Poss M L, Quackenbush S L, Mullins J I, Hoover E A
Department of Pathology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523.
J Virol. 1990 Sep;64(9):4338-45. doi: 10.1128/JVI.64.9.4338-4345.1990.
FeLV-FAIDS, an immunodeficiency-inducing isolate of feline leukemia virus, is composed of a pathogenic but replication-defective genome (molecular clone 61C) and a replication-competent but non-immunodeficiency-inducing variant genome (molecular clone 61E). The chimeric virus EECC, composed of the 5' gag-pol of 61E fused to the env-3' LTR of 61C, also induces immunodeficiency. The 61C (or EECC) gp80 can be distinguished from that of 61E on the basis of antigenic recognition, size, and rate of posttranslational processing. We found that the nascent precursor polypeptides of the two viruses were the same size; however, the 61E gp80 rapidly shifted to a smaller size and was subsequently cleaved to gp70, whereas EECC gp80 maintained its nascent size and was cleaved to gp70 only after a prolonged time. Endo-beta-N-acetyl glucosaminidase H and N-glycanase digestions of newly formed glycoproteins resulted in a similar banding pattern for both viruses, indicating that both contained the same number of oligosaccharide side chains and that all of these were high mannose sugars. The metabolic inhibitors of glycosylation, castanospermine or N-methyldeoxynojirimycin, prevented both the rapid trimming of 61E gp80 and its cleavage to gp70. Treatment with mannosidase inhibitors, however, did not affect 61E gp80 processing or size, suggesting that retention of glucose residues on EECC was responsible for these distinguishing properties of the glycoprotein. The pathological consequence of aberrant viral glycoprotein processing was evaluated in feline 3201 T lymphocytes, which are infectable by both 61E and EECC but are killed only by EECC. As in fibroblasts, the EECC glycoprotein produced in lymphocytes was larger, antigenically distinct, and processed more slowly than was the glycoprotein of 61E. Castanospermine treatment of 61E-infected 3201 T cells, however, not only abrogated the antigenic differences between the 61E and EECC glycoproteins but also resulted in a cytopathic effect. Our results suggest that (i) intracellular accumulation of EECC envelope glycoprotein may occur consequent to retention of glucose residues on carbohydrate side chains and (ii) a strong correlation exists between delayed glycoprotein processing and cytopathicity in FeLV-FAIDS-infected T lymphocytes.
猫白血病病毒致免疫缺陷型分离株(FeLV - FAIDS)由一个致病但复制缺陷的基因组(分子克隆61C)和一个复制能力完整但不致免疫缺陷的变异基因组(分子克隆61E)组成。嵌合病毒EECC由61E的5'gag - pol与61C的env - 3'LTR融合而成,也会诱发免疫缺陷。基于抗原识别、大小和翻译后加工速率,61C(或EECC)的gp80可与61E的gp80区分开来。我们发现这两种病毒的新生前体多肽大小相同;然而,61E的gp80迅速转变为较小的大小,随后裂解为gp70,而EECC的gp80保持其新生大小,仅在较长时间后才裂解为gp70。对新形成的糖蛋白进行内切β - N - 乙酰葡糖胺酶H和N - 聚糖酶消化,两种病毒产生了相似的条带模式,表明两者含有相同数量的寡糖侧链,且所有这些侧链都是高甘露糖糖。糖基化代谢抑制剂栗精胺或N - 甲基脱氧野尻霉素可阻止61E的gp80快速修剪及其裂解为gp70。然而,用甘露糖苷酶抑制剂处理并不影响61E的gp80加工或大小,这表明EECC上葡萄糖残基的保留是该糖蛋白这些独特性质的原因。在猫3201 T淋巴细胞中评估了异常病毒糖蛋白加工的病理后果,61E和EECC均可感染该细胞,但只有EECC会将其杀死。与成纤维细胞一样,淋巴细胞中产生的EECC糖蛋白比61E的糖蛋白更大,抗原性不同,加工也更慢。然而,用栗精胺处理感染61E的3201 T细胞,不仅消除了61E和EECC糖蛋白之间的抗原差异,还导致了细胞病变效应。我们的结果表明:(i)由于碳水化合物侧链上葡萄糖残基的保留,EECC包膜糖蛋白可能会在细胞内积累;(ii)在FeLV - FAIDS感染的T淋巴细胞中,糖蛋白加工延迟与细胞病变之间存在很强的相关性。