Giacchino R, Facco F, Loy A, Timitilli A, Cirillo C, Navone C, Ciravegna B, Pisani N
Instituto Scientifico G. Gaslini, 1a Clinica Malattie Infettive Università di Genova.
Boll Ist Sieroter Milan. 1989;68(1):24-7.
The aim of the present work is to assess whether reactivation of viral replication occurred in children affected by chronic hepatitis undergoing long term immunosuppressive therapy. 123 serum samples belonging to 25 children were retrospectively evaluated for HBV-DNA and HBeAg. Sera were collected prior to and during the protocol treatment (steroids alone or with azathioprine). Presence of HBV-DNA was evaluated by means of molecular hybridization technique, using a radiolabelled probe of cloned HBV-DNA. Sera (100 1) were denatured and transferred into nylon membrane (spot) then prehybridized and hybridized. After washing in stringent conditions, the filter was exposed in autoradiographic cassette with Kodak film XOmat5. Positivity was semiquantitatively evaluated by blackening of the spot. Increase or appearance of HBV-DNA was observed in the sera from 24/25 pts. HBeAg became positive in 4/5 pts previously negative. Obtained data shows reactivation of viral replication during immunosuppressive therapy. Data are especially significant in those cases in which such activity has apparently ceased or not been detected; HBV may be considered as a latent virus.
本研究的目的是评估接受长期免疫抑制治疗的慢性肝炎患儿是否发生病毒复制再激活。对25名儿童的123份血清样本进行回顾性评估,检测乙肝病毒脱氧核糖核酸(HBV-DNA)和乙肝e抗原(HBeAg)。在方案治疗前及治疗期间(单独使用类固醇或联合硫唑嘌呤)采集血清。采用分子杂交技术,使用克隆的HBV-DNA放射性标记探针评估HBV-DNA的存在情况。将血清(100 μl)变性后转移至尼龙膜(斑点)上,然后进行预杂交和杂交。在严格条件下洗涤后,将滤膜与柯达XOmat5胶片一起放入放射自显影片盒中曝光。通过斑点变黑进行阳性结果的半定量评估。在24/25例患者的血清中观察到HBV-DNA增加或出现。4/5例先前HBeAg阴性的患者转为阳性。获得的数据表明免疫抑制治疗期间病毒复制再激活。在那些病毒复制活性明显停止或未被检测到的病例中,该数据尤其显著;HBV可被视为一种潜伏病毒。