Characterization of a novel human immunodeficiency virus type 1 neutralizable epitope within the immunodominant region of gp41.
Characterization of a novel human immunodeficiency virus type 1 neutralizable epitope within the immunodominant region of gp41.
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gp41 免疫显性区域内新型人类免疫缺陷病毒 1 型可中和表位的表征。
DOI：
10.1006/viro.2000.0269
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
Ugen,KE
中科院分区：
文献类型：
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作者：
Viveros,M;Dickey,C;Cotropia,JP;Gevorkian,G;Larralde,C;Broliden,K;Levi,M;Burgess,A;Cao,C;Weiner,DB;Agadjanyan,MG;Ugen,KE
以前，我们使用来自无症状人类免疫缺陷病毒1型（HIV-1）血清阳性供体的外周血单个核细胞产生人单克隆抗体。这些单克隆抗体之一（命名为克隆3，CL 3）识别跨膜包膜糖蛋白gp 41的免疫显性区域（簇I）内的10个氨基酸（GCSGKLICTT），并中和不同实验室分离株对靶细胞的感染。由于CL 3识别的表位具有两个半胱氨酸残基，可能在gp 41中产生二硫环，我们分析了我们的单克隆抗体与肽序列IWGCSGKLICTTAVP的环状和线性基序（残基600-614）的结合。CL 3抗体不与合成的环肽结合，但识别线性形式。然后产生针对线性和环状肽的两种多克隆兔血清。这两种抗血清都能与病毒糖蛋白gp 41和gp 160结合，但都不能中和HIV-1实验室分离株。使用一组丙氨酸取代的IWGCSGKLICTTAV肽，我们分析了多克隆抗血清和CL 3的结合。这些肽的多克隆抗血清的结合的配置文件是不同的CL 3相同的肽。这表明CL 3识别独特的可中和核心表位，其在用作兔中免疫原的环状或线性IWGCSGKLICTTAVP肽中均不具有免疫原性。
Previously, we generated human monoclonal antibodies using peripheral blood mononuclear cells from an asymptomatic human immunodeficiency virus type 1 (HIV-1)-seropositive donor. One of these monoclonal antibodies (designated clone 3, CL3) recognized 10 amino acids (GCSGKLICTT) within the immunodominant region (cluster I) of the transmembrane envelope glycoprotein gp41 and neutralized infection of target cells with different laboratory isolates. Because the epitope recognized by CL3 has two cysteine residues that could potentially produce a disulfide loop in gp41, we analyzed binding of our monoclonal antibody to the cyclic and linear motif of the peptide sequence IWGCSGKLICTTAVP (residues 600–614). The CL3 antibody did not bind to the synthetic cyclic peptide but did recognize the linear form. Two polyclonal rabbit sera against both the linear and cyclic peptides were then generated. Both antisera bound to viral glycoproteins gp41 and gp160, but neither sera neutralized HIV-1 laboratory isolates. Using a set of alanine-substituted IWGCSGKLICTTAV peptides, we analyzed binding of polyclonal antisera and CL3. The profile of binding of polyclonal antisera to these peptides was different from that of CL3 to the same peptides. This suggests that CL3 recognized a unique neutralizable core epitope, which was not immunogenic in either the cyclic or the linear IWGCSGKLICTTAVP peptides used as immunogens in the rabbits.