Kimman T G, Bianchi A T, de Bruin T G, Mulder W A, Priem J, Voermans J J
Department of Virology, Central Veterinary Institute, Lelystad, The Netherlands.
Vet Immunol Immunopathol. 1995 Apr;45(3-4):253-63. doi: 10.1016/0165-2427(94)05344-r.
We examined whether the L14 cell line, an immortalized B cell line originating from inbred miniature pigs of the MHC haplotype d/d, could be useful to study T cell responses of pigs to pseudorabies virus (PRV). Compared with porcine kidney cells, the replication of PRV in L14 cells was slower and yielded lower quantities of infectious virus, which agrees with the reported poor replication of PRV in peripheral blood lymphocytes of swine. The virus yield and the number of L14 cells expressing the viral glycoprotein gE were both maximal at 48 h postinfection, when approximately 90% of all viable L14 cells expressed gE. Morphologically detectable effects of PRV replication in L14 cells were not obvious, but the number of viable cells at 72 h postinfection was lower in infected cultures than in uninfected cultures. Major histocompatibility complex (MHC) class I and II antigen expression was significantly higher at different time points postinfection on infected than on uninfected L14 cells. In contrast, expression of IgM appeared very slightly reduced on infected L14 cells, indicating a selective influence of PRV on cellular protein expression. PRV-infected L14 cells were lysed by lymphocytes from PRV-immune minipigs of MHC haplotype d/d, indicating their usefulness in in vitro cytolytic assays.
我们研究了源自MHC单倍型为d/d的近交系小型猪的永生化B细胞系L14,是否可用于研究猪对伪狂犬病病毒(PRV)的T细胞反应。与猪肾细胞相比,PRV在L14细胞中的复制较慢,产生的感染性病毒量较低,这与报道的PRV在猪外周血淋巴细胞中复制不佳一致。感染后48小时,病毒产量和表达病毒糖蛋白gE的L14细胞数量均达到最大值,此时所有存活的L14细胞中约90%表达gE。PRV在L14细胞中复制的形态学可检测效应不明显,但感染后72小时,感染培养物中的活细胞数量低于未感染培养物。感染后不同时间点,感染的L14细胞上主要组织相容性复合体(MHC)I类和II类抗原表达显著高于未感染的L14细胞。相反,感染的L14细胞上IgM的表达似乎略有降低,表明PRV对细胞蛋白表达有选择性影响。PRV感染的L14细胞被来自MHC单倍型为d/d的PRV免疫小型猪的淋巴细胞裂解,表明它们在体外细胞溶解试验中有用。