PATHOGENICITY AND IMMUNOGENICITY FOR SOME RABBIT HEMORRHAGIC DISEASE VIRUS(RHDV) STRAINS PROPAGATED IN ISOLATED HEPATOCYTES OF RABBIT

Yong-Mi Pyon, Hyok Ryo, Sun-Nam Pak, Il-Ryong Ko
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Abstract

The hepatocytes were isolated from the liver of different week-age of healthy rabbits by a single two-stage pass perfusion method and suspended in Hanks’ balanced salt solution (Ca[Formula: see text]-, Mg[Formula: see text]-free). The field strains of RHDV were inoculated into the suspension and incubated for 2 days at 37∘C. After incubation, the HA titer of each strain of RHDV in supernatant of virus growth was measured using human erythrocyte O type. There were differences in the HA titers of RHDV strains according to the hepatocyte of different week-age of rabbits, the number of hepatocyte, the strain of virus (RHDV-1, RHDV-2, RHDV-3) and the duration of incubation. The HA titer was highest when the virus was cultured in the hepatocytes isolated from more than 12 week-aged rabbits. When non-immune rabbits were inoculated with the strains of virus (RHDV-1, RHDV-2, RHDV-3). respectively, all rabbits (3 heads of rabbit per group) infected with RHDV-1 and RHDV-3 died of the disease within 48[Formula: see text]h, while only one rabbit infected with RHDV-2 died within 72[Formula: see text]h. When immunized rabbits by inactivated virus strains (RHDV-1, RHDV-2 and RHDV-3), 2 strains, RHDV-1 and RHDV-3, showed the same level of HI titers with inactivated homogenate, but RHDV-2 did lower the level of titer than the homogenate.
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兔出血性疾病病毒(rhdv)在兔肝细胞中的致病性和免疫原性
采用单级两段式灌注法从不同周龄的健康家兔肝脏中分离肝细胞,悬浮于Hanks平衡盐溶液中(Ca[公式:见文]-,Mg[公式:见文]-)。将RHDV野外菌株接种到悬浮液中,在37°C下孵育2天。孵育后,采用人红细胞O型法测定各株RHDV在病毒生长上清液中的HA滴度。不同周龄家兔的肝细胞、肝细胞数、病毒株(RHDV-1、RHDV-2、RHDV-3)和孵育时间不同,不同病毒株的HA滴度也不同。当病毒在12周龄以上家兔肝细胞中培养时,HA滴度最高。将无免疫力的家兔接种病毒株(RHDV-1、RHDV-2、RHDV-3)。分别感染RHDV-1和RHDV-3的家兔(每组3头)在48小时内全部死亡[公式:见文],而感染RHDV-2的家兔在72小时内只有1只死亡[公式:见文]。用灭活病毒株(RHDV-1、RHDV-2和RHDV-3)免疫家兔时,2株(RHDV-1和RHDV-3)的HI滴度与灭活匀浆相同,但RHDV-2的滴度低于匀浆。
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