Hunt J C, Golden A M, Lund J K, Gürtler L G, Zekeng L, Obiang J, Kaptué L, Hampl H, Vallari A, Devare S G
AIDS Research and Retrovirus Discovery, Abbott Laboratories, North Chicago, Illinois 60064, USA.
AIDS Res Hum Retroviruses. 1997 Aug 10;13(12):995-1005. doi: 10.1089/aid.1997.13.995.
Four sera from Equatorial Guinea (EG) suspected to contain antibody against HIV-1 group O-related viruses were identified on the basis of unusual and differential serologic reactivity in selected commercial assays and Western blot. Degenerate primers, designed from HIV-1 group O published sequences, were used to PCR amplify envelope (env) gene sequences from the suspect EG sera. A complete envelope gene sequence from each serum was determined from the overlapping env gene fragments. Analysis (PHYLIP package of programs) of Env amino acid sequences (translated from nucleotide sequences) indicated that the amino acid sequences obtained from EG sera clustered more closely with HIV Env sequences of group O compared to group M. The amino acid sequences at the octameric tip of the V3 loop were either RIGPLAWY (one isolate), RIGPMAWY (two isolates), or GLGPLAVY (one isolate). The V3 tip tetrameric sequence GPLA is represented only once in the 1995 HIV (Los Alamos) database, but was present in two of our group O-related EG samples. The gp41 immunodominant regions (IDR) protein sequences were identical for sequences from three of the sera, RLLALETLIQNQQLLNLWGCKGR(K)L(I)VCYTSVK(T)W, whereas sequence from the fourth serum contained three changes as noted in parentheses. IDR sequences derived from EG sera were unique compared to those reported for other HIV-1 group O isolate ANT70, VAU, or MVP5180. Antibody in each EG serum directed against the IDR could be detected using synthetic peptides comprising sequences from the ANT70 or MVP5180 IDRs, but were most reactive against the sequences derived from the samples themselves. Little or no serologic reactivity was detected when EG sera were reacted against peptides comprising the IDR of HIV-1 group M (subtype B consensus) or HIV-2 (consensus).
在选定的商业检测和蛋白质印迹中,基于异常和不同的血清学反应性,鉴定出4份来自赤道几内亚(EG)的疑似含有抗HIV-1 O组相关病毒抗体的血清。根据已公布的HIV-1 O组序列设计简并引物,用于从疑似EG血清中PCR扩增包膜(env)基因序列。从重叠的env基因片段中确定每份血清的完整包膜基因序列。对Env氨基酸序列(从核苷酸序列翻译而来)的分析(使用PHYLIP程序包)表明,与M组相比,从EG血清中获得的氨基酸序列与O组HIV Env序列聚类更紧密。V3环八聚体末端的氨基酸序列为RIGPLAWY(1个分离株)、RIGPMAWY(2个分离株)或GLGPLAVY(1个分离株)。V3末端四聚体序列GPLA在1995年HIV(洛斯阿拉莫斯)数据库中仅出现一次,但在我们的两个O组相关EG样本中存在。三份血清的序列中gp41免疫显性区域(IDR)蛋白序列相同,为RLLALETLIQNQQLLNLWGCKGR(K)L(I)VCYTSVK(T)W,而第四份血清的序列包含括号中所示的三个变化。与其他HIV-1 O组分离株ANT70、VAU或MVP5180报道的IDR序列相比,EG血清来源的IDR序列是独特的。使用包含ANT70或MVP5180 IDR序列的合成肽可以检测到每份EG血清中针对IDR的抗体,但对来自样本本身的序列反应最强。当EG血清与包含HIV-1 M组(B亚型共识)或HIV-2(共识)IDR的肽反应时,几乎检测不到或没有血清学反应。