Schiappacassi M, Buratti E, D'Agaro P, Ciani L, Scodeller E S, Tisminetzky S G, Baralle F E
International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy.
J Virol Methods. 1997 Jan;63(1-2):121-7. doi: 10.1016/s0166-0934(96)02120-9.
We have reported recently a new epitope presenting system based on the Flock House Virus (FHV) capsid protein. The HIV-1 V3 loop core sequence IGPGRAF was inserted in different sites of this carrier molecule. Immunoreactivity experiments and molecular modelling consistently showed that the most reactive recombinant protein displayed the IGPGRAF sequence in a conformation which is most similar to that of a V3 loop reference structure. The same insertion site was then used to display the V3 loop apex sequences of six different HIV-1 isolates. Sera from 32 HIV-1 infected patients were examined for their reactivity to our chimeric proteins and the results were compared with those obtained using synthetic V3 loop peptides. The data obtained were confirmed by nested PCR amplification and direct sequencing of the patient's V3 loops. The results showed that the V3 loop serotyping using the FHV hybrid proteins, was more specific than that obtained using synthetic peptides. This system will therefore be a useful tool for the correct evaluation of the immune response against different V3 loop core sequences.
我们最近报道了一种基于鸡瘟病毒（FHV）衣壳蛋白的新型表位呈递系统。HIV-1 V3环核心序列IGPGRAF被插入到该载体分子的不同位点。免疫反应性实验和分子建模一致表明，反应性最强的重组蛋白所展示的IGPGRAF序列的构象与V3环参考结构的构象最为相似。然后使用相同的插入位点展示六种不同HIV-1分离株的V3环顶端序列。检测了32名HIV-1感染患者血清对我们嵌合蛋白的反应性，并将结果与使用合成V3环肽获得的结果进行比较。通过巢式PCR扩增和对患者V3环的直接测序证实了所获得的数据。结果表明，使用FHV杂交蛋白进行V3环血清分型比使用合成肽更具特异性。因此，该系统将成为正确评估针对不同V3环核心序列的免疫反应的有用工具。
