Cruz L J, Quintana D, Iglesias E, Garcia Y, Huerta V, Garay H E, Duarte C, Reyes O
División de Quimíca-Física, Centro de Ingeniería Genética y Biotecnología, Cubanacán, Habana, Cuba.
J Pept Sci. 2000 May;6(5):217-24. doi: 10.1002/(SICI)1099-1387(200005)6:5<217::AID-PSC242>3.0.CO;2-U.
The multiple antigenic peptide system (MAP) has been proposed as a novel and valuable approach for eliciting antibodies for peptides and developing synthetic vaccines. Multi-epitope polypeptides (MEP) have also been developed as an alternative to the recombinant approach for vaccines. The V3 loop from the HIV type 1 (HIV-1) external glycoprotein (gp120) contains the principal neutralization domain (PND). Antibodies against this region neutralize HIV-1 in vitro and in vivo. In this work, a novel presentation of di-epitope MAP was synthesized. A monomeric MAP carrying two identical JY1 V3 sequences as B-cell epitopes and the 830-843 region of tetanus toxoid as a T-helper cell epitope was synthesized. This basic structure was covalently linked to produce a four-JY1-branched homodimer (JY1-MAP4). Additionally, six different monomeric MAPs, bearing four copies of V3 from isolates LR150, JY1, RF, MN, BRVA and IIIB, were synthesized. These monomers were conveniently linked among themselves to produce homodimeric and heterodimeric MAPs of eight V3 branches (V3-MAP8). JY1-MAP8 elicited higher antibody titers in Balb/c mice than JY1-MAP4. The immunogenicity of two different, hexavalent V3-MAP8 mixtures and the MEP TAB9, which tandems the same six V3 sequences in a single molecule, were compared. The antibody response against the mixtures of the heterodimeric MAP showed a wider recognition pattern of the V3 region, while the homodimeric cocktail showed an intermediate pattern. Antibodies elicited by TAB9 recognized only the JY1, LR150 peptides. These results emphasize the influence of V3 epitope presentation upon the characteristics of the antibody response generated.
多抗原肽系统（MAP）已被提议作为一种用于引发针对肽的抗体和开发合成疫苗的新颖且有价值的方法。多表位多肽（MEP）也已被开发出来，作为疫苗重组方法的替代方案。来自1型人类免疫缺陷病毒（HIV-1）外膜糖蛋白（gp120）的V3环包含主要中和结构域（PND）。针对该区域的抗体在体外和体内均可中和HIV-1。在这项工作中，合成了一种新型的双表位MAP。合成了一种单体MAP，其携带两个相同的JY1 V3序列作为B细胞表位，以及破伤风类毒素的830 - 843区域作为T辅助细胞表位。这种基本结构通过共价连接产生了一种四分支的JY1同型二聚体（JY1-MAP4）。此外，还合成了六种不同的单体MAP，它们分别带有来自分离株LR150、JY1、RF、MN、BRVA和IIIB的四个V3拷贝。这些单体可方便地相互连接，产生具有八个V3分支的同型二聚体和异型二聚体MAP（V3-MAP8）。JY1-MAP8在Balb/c小鼠中引发的抗体滴度高于JY1-MAP4。比较了两种不同的六价V3-MAP8混合物和MEP TAB9（其在单个分子中串联相同的六个V3序列）的免疫原性。针对异型二聚体MAP混合物的抗体反应显示出对V3区域更广泛的识别模式，而同型二聚体混合物则显示出中等模式。由TAB9引发的抗体仅识别JY1、LR150肽。这些结果强调了V3表位呈现对所产生抗体反应特征的影响。
