Charbit A, Molla A, Ronco J, Clément J M, Favier V, Bahraoui E M, Montagnier L, Leguern A, Hofnung M
Unité de Programmation Moléculaire et Toxicologie Génétique (CNRS UA271, INSERM U163), Paris, France.
AIDS. 1990 Jun;4(6):545-51. doi: 10.1097/00002030-199006000-00008.
We expressed peptides from the HIV-1 envelope protein at the surface of Escherichia coli by genetic insertions into an exposed loop of the outer membrane protein LamB. Recombinant bacteria expressing eight peptides from gp110 (pep1-pep8), conserved between HIV-1 and HIV-2, were used as live immunogens in rabbits by the intravenous route. The eight constructions elicited anti-LamB antibodies, showing that the hybrid proteins were immunogenic. One of them, LamB-pep8, gave rise to antibodies able to react with gp160 and to neutralize HIV-1 in vitro. We also show that this type of recombinant E. coli can provide a convenient reagent to monitor and characterize specific antibodies. Recombinant clones were used to test sera of seropositive individuals, as well as to narrow down the monoclonal antibody 110-1 recognition site to a cluster of eight residues at the carboxy-terminal end of gp110.
通过基因插入外膜蛋白LamB的一个暴露环，我们在大肠杆菌表面表达了来自HIV-1包膜蛋白的肽段。表达来自gp110的八个肽段（pep1-pep8）的重组细菌，这些肽段在HIV-1和HIV-2之间保守，通过静脉途径用作家兔的活免疫原。这八个构建体引发了抗LamB抗体，表明杂合蛋白具有免疫原性。其中之一，LamB-pep8，产生了能够与gp160反应并在体外中和HIV-1的抗体。我们还表明，这种类型的重组大肠杆菌可以提供一种方便的试剂来监测和表征特异性抗体。重组克隆用于检测血清反应阳性个体的血清，以及将单克隆抗体110-1的识别位点缩小到gp110羧基末端的八个残基簇。
