Rivetz B, Lipkind M
Arch Virol. 1985;85(3-4):231-55. doi: 10.1007/BF01314234.
Nine NDV strains belonging to lentogenic, mesogenic and velogenic groups were studied. Virus adsorption to chicken red blood cell (RBC) surface was performed at 4 degrees C, and after a temperature shift from 4 degrees to 37 degrees C elution of pre-adsorbed virus and accumulation of free N-acetyl-neuraminic acid (NANA) split from RBC receptors as a result of neuraminidase (Nase) activity was detected. In the case of high multiplicity of adsorption the elution was very fast (complete elution within 5 minutes) for all the strains irrespective of their virulence. Although physical saturation of RBC surface with the adsorbed virus was not achieved, a certain minimal (strain-specific) amount of the pre-adsorbed virus which splits a maximally possible (for a given strain) quantity of the NANA was found (a state of "enzymatic saturation"). Below a certain low multiplicity of adsorption elution was delayed for about 20-30 minutes while the accumulation of the split NANA began immediately after the temperature shift. This phenomenon was interpreted as a result of "crawling" of the adsorbed virions upon the RBC surface followed by "browsing" of RBC receptors and liberation of NANA. Thus, the Nase activity of the attached virus ("in situ Nase activity") is a factor providing both elution and "crawling" of the virus (depending on the multiplicity of adsorption). The in situ Nase activity of all the strains used was determined quantitatively by (1) parameters of enzymatic kinetics (Vmax, Km and Km/Vmax) and (2) parameters of enzymatic efficiency related to a certain quantity of the adsorbed virus, namely, per amount of: a) "crawling" virus, b) that providing "enzymatic saturation", and c) that equal to Km. Computation of these parameters revealed inverse correlation between the in situ Nase activity and the strain virulence. Thus, these indications can be in vitro markers of the in vivo virulence.
对9株属于缓发型、中等毒力型和速发型的新城疫病毒(NDV)毒株进行了研究。在4℃下进行病毒对鸡红细胞(RBC)表面的吸附,在温度从4℃转变为37℃后,检测到预吸附病毒的洗脱以及由于神经氨酸酶(Nase)活性从RBC受体上裂解下来的游离N - 乙酰神经氨酸(NANA)的积累。在高吸附复数的情况下,所有毒株的洗脱都非常快(5分钟内完全洗脱),与它们的毒力无关。尽管未实现RBC表面被吸附病毒的物理饱和,但发现了一定量的预吸附病毒(毒株特异性),其裂解了最大可能量(对于给定毒株)的NANA(“酶促饱和”状态)。在低于一定的低吸附复数时,洗脱延迟约20 - 30分钟,而裂解的NANA在温度转变后立即开始积累。这种现象被解释为吸附的病毒粒子在RBC表面“爬行”,随后“浏览”RBC受体并释放NANA的结果。因此,附着病毒的Nase活性(“原位Nase活性”)是一个决定病毒洗脱和“爬行”的因素(取决于吸附复数)。所使用的所有毒株的原位Nase活性通过以下方式进行定量测定:(1)酶动力学参数(Vmax、Km和Km/Vmax)和(2)与一定量吸附病毒相关的酶促效率参数,即每量的:a)“爬行”病毒,b)提供“酶促饱和”的病毒,以及c)等于Km的病毒。这些参数的计算揭示了原位Nase活性与毒株毒力之间的负相关。因此,这些指标可作为体内毒力的体外标志物。