Riviere M, Tartaglia J, Perkus M E, Norton E K, Bongermino C M, Lacoste F, Duret C, Desmettre P, Paoletti E
Rhone Merieux, Lyon, France.
J Virol. 1992 Jun;66(6):3424-34. doi: 10.1128/JVI.66.6.3424-3434.1992.
Glycoproteins gp50, gII, and gIII of pseudorabies virus (PRV) were expressed either individually or in combination by vaccinia virus recombinants. In vitro analysis by immunoprecipitation and immunofluorescence demonstrated the expression of a gII protein of approximately 120 kDa that was proteolytically processed to the gIIb (67- to 74-kDa) and gIIc (58-kDa) mature protein species similar to those observed in PRV-infected cells. Additionally, the proper expression of the 90-kDa gIII and 50-kDa gp50 was observed. All three of these PRV-derived glycoproteins were detectable on the surface of vaccinia virus-PRV recombinant-infected cells. In vivo, mice were protected against a virulent PRV challenge after immunization with the PRV glycoprotein-expressing vaccinia virus recombinants. The coexpression of gII and gIII by a single vaccinia virus recombinant resulted in a significantly reduced vaccination dose required to protect mice against PRV challenge. Inoculation of piglets with the various vaccinia virus-PRV glycoprotein recombinants also resulted in protection against virulent PRV challenge as measured by weight gain. The simultaneous expression of gII and gp50 in swine resulted in a significantly enhanced level of protection as evaluated by weight evolution following challenge with live PRV.
伪狂犬病病毒(PRV)的糖蛋白gp50、gII和gIII通过痘苗病毒重组体单独或组合表达。通过免疫沉淀和免疫荧光进行的体外分析表明,约120 kDa的gII蛋白得以表达,该蛋白经过蛋白水解加工形成gIIb(67至74 kDa)和gIIc(58 kDa)成熟蛋白种类,类似于在PRV感染细胞中观察到的情况。此外,还观察到了90 kDa的gIII和50 kDa的gp50的正确表达。这三种源自PRV的糖蛋白在痘苗病毒-PRV重组体感染细胞的表面均能被检测到。在体内,用表达PRV糖蛋白的痘苗病毒重组体免疫小鼠后,可使其免受强毒PRV攻击。单个痘苗病毒重组体共表达gII和gIII可显著降低保护小鼠免受PRV攻击所需的疫苗接种剂量。用各种痘苗病毒-PRV糖蛋白重组体接种仔猪,通过体重增加衡量,也能使其免受强毒PRV攻击。在猪体内同时表达gII和gp50,通过感染活PRV后的体重变化评估,可显著增强保护水平。