Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Av. Universidad 2001, Col. Chamilpa, Cuernavaca, Morelos CP 62210, Mexico.
Virus Res. 2010 Feb;147(2):231-41. doi: 10.1016/j.virusres.2009.11.005. Epub 2009 Nov 20.
In this work we evaluated the ability of rotavirus strains with different receptor requirements to infect the apical and basolateral surfaces of polarized MDCKII cells. We used neuraminidase (NA)-sensitive (RRV and TFR-1) and neuraminidase-resistant (Wa and UK) viruses that differ in their use of integrins. Regardless of their receptor requirements, all virus strains tested were found to efficiently infect cells from both membrane surface domains, with preference for the basolateral domain, since: (i) disruption of tight junctions of polarized cell monolayers by calcium chelation led to a reversible increase of rotavirus infectivity, (ii) the viruses infected preferentially the cells located at the borders of microcolonies of polarized cells, and (iii) in cells grown on a permeable support all four virus strains were able to start the infection by either plasma membrane domain. Preferential infection (5-11-fold more efficiently) of the basolateral surface correlated with the neuraminidase resistance of the virus strains, but not with their requirement for integrins, which in MDCKII cells seem to be used by all four viruses. The infection of both cell surface domains by RRV was found to depend on the presence of terminal sialic acids, since its infectivity was reduced by neuraminidase treatment of the cells and it was also blocked by incubation of the virus with glycophorin A. The efficient infection through the basolateral membrane surface of polarized cells might be relevant for the pathogenesis of rotavirus, especially given the recent reports of antigenemia and extraintestinal spread of the virus in children and animal models.
在这项工作中，我们评估了具有不同受体需求的轮状病毒株感染极化 MDCKII 细胞顶侧和基底外侧表面的能力。我们使用了神经氨酸酶（NA）敏感型（RRV 和 TFR-1）和神经氨酸酶抗性型（Wa 和 UK）病毒，它们在整合素的使用上存在差异。无论其受体要求如何，所有测试的病毒株都被发现能够有效地感染来自两个膜表面区域的细胞，并且偏爱基底外侧区域，因为：（i）通过钙螯合破坏极化细胞单层的紧密连接会导致轮状病毒感染力的可逆增加，（ii）病毒优先感染位于极化细胞微菌落边缘的细胞，（iii）在可渗透支持物上生长的细胞中，所有四种病毒株都能够通过质膜域之一开始感染。基底外侧表面的优先感染（效率提高 5-11 倍）与病毒株的神经氨酸酶抗性相关，但与它们对整合素的需求无关，在 MDCKII 细胞中，这四种病毒似乎都使用整合素。RRV 对两个细胞表面区域的感染被发现依赖于末端唾液酸的存在，因为其感染性被细胞的神经氨酸酶处理降低，并且它也被病毒与糖蛋白 A 的孵育所阻断。轮状病毒通过极化细胞的基底外侧膜表面的有效感染可能与轮状病毒的发病机制有关，特别是鉴于最近有报道称抗原血症和病毒在儿童和动物模型中的肠道外传播。
