新城疫病毒致敏红细胞的凝集-分离反应：神经氨酸酶再激活后释放的改变病毒和HN刺突的量、凝集特性及血清学

Agglutination-separation reactions of red blood cells sensitized with Newcastle disease virus: quantities, agglutination characteristics, and serology of altered virus and HN spikes released following neuraminidase reactivation.

作者信息

Graves I L

机构信息

Department of Molecular Microbiology and Immunology, School of Hygiene and Public Health, The Johns Hopkins University, Baltimore, Maryland 21205, USA.

出版信息

Vet Res. 2001 Sep-Oct;32(5):475-89. doi: 10.1051/vetres:2001139.

DOI:10.1051/vetres:2001139

PMID:11592617

Abstract

Red blood cells (RBC) become sensitized following the elution of strain 575 Newcastle disease virus (NDV). The neuraminidase (NA) in the haemagglutinin (HA)-sialic acid configuration is inactive. The HA on sensitized RBC agglutinates normal RBC. The sialic acid on normal RBC initiated reactivation of the NA-a newly described function. Then normal-sensitized RBC agglomerates separated at 37 degrees C in the irreversible agglutination-separation (AS) reactions. With separation the AS products. HN spikes (150-200 kDa) and altered NDV, which contain fewer HN spikes than intact allantoic NDV, were removed from the sensitized RBC and the NDV membrane. Extraction of HN spikes from the membrane required more sialic acid than the removal of AS products from RBC. Thus 2 reactions were delineated for the orderly removal. Amounts of each released AS product suggest the source of the HN spikes. AS reactions and ether treatment of NDV increased the HA titres up to 19.2 fold. HA-sialic acid configurations were estimated by the amounts of normal RBC agglutinated by sensitized RBC and also by agglutination with fetuin. Elution of B1 vaccine, HN spikes from ether-treated NDV as well as AS products separated on sephadex resulted in incompletely sensitized RBC; fewer configurations were titrated with normal RBC; all failed to respond to anti-NA antibody. In contrast, sensitized RBC or suspensions of 575 NDV, but not the B1 vaccine strain, responded to both anti-NA and anti-HA antibody. Sensitization, slow elution, and responding to anti-NA antibody, which was accompanied by fluorescent foci on sensitized RBC, required intact NDV and the infrequent HA-sialic acid configuration. The NA was inactive for the HA-sialic acid configuration but cleaved fetuin, indicating substrate specificity. An inactive NA would allow time for fusion and NDV penetration rather than elution by an active NA early in NDV infection.

摘要

575株新城疫病毒（NDV）洗脱后，红细胞（RBC）会被致敏。血凝素（HA）-唾液酸构型中的神经氨酸酶（NA）无活性。致敏红细胞上的HA会凝集正常红细胞。正常红细胞上的唾液酸引发了NA的重新激活——一种新描述的功能。然后，正常致敏红细胞聚集体在37℃下在不可逆凝集分离（AS）反应中分离。随着AS产物的分离，HN刺突（150 - 200 kDa）和改变的NDV（其含有的HN刺突比完整尿囊NDV少）从致敏红细胞和NDV膜中去除。从膜中提取HN刺突比从红细胞中去除AS产物需要更多的唾液酸。因此，确定了2种用于有序去除的反应。每种释放的AS产物的量表明了HN刺突的来源。NDV的AS反应和乙醚处理使HA效价提高了19.2倍。通过致敏红细胞凝集的正常红细胞数量以及与胎球蛋白的凝集来估计HA - 唾液酸构型。B1疫苗的洗脱、经乙醚处理的NDV中的HN刺突以及在葡聚糖凝胶上分离的AS产物导致红细胞致敏不完全；用正常红细胞滴定的构型较少；所有这些都未对抗NA抗体产生反应。相比之下，致敏红细胞或575 NDV悬液，但不是B1疫苗株，对抗NA和抗HA抗体均有反应。致敏、缓慢洗脱以及对抗NA抗体产生反应（致敏红细胞上伴有荧光灶）需要完整的NDV和罕见的HA - 唾液酸构型。NA对HA - 唾液酸构型无活性，但能裂解胎球蛋白，表明底物特异性。无活性的NA将为融合和NDV穿透留出时间，而不是在NDV感染早期被活性NA洗脱。