Ogiso Subaru, Shirai Junsuke, Tuchiya Yoshinori, Honda Eiichi
Tokyo Research and Development Center, Daiichi Pharmaceutical Co. Ltd., Edogawa, Tokyo, Japan.
Uirusu. 2005 Dec;55(2):317-26. doi: 10.2222/jsv.55.317.
Antiviral assay is used routinely for measuring the biological activity of interferon (IFN). However, the challenge viruses used in these assays are considered dangerous to the animal industry and pose a risk of human infection. For example, the vesicular stomatitis virus (VSV) is an important exotic disease agent in domestic animals, and the sindbis virus provokes rash, arthralgia, and fever in humans. Therefore, biosafety needs to be considered when antiviral assays are performed. We chose Getah virus as a candidate challenge virus because it is less hazardous to animals and humans. Crystal violet staining 50% CPE inhibition antiviral assay of human IFN using Getah virus was studied. Antiviral assay using Getah virus and FL cells gave a higher titer of human IFN than did assay using VSV. The titer of human IFN alpha was almost the same as that given by standardized control samples. The titer of human IFN by antiviral assay using Getah virus on the sheet method (IFN reacted the sheeted FL cells) was higher than those of the simultaneous reaction method (IFN reacted the suspending FL cells before sheeted). We therefore consider the sheet method useful for detection of small amounts of IFN. Antiviral assay using Getah virus on MDBK cells gave a lower titer of human IFN alpha than did assay using VSV. However, the adjusting the number of MDBK cells and the titer of Getah virus to get the best condition for CPE appearance, gave similar results in the assays using Getah virus and VSV. We consider that Getah virus is a potentially useful challenge virus for antiviral assay of human IFN.
抗病毒测定法通常用于测量干扰素(IFN)的生物活性。然而,这些测定中使用的攻击病毒被认为对畜牧业有危害,且存在人类感染风险。例如,水疱性口炎病毒(VSV)是家畜中的一种重要外来致病因子,而辛德比斯病毒会引发人类皮疹、关节痛和发热。因此,进行抗病毒测定时需要考虑生物安全性。我们选择盖塔病毒作为候选攻击病毒,因为它对动物和人类的危害较小。研究了使用盖塔病毒对人干扰素进行结晶紫染色50% CPE抑制抗病毒测定。与使用VSV的测定相比,使用盖塔病毒和FL细胞进行的抗病毒测定得到的人干扰素滴度更高。人干扰素α的滴度与标准化对照样品给出的滴度几乎相同。使用盖塔病毒通过片层法(干扰素与铺展的FL细胞反应)进行抗病毒测定得到的人干扰素滴度高于同时反应法(干扰素在铺展前与悬浮的FL细胞反应)。因此,我们认为片层法对于检测少量干扰素很有用。在MDBK细胞上使用盖塔病毒进行抗病毒测定得到的人干扰素α滴度低于使用VSV的测定。然而,调整MDBK细胞数量和盖塔病毒滴度以获得CPE出现的最佳条件,在使用盖塔病毒和VSV的测定中得到了相似的结果。我们认为盖塔病毒对于人干扰素的抗病毒测定是一种潜在有用的攻击病毒。