Bongartz J P, Aubertin A M, Milhaud P G, Lebleu B
Institut de Génétique Moléculaire de Montpellier-UMR CNRS 9942, Montpellier, France.
Nucleic Acids Res. 1994 Nov 11;22(22):4681-8. doi: 10.1093/nar/22.22.4681.
Recently several groups reported a dramatic improvement of reporter gene transfection efficiency using a fusogenic peptide, derived from the Influenza hemagglutinin envelop protein. This peptide changes conformation at acidic pH and destabilizes the endosomal membranes thus resulting in an increased cytoplasmic gene delivery. We describe the use of a similar fusogenic peptide in order to improve the antiviral potency of antisense oligodeoxynucleotides (anti TAT) and oligophosphorothioates (S-dC28) on de novo HIV infected CEM-SS lymphocytes in serum-free medium. We observed as 5 to 10 fold improvement of the anti HIV activities of the phosphodiester antisense oligonucleotides after chemical coupling to the peptide in a one to one ratio by a disulfide or thioether bond. No toxicities were observed at the effective doses (0.1-1 microM). No sequence specificity was obtained and the fusogenic peptide possessed some antiviral activities on its own (IC50: 6 microM). A S-dC28-peptide disulfide linked conjugate and a streptavidin-peptide-biotinylated S-dC28 adduct showed similar activities as the free S-dC28 oligonucleotide (IC50: 0.1-1 nM). As expected, all the compounds were less potent in the presence of serum but the relative contribution of peptide coupling was maintained.
最近，有几个研究小组报道称，使用一种源自流感血凝素包膜蛋白的融合肽可显著提高报告基因的转染效率。这种肽在酸性pH值下会改变构象，使内体膜不稳定，从而增加细胞质中的基因传递。我们描述了使用一种类似的融合肽，以提高反义寡脱氧核苷酸（抗TAT）和寡磷硫酯（S-dC28）在无血清培养基中对新感染HIV的CEM-SS淋巴细胞的抗病毒效力。我们观察到，通过二硫键或硫醚键以1:1的比例将磷酸二酯反义寡核苷酸与该肽化学偶联后，其抗HIV活性提高了5至10倍。在有效剂量（0.1 - 1 microM）下未观察到毒性。未获得序列特异性，且融合肽自身具有一定的抗病毒活性（IC50：6 microM）。一种S-dC28 - 肽二硫键连接的共轭物和一种链霉亲和素 - 肽 - 生物素化的S-dC28加合物显示出与游离S-dC28寡核苷酸相似的活性（IC50：0.1 - 1 nM）。正如预期的那样，在有血清存在的情况下，所有化合物的效力都较低，但肽偶联的相对作用得以维持。
