Silva R F, Baluda M A
J Virol. 1980 Sep;35(3):766-74. doi: 10.1128/JVI.35.3.766-774.1980.
We have analyzed the avian myeloblastosis virus proteins in two types of leukemic myeloblasts: established myeloblastic cell lines (DU 1765 and DU 11157) and leukemic myeloblasts obtained from the peripheral blood of a leukemic C/E Spafas chicken (no. 21957). Using monospecific antisera for immunoprecipitation and polyacrylamide gel electrophoresis, we have detected gag gene-related proteins in the myeloblasts. The DU 1765 and DU 11157 cells contained a p100 protein which possessed antigenic determinants of the viral proteins p27, p19, p15, and p12. The p100 was not found in leukemic myeloblasts from Spafas chickens, and pulse-chase experiments showed that the p100 was not a precursor for the viral proteins. However, the p100 is present in uninfected line 15 chicken embryos. A pr76-like protein was identified in DU 1765 cells but migrated slightly further into gels than the pr76 of Spafas-derived leukemic myeloblasts. The Spafas-derived myeloblasts produced a pr60, whereas the DU 1765 cells contained instead a related protein of 62,000 daltons. Using anti-avian myeloblastosis virus gp85 sera, a glycoprotein of 120,000 daltons (gp120) was detected in all the tested leukemic myeloblasts. The gp120 was also present, in low amounts, in uninfected embyonic spleen and yolk sac cells. The anti-gp85 sera also precipitated a 27,000-dalton protein (h27) in these same cells. Both the gp120 and h27 could not be detected in either uninfected or myeloblastosis-associated virus-infected fibroblasts. Limited peptide hydrolysis revealed that h27 is different from the viral structural protein p27. In conclusion, monospecific antisera for gag and env gene products of avian myeloblastosis virus did not precipitate any unique or aberrant avian myeloblastosis virus protein from leukemic myeloblasts.
已建立的成髓细胞系(DU 1765和DU 11157)以及从一只患白血病的C/E Spafas鸡(编号21957)外周血中获取的白血病性成髓细胞。通过使用单特异性抗血清进行免疫沉淀和聚丙烯酰胺凝胶电泳,我们在成髓细胞中检测到了与gag基因相关的蛋白。DU 1765和DU 11157细胞含有一种p100蛋白,它具有病毒蛋白p27、p19、p15和p12的抗原决定簇。在Spafas鸡的白血病性成髓细胞中未发现p100,脉冲追踪实验表明p100不是病毒蛋白的前体。然而,p100存在于未感染的15号品系鸡胚中。在DU 1765细胞中鉴定出一种pr76样蛋白,但它在凝胶中的迁移距离比Spafas来源的白血病性成髓细胞中的pr76稍远。Spafas来源的成髓细胞产生一种pr60,而DU 1765细胞中含有一种相关的62000道尔顿的蛋白。使用抗禽成髓细胞瘤病毒gp85血清,在所有测试的白血病性成髓细胞中检测到一种120000道尔顿的糖蛋白(gp120)。gp120在未感染的胚胎脾脏和卵黄囊细胞中也少量存在。抗gp85血清在这些相同细胞中还沉淀出一种27000道尔顿的蛋白(h27)。在未感染或与成髓细胞瘤相关病毒感染的成纤维细胞中均未检测到gp120和h27。有限的肽水解显示h27与病毒结构蛋白p27不同。总之,针对禽成髓细胞瘤病毒gag和env基因产物的单特异性抗血清未从白血病性成髓细胞中沉淀出任何独特或异常的禽成髓细胞瘤病毒蛋白。