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一种含有对人II型嗜T细胞病毒具有特异性表位的重组抗原的克隆与分析

Cloning and analysis of a recombinant antigen containing an epitope specific for human T-cell lymphotropic virus type II.

作者信息

Hadlock K G, Lipka J J, Chow T P, Foung S K, Reyes G R

机构信息

Department of Molecular Virology, Genelabs, Inc, Redwood City, CA 94063.

出版信息

Blood. 1992 May 15;79(10):2789-96.

PMID:1375112
Abstract

An immunodominant HTLV-I-specific epitope in the HTLV-I envelope glycoprotein (GP) 46 has been described. To determine if the analogous region of HTLV-II contains a similarly immunogenic and specific epitope, the polymerase chain reaction (PCR) was used to amplify HTLV-II DNA fragments encoding various portions of the putative epitope. The synthesized DNAs were cloned into lambda-phage gt11 and screened for production of immunoreactive fusion protein using sera from HTLV-II- or HTLV-I-infected individuals. Antisera from HTLV-II-infected individuals identified three of four recombinant clones when tested in a plaque immunoassay. Fusion protein from one of the clones, GH2-K15, was purified and analyzed by Western blot against a panel of HTLV-I and HTLV-II antisera. Twenty-one of 22 HTLV-II-infected sera were reactive with the GH2-K15 epitope. Sera from HTLV-I-infected and HTLV-I-uninfected individuals did not cross-react with GH2-K15. Western blot analysis of recombinant proteins encoding portions of the HTLV-II sequences in the Gh2-K15 antigen localized the HTLV-II-specific epitope to a 17-amino acid sequence. Recombinant antigens containing this epitope should be useful for type-specific serologic diagnosis of HTLV-II infection.

摘要

已发现人类嗜T淋巴细胞病毒I型(HTLV-I)包膜糖蛋白(GP)46中有一个免疫显性的HTLV-I特异性表位。为确定HTLV-II的类似区域是否含有同样具有免疫原性和特异性的表位,采用聚合酶链反应(PCR)扩增编码假定表位各部分的HTLV-II DNA片段。将合成的DNA克隆到λ噬菌体gt11中,并用HTLV-II或HTLV-I感染者的血清筛选免疫反应性融合蛋白的产生。在噬菌斑免疫测定中,HTLV-II感染者的抗血清识别出四个重组克隆中的三个。对其中一个克隆GH2-K15的融合蛋白进行纯化,并针对一组HTLV-I和HTLV-II抗血清进行蛋白质印迹分析。22份HTLV-II感染者血清中有21份与GH2-K15表位发生反应。HTLV-I感染者和未感染HTLV-I个体的血清不与GH2-K15发生交叉反应。对编码Gh2-K15抗原中HTLV-II序列部分的重组蛋白进行蛋白质印迹分析,将HTLV-II特异性表位定位到一个17个氨基酸的序列。含有该表位的重组抗原应可用于HTLV-II感染的型特异性血清学诊断。

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