Kojima M, Osuga T, Tsuda F, Tanaka T, Okamoto H
Department of Internal Medicine, University of Tsukuba School of Medicine, Ibaraki-Ken, Japan.
Virology. 1994 Nov 1;204(2):665-72. doi: 10.1006/viro.1994.1582.
A human plasma (inoculum one) containing hepatitis C virus (HCV) was passaged through eight chimpanzees in three generations. Of 10 HCV clones propagated from it, 7 were different in respect to the hypervariable region of E2/NS1 glycoprotein and they were named clones A, B, C, etc. A chimpanzee received inoculum one, and clone A accounted for 7 of the 10 clones from his acute-phase plasma (inoculum two). Five chimpanzees received inoculum two, and clone A accounted for 5 to 9 of the 10 clones each from their preacute plasma, which were pooled to make inoculum three. Of 10 HCV clones from inoculum three, 4 were A, 5 were B/B', and the remaining 1 was C. Two chimpanzees received inoculum three or its CsCl fraction, and all 40 clones from their acute-phase plasma were A. Thus, clone A in inoculum one was selected by chimpanzees during three passages. HCV virions in the three inocula were separated into free and immunoglobulin-bound forms by sucrose density fractionation. HCV virions in inocula one and two were heterogeneous in the sequence of hypervariable region and predominantly free of immunoglobulins. By contrast, inoculum three contained both free virions of predominantly A and immunoglobulin-bound virions which were heterogeneous. Antibodies to the hypervariable region were determined by enzyme immunoassays with overlapping synthetic decapeptides. Antibodies to clone A were detected in one chimpanzee who received inoculum two, and those to clones B and C in two chimpanzees including him and in inoculum three. Antibodies were not detectable in inoculum one or two or in the other chimpanzees. These results indicate that antibodies to the hypervariable region of E2/NS1 glycoprotein would be protective and contribute toward the selective replication of HCV in chimpanzees.
一份含有丙型肝炎病毒（HCV）的人血浆（接种物一）在三代中通过了八只黑猩猩。从其中繁殖出的10个HCV克隆中，7个在E2/NS1糖蛋白的高变区方面有所不同，它们被命名为克隆A、B、C等。一只黑猩猩接受了接种物一，来自其急性期血浆（接种物二）的10个克隆中有7个是克隆A。五只黑猩猩接受了接种物二，来自它们急性前期血浆的10个克隆中，每个克隆的5至9个是克隆A，这些血浆被汇集起来制成接种物三。来自接种物三的10个HCV克隆中，4个是A，5个是B/B'，其余1个是C。两只黑猩猩接受了接种物三或其CsCl组分，来自它们急性期血浆的所有40个克隆都是A。因此，接种物一中的克隆A在三代传代过程中被黑猩猩选择。通过蔗糖密度分级分离，将三种接种物中的HCV病毒粒子分为游离和免疫球蛋白结合形式。接种物一和二中的HCV病毒粒子在高变区序列上是异质的，并且主要不含免疫球蛋白。相比之下，接种物三既含有主要为A的游离病毒粒子，也含有异质的免疫球蛋白结合病毒粒子。通过使用重叠合成十肽的酶免疫测定法测定高变区的抗体。在一只接受接种物二的黑猩猩中检测到了针对克隆A的抗体，在包括它在内的两只黑猩猩以及接种物三中检测到了针对克隆B和C的抗体。在接种物一或二中或其他黑猩猩中未检测到抗体。这些结果表明，针对E2/NS1糖蛋白高变区的抗体具有保护作用，并有助于HCV在黑猩猩中的选择性复制。
