Chebloune Y, Rulka J, Cosset F L, Valsesia S, Ronfort C, Legras C, Drynda A, Kuzmak J, Nigon V M, Verdier G
Laboratoire de Biologie Cellulaire, Centre National de la Recherche Scientifique UMR106, Université Claude Bernard Lyon-I, Villeurbanne, France.
J Virol. 1991 Oct;65(10):5374-80. doi: 10.1128/JVI.65.10.5374-5380.1991.
The Rous-associated virus 1 env gene, which encodes the envelope gp85 and gp37 glycoproteins, was isolated and inserted in place of the v-erbB oncogene into an avian erythroblastosis virus-based vector, carrying the neo resistance gene substituted for the v-erbA oncogene, to generate the pNEA recombinant vector. A helper-free virus stock of the pNEA vector was produced on an avian transcomplementing cell line and used to infect primary chicken embryo fibroblasts (CEFs) or quail QT6 cells. These infected cells, selected with G418 (CEF/NEA and QT6/NEA, respectively) were found to be resistant to superinfections with subgroup A retroviruses. The CEF/NEA preparations were used as a cell-associated antigen to inoculate adult chickens by the intravenous route compared with direct inoculations of NEA recombinant helper-free virus used as a cell-free antigen. Chickens injected with the cell-associated antigen (CEF/NEA) exhibited an immune response demonstrated by induction of high titers of neutralizing antibodies and were found to be protected against tumor production after Rous sarcoma virus A challenge. Conversely, no immune response and no protection against Rous sarcoma virus A challenge were observed in chickens directly inoculated with cell-free NEA recombinant virus or in sham-inoculated chickens.
劳氏相关病毒1的env基因编码包膜糖蛋白gp85和gp37,该基因被分离出来,并取代v-erbB癌基因插入到一个基于禽成红细胞增多症病毒的载体中,该载体携带取代v-erbA癌基因的新霉素抗性基因,从而产生pNEA重组载体。pNEA载体的无辅助病毒毒株在禽转互补细胞系上产生,并用于感染原代鸡胚成纤维细胞(CEF)或鹌鹑QT6细胞。用G418筛选这些感染细胞(分别为CEF/NEA和QT6/NEA),发现它们对A亚群逆转录病毒的超感染具有抗性。与直接接种作为无细胞抗原的NEA重组无辅助病毒相比,将CEF/NEA制剂作为细胞相关抗原通过静脉途径接种成年鸡。注射细胞相关抗原(CEF/NEA)的鸡表现出免疫反应,表现为诱导产生高滴度的中和抗体,并且在受到劳氏肉瘤病毒A攻击后被发现能够抵抗肿瘤产生。相反,直接接种无细胞NEA重组病毒的鸡或假接种的鸡未观察到免疫反应,也未受到劳氏肉瘤病毒A攻击的保护。