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通过杆状病毒载体表达产生的犬细小病毒空衣壳:用于病毒特性分析和犬类免疫接种

Canine parvovirus empty capsids produced by expression in a baculovirus vector: use in analysis of viral properties and immunization of dogs.

作者信息

Saliki J T, Mizak B, Flore H P, Gettig R R, Burand J P, Carmichael L E, Wood H A, Parrish C R

机构信息

James A. Baker Institute, New York State College of Veterinary Medicine, Cornell University, Ithaca 14853.

出版信息

J Gen Virol. 1992 Feb;73 ( Pt 2):369-74. doi: 10.1099/0022-1317-73-2-369.

Abstract

The VP-2 genes of canine parvovirus (CPV) and a recombinant consisting of CPV and feline panleukopenia virus (FPV) sequences were cloned into baculovirus expression vectors, fused to the baculovirus polyhedrin promoter. Recombinant baculoviruses were prepared and the properties of the parvovirus proteins expressed in insect cells examined. The proteins produced were the same size as the authentic CPV VP-2 protein, and were produced late after infection; the quantity of proteins recovered from the insect cell cultures was similar to those produced in CPV infections. Parvovirus particles formed had the haemagglutination (HA), sedimentation and buoyant density properties of authentic CPV capsids. Both the CPV capsids and the CPV-FPV recombinant capsids from the baculovirus system expressed the same epitopes as those seen in the viable parvoviruses when tested with a panel of anti-parvovirus monoclonal antibodies. Lysates of recombinant baculovirus-infected cells were inoculated into dogs, giving rise to serum neutralizing and HA-inhibiting antibodies, and the immunized dogs were protected from clinical disease upon challenge with a virulent isolate of the most recent antigenic type of CPV.

摘要

将犬细小病毒(CPV)的VP - 2基因以及由CPV和猫泛白细胞减少症病毒(FPV)序列组成的重组体克隆到杆状病毒表达载体中,与杆状病毒多角体蛋白启动子融合。制备重组杆状病毒,并检测在昆虫细胞中表达的细小病毒蛋白的特性。所产生的蛋白与天然CPV VP - 2蛋白大小相同,且在感染后期产生;从昆虫细胞培养物中回收的蛋白量与CPV感染时产生的量相似。形成的细小病毒颗粒具有天然CPV衣壳的血凝(HA)、沉降和浮力密度特性。当用一组抗细小病毒单克隆抗体检测时,来自杆状病毒系统的CPV衣壳和CPV - FPV重组衣壳与活细小病毒中所见的表达相同的表位。将重组杆状病毒感染细胞的裂解物接种到犬体内,产生血清中和抗体和HA抑制抗体,并且在用最新抗原型CPV的强毒株攻击时,免疫的犬受到保护不患临床疾病。

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