Gómez C E, López-Campistrous A E, Duarte C A
División de Vacunas, Centro de Ingenieria Genética y Biotecnología, Cubanacán, Havana, Cuba.
J Virol Methods. 1998 Mar;71(1):7-16. doi: 10.1016/s0166-0934(97)00196-1.
The V3 loop of the gp120 envelope glycoprotein contains the principal neutralizing determinant of HIV-1. Many serological studies have been carried out to assess the reactivity of HIV-1 infected individuals against V3 loop synthetic peptides from different HIV-1 subtypes. V3 directed serology has also been used to demonstrate the association between ELISA reactivity and progression to AIDS in HIV patients, and to study the reactivity against the V3 region in sera from vaccinated animals and human volunteers. The advantage of the use of bovine serum albumin (BSA) conjugated V3 peptides over free V3 peptides for ELISA is described. 15 meric V3 peptides representing several HIV-1 isolates were synthesized, chemically coupled to BSA, and used to coat ELISA microplates. Conjugated peptides were compared with free peptides for the detection of anti V3 antibodies in the sera from rabbits immunized with V3 containing chimeric proteins and from HIV-1 infected individuals. No differences in reactivity against free or BSA-peptide were found for most rabbit sera, however human plasma recognized preferentially the BSA conjugated peptides. Although technically more complex, ELISA with BSA coupled V3 peptides is more sensitive and appropriate for serological studies of HIV-1 infected persons.
gp120包膜糖蛋白的V3环包含HIV-1的主要中和决定簇。已经开展了许多血清学研究,以评估HIV-1感染个体对来自不同HIV-1亚型的V3环合成肽的反应性。V3定向血清学也已用于证明HIV患者中ELISA反应性与进展为艾滋病之间的关联,并研究接种疫苗的动物和人类志愿者血清中针对V3区域的反应性。描述了在ELISA中使用牛血清白蛋白(BSA)偶联的V3肽优于游离V3肽的优势。合成了代表几种HIV-1分离株的15聚体V3肽,化学偶联到BSA上,并用于包被ELISA微孔板。将偶联肽与游离肽进行比较,以检测用含V3嵌合蛋白免疫的兔血清和HIV-1感染个体血清中的抗V3抗体。对于大多数兔血清,未发现对游离或BSA肽的反应性存在差异,然而人血浆优先识别BSA偶联肽。尽管技术上更复杂,但使用BSA偶联V3肽的ELISA对HIV-1感染者的血清学研究更敏感且更合适。