Boucher F D, Yasukawa L L, Kerns K, Kastelein M, Arvin A M, Prober C G
Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA.
Clin Diagn Virol. 1993 Mar;1(1):29-38. doi: 10.1016/0928-0197(93)90031-y.
The gene (US4) coding for herpes simplex virus type 2 (HSV-2) glycoprotein G (gG-2) was cloned and constitutively expressed in Chinese hamster ovary (CHO) cells. The expression vector containing the dihydrofolate reductase (dhfr) gene, and the HSV-2 US4 gene under the control of the Simian virus 40 early promoter (SV40 EP), was transfected into dhfr-deficient CHO cells. The transfected cells were selected and amplified using methotrexate (MTX). To demonstrate that the gG-2 produced in these transformed cells had antigenic determinants in common with the native glycoprotein, CHO cells expressing gG-2 were used in an immunofluorescent assay (IFA) for the detection of HSV-2 type-specific antibodies in human serum samples. Seven of eight serum samples from adults with prior episodes of culture proven HSV-2 infections were found to be positive by the IFA method whereas none of seven serum samples from young children with culture documented HSV-1 infections were positive by IFA. Thus the recombinant CHO : gG-2 cells have diagnostic utility in an HSV-2 specific serologic assay.
编码单纯疱疹病毒2型（HSV - 2）糖蛋白G（gG - 2）的基因（US4）被克隆，并在中国仓鼠卵巢（CHO）细胞中组成型表达。将含有二氢叶酸还原酶（dhfr）基因以及在猴病毒40早期启动子（SV40 EP）控制下的HSV - 2 US4基因的表达载体转染到缺乏dhfr的CHO细胞中。使用甲氨蝶呤（MTX）对转染的细胞进行筛选和扩增。为了证明在这些转化细胞中产生的gG - 2与天然糖蛋白具有共同的抗原决定簇，将表达gG - 2的CHO细胞用于免疫荧光测定（IFA），以检测人血清样本中的HSV - 2型特异性抗体。通过IFA方法发现，八份来自有培养证实的HSV - 2既往感染史的成人血清样本中有七份呈阳性，而七份来自有培养记录的HSV - 1感染的幼儿血清样本中没有一份通过IFA呈阳性。因此，重组CHO : gG - 2细胞在HSV - 2特异性血清学检测中具有诊断效用。
