Therrien D, St-Pierre Y, Dea S
Centre de Microbiologie et Biotechnologie, INRS-Institut Armand-Frappier, Université du Québec, Laval, Canada.
Arch Virol. 2000;145(6):1099-116. doi: 10.1007/s007050070112.
In its natural host, porcine reproductive and respiratory syndrome virus (PRRSV) has been reported to have a restricted tropism for cells of the monocyte/macrophage lineage. To date, cloned monkey kidney cell lines, such as MARC-145 and CL2621 cells which have been established from MA-104 cells, are the only non-porcine cells known to support PRRSV replication. In the present study, a binding assay was set up to follow by flow cytometry the attachment of PRRSV on the surface of porcine and non-porcine cells. PRRSV was found to be able to bind permissive cells like porcine alveolar macrophages and MARC-145. Further binding assays with porcine peripheral blood leukocytes showed that only monocytes can attach the virus. By their lack of binding factor, lymphocytes appeared to be refractory to PRRSV infection. Pre-incubation of MARC-145 cells with chymotrypsin and pronase E, but not neuraminidase, blocked their binding activity for PRRSV. The binding activity of the protease-treated cells was regenerated 8 hours after treatment, but cells remained unable to bind PRRSV if maintained in the presence of cycloheximide, thus confirming the proteinic nature of the specific binding factor(s). Experiments conduced with cells that have been previously characterized as non-permissive to PRRSV infection showed that many of them were able to bind the virus. Data obtained suggest that interaction of PRRSV with a specific binding factor on the surface of some cells is not sufficient to lead to a productive infection, and that a second putative receptor or other phenomena are probably required to pursue later events.
据报道,猪繁殖与呼吸综合征病毒(PRRSV)在其自然宿主中对单核细胞/巨噬细胞系的细胞具有有限的嗜性。迄今为止,克隆的猴肾细胞系,如从MA - 104细胞建立的MARC - 145和CL2621细胞,是已知的仅有的能支持PRRSV复制的非猪细胞。在本研究中,建立了一种结合试验,通过流式细胞术追踪PRRSV在猪和非猪细胞表面的附着情况。发现PRRSV能够结合允许性细胞,如猪肺泡巨噬细胞和MARC - 145。对猪外周血白细胞进行的进一步结合试验表明,只有单核细胞能附着该病毒。由于缺乏结合因子,淋巴细胞似乎对PRRSV感染具有抗性。用胰凝乳蛋白酶和链霉蛋白酶E而非神经氨酸酶对MARC - 145细胞进行预孵育,可阻断它们对PRRSV的结合活性。蛋白酶处理后的细胞在处理8小时后恢复结合活性,但如果在放线菌酮存在的情况下培养,细胞仍无法结合PRRSV,从而证实了特异性结合因子的蛋白质性质。对先前已被鉴定为对PRRSV感染不允许的细胞进行的实验表明,其中许多细胞能够结合该病毒。获得的数据表明,PRRSV与某些细胞表面的特异性结合因子相互作用不足以导致有效感染,可能还需要第二个假定的受体或其他现象来推动后续事件。