Girerd Y, Cassé H, Duc Dodon M, Gazzolo L
Immuno-Virologie Moléculaire et Cellulaire, UMR30, Centre National de la Recherche Scientifique, Université Claude Bernard Lyon I, France.
J Gen Virol. 1995 Apr;76 ( Pt 4):1021-4. doi: 10.1099/0022-1317-76-4-1021.
The envelope (env) gene of human T cell leukaemia virus type I (HTLV-I) was inserted into an expression vector, referred to as phMTenv, under the transcriptional control of the human metallothionein IIa gene promoter (hMT-IIa). When this vector was transiently transfected in HeLa cells treated with hMT-IIa inducers, formation of multinucleated cells was observed, indicating the expression of functional surface and transmembrane glycoproteins. Of several HeLa cell clones transfected with phMTenv together with a plasmid carrying the neomycin resistance gene and isolated after selection in G418-containing medium, env mRNA was detected in only two, in the presence of hMT-IIa inducers. Viral glycoproteins were found to be weakly expressed as detected in immunoprecipitation assays of 125I-surface-labelled cells. These env-transfected HeLa cell clones, although unable to form syncytia when cocultivated with untransfected control HeLa cells, retained the capacity to fuse with HTLV-I-producing C91PL T cells. However, a significant decrease in their fusogenic ability was observed, after treatment with hMT-IIa inducers. Under identical experimental conditions, control HeLa cell clones stably transformed with the same plasmid, but lacking the env gene, were still able to fuse with C91PL cells. These observations suggest that a post-transcriptional step in HTLV-I env expression is impaired, probably leading to the establishment of superinfection interference.
将人类I型T细胞白血病病毒(HTLV-I)的包膜(env)基因插入到一个表达载体中,该载体称为phMTenv,处于人金属硫蛋白IIa基因启动子(hMT-IIa)的转录控制之下。当用hMT-IIa诱导剂处理的HeLa细胞中瞬时转染该载体时,观察到多核细胞的形成,这表明功能性表面糖蛋白和跨膜糖蛋白得以表达。在用携带新霉素抗性基因的质粒与phMTenv一起转染并在含G418的培养基中筛选后分离得到的几个HeLa细胞克隆中,仅在两个克隆中检测到env mRNA,且是在存在hMT-IIa诱导剂的情况下。在对125I表面标记细胞的免疫沉淀试验中检测到病毒糖蛋白表达较弱。这些env转染的HeLa细胞克隆,虽然在与未转染的对照HeLa细胞共培养时不能形成多核巨细胞,但仍保留与产生HTLV-I的C91PL T细胞融合的能力。然而,在用hMT-IIa诱导剂处理后,观察到它们的融合能力显著下降。在相同的实验条件下,用相同质粒稳定转化但缺乏env基因的对照HeLa细胞克隆仍能与C91PL细胞融合。这些观察结果表明,HTLV-I env表达中的转录后步骤受损,可能导致建立超感染干扰。