In vitro assay for neutralizing antibody to hepatitis C virus:: Evidence for broadly conserved neutralization epitopes
In vitro assay for neutralizing antibody to hepatitis C virus:: Evidence for broadly conserved neutralization epitopes
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DOI：
10.1073/pnas.2335981100
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发表时间：
2003-11-25
影响因子：
11.1
通讯作者：
Purcell, RH
中科院分区：
文献类型：
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作者：
Bartosch, B;Bukh, J;Purcell, RH
我们对丙型肝炎病毒（HCV）体液免疫应答的了解有限，因为该病毒只能在人类和黑猩猩中进行研究，而且先前描述的中和试验不够稳健或操作复杂。然而，流行病学和实验室研究表明，针对HCV的中和抗体在预防感染方面可能很重要。我们最近描述了一种基于中和表面带有HCV包膜糖蛋白的假型鼠逆转录病毒构建体的中和试验。我们将该试验应用于来自HCV感染患者和黑猩猩的特征明确的临床样本，证实了针对HCV的中和抗体的存在，并验证了大多数先前报道的病毒中和情况。我们在病情缓解的感染中未发现中和性抗 - HCV，但在慢性感染中发现了相对较高滴度（>1:320）的此类抗体。中和抗体不仅针对E2包膜蛋白高变区的表位，还针对病毒包膜其他部位的一个或多个表位。中和抗体具有广泛的反应性，能够中和携带两种不同亚基因型（1a和1b）包膜糖蛋白的假型颗粒。检测中和性抗 - HCV的能力应能评估通过抗体介导的针对丙型肝炎的被动和主动免疫预防成功的前景。
Our understanding of the humoral immune response to hepatitis C virus (HCV) is limited because the virus can be studied only in humans and chimpanzees and because previously described neutralization assays have not been robust or simple to perform. Nevertheless, epidemiologic and laboratory studies suggested that neutralizing Ab to HCV might be important in preventing infection. We have recently described a neutralization assay based on the neutralization of pseudotyped murine retrovirus constructs bearing HCV envelope glycoproteins on their surface. We have applied the assay to well characterized clinical samples from HCV-infected patients and chimpanzees, confirmed the existence of neutralizing Ab to HCV, and validated most previously reported neutralizations of the virus. We did not find neutralizing anti-HCV in resolving infections but did find relatively high titers (>1:320) of such Ab in chronic infections. Neutralizing Ab was directed not only to epitope(s) in the hypervariable region of the E2 envelope protein but also to one or more epitopes elsewhere in the envelope of the virus. Neutralizing Ab was broadly reactive and could neutralize pseudotype particles bearing the envelope glycoproteins of two different subgenotypes (1a and 1b). The ability to assay neutralizing anti-HCV should permit an assessment of the prospects for successful Ab-mediated passive and active immunoprophylaxis against hepatitis C.