper cells in pre-treatment CHB patients was (47.30 ± 25.50) SFCs / 1 × 106 PBMCs, and the lamivudine antiviral therapy significantly decreased after 3 months (23.10 ± 18.45 SFCs / 1 × 106 PBMC, P <0.05). The frequency of HBV-specific T-helper cells in 8 CHB patients before and after antiviral treatment was significantly higher than that of the control group. The serum HBV DNA levels in 6 patients were significantly decreased after antiviral treatment, while the serum HBV DNA levels did not decrease in 2 patients. Serum HBVDNA levels decreased (except 1) before treatment, HBV-specific T helper cell frequency higher than serum HBVDNA levels did not decline in patients. Conclusions HBV-specific T-helper responses in mild and moderate CHB patients during inflammation are higher than those in remission.