Zhang Ting, Xin Ruolei, Zhang Qin, Bao Yihua, Wu Jianxin
Capital Institute of Pediatrics, Beijing 100020, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2002 Mar;16(1):44-7.
To characterize the intertype epitopes on human adenovirus (HAdV) hexon.
Based on computerized analysis on adenoviruses sequence of genomic alignment, antigenicity prediction and 3-D structure characteristics of hexon subunit, several peptides of hexon of adenoviruses were chosen to be synthesized or recombinant proteins of the hexon were expressed in E. coli by use of PGEX-5X. To identify the existence of intertype epitopes, the antisera raised with synthetic peptides or purified recombinant proteins were analyzed with Western blot and immunofluorescent assay.
The results of Western blot indicated that both peptide and recombinant antibodies showed specific reactivities with hexons of HADv-3, 4, 7 individually. Meanwhile, typical stain of immunofluorescence was found on HeLa cells infected with these HAdV by incubation with peptide as well as recombinant antibodies. Also, antibodies raised against peptide recognized the recombinant hexon protein in which a corresponding region of peptides was covered.
Most of the predicted intertype epitopes of HAdV hexon wer e exclusively found in synthetic peptides and recombinant proteins. These intertype epitopes showed to be continuous and sequential which could be employed for development of antibodies of diagnostic use.
鉴定人腺病毒（HAdV）六邻体上的型间表位。
基于对腺病毒基因组比对序列、抗原性预测及六邻体亚基三维结构特征的计算机分析，选择腺病毒六邻体的几种肽段进行合成，或利用PGEX - 5X在大肠杆菌中表达六邻体重组蛋白。为鉴定型间表位的存在，采用蛋白质印迹法和免疫荧光分析法对用合成肽段或纯化重组蛋白制备的抗血清进行分析。
蛋白质印迹结果表明，肽段抗体和重组抗体均分别与HAdV - 3、4、7的六邻体呈现特异性反应。同时，用肽段抗体及重组抗体孵育感染这些腺病毒的HeLa细胞后，发现有典型的免疫荧光染色。此外，针对肽段制备的抗体能够识别覆盖有相应肽段区域的重组六邻体蛋白。
HAdV六邻体的大多数预测型间表位仅存在于合成肽段和重组蛋白中。这些型间表位呈连续且有序排列，可用于开发诊断用抗体。
