Barteling S J, Meloen R H, Wagenaar F, Gielkens A L
J Gen Virol. 1979 May;43(2):383-93. doi: 10.1099/0022-1317-43-2-383.
Strains of foot-and-mouth disease virus of types O1 and A10 were isolated which showed no significant loss of infectivity upon trypsinization. These 'trypsin-resistant' (TR) viruses were obtained by serial passage in BHK cells of virus that was trypsin-treated before inoculation of the cells. Three O1 isolates were cloned and studied further. Cell attachment of those TR O1 variants (OTR1) was not reduced by trypsinization, unlike that of parent virus. The polypeptide compositions of TR viruses as determined by SDS-polyacrylamide gel electrophoresis were identical with those of parent virus, with the exception of OTR1 which contained an additional polypeptide approx, 3000 daltons larger than VP1. After trypsinization, which normally cleaves VP1, the polypeptide composition of the three TR viruses (including OTR1) and of parent virus did not show any significant difference. In OTR1 both the additional virus protein and VP1 were cleaved into a P18 molecule and smaller fragments. The surface location of this additional polypeptide was confirmed by iodination experiments. It was shown by immunodiffusion experiments that only OTR1 differed from the parent virus. This antigenic change was present on the trypsin-sensitive part of the virus since trypsinized TR viruses (including OTR1) were antigenically identical to trypsinized parent virus. The electrophoretic mobilities of the three OTR viruses isolated, and of parent virus, differed somewhat before trypsinization. After trypsin-treatment, the mobilities of TR viruses were all increased to the same level; however, their rate of migration was lower than that of trypsin-treated parent virus. This lower mobility of trypsin-treated OTR viruses was the only difference which could be associated with retained infectivity.
分离出了O1型和A10型口蹄疫病毒毒株,这些毒株经胰蛋白酶处理后感染性无显著损失。这些“胰蛋白酶抗性”(TR)病毒是通过在BHK细胞中对接种前经胰蛋白酶处理的病毒进行连续传代获得的。对三个O1分离株进行了克隆并进一步研究。与亲本病毒不同,这些TR O1变体(OTR1)的细胞附着不受胰蛋白酶处理的影响。通过SDS-聚丙烯酰胺凝胶电泳测定的TR病毒的多肽组成与亲本病毒相同,但OTR1除外,它含有一种比VP1大约3000道尔顿的额外多肽。在通常切割VP1的胰蛋白酶处理后,三种TR病毒(包括OTR1)和亲本病毒的多肽组成没有显示出任何显著差异。在OTR1中,额外的病毒蛋白和VP1都被切割成一个P18分子和更小的片段。通过碘化实验证实了这种额外多肽的表面定位。免疫扩散实验表明只有OTR1与亲本病毒不同。这种抗原性变化存在于病毒的胰蛋白酶敏感部分,因为经胰蛋白酶处理的TR病毒(包括OTR1)与经胰蛋白酶处理的亲本病毒在抗原上是相同的。分离出的三种OTR病毒和亲本病毒在胰蛋白酶处理前的电泳迁移率有所不同。胰蛋白酶处理后,TR病毒的迁移率都增加到相同水平;然而,它们的迁移速度低于经胰蛋白酶处理的亲本病毒。经胰蛋白酶处理的OTR病毒这种较低的迁移率是与保留感染性相关的唯一差异。