Murali-Krishna K, Ravi V, Manjunath R
Department of Biochemistry, Indian Institute of Science, India.
Arch Virol. 1995;140(1):127-43. doi: 10.1007/BF01309728.
Ten different mouse cell lines were examined for Japanese encephalitis virus (JEV) infection in vitro and then tested for their ability to generate virus specific cytotoxic T lymphocytes (CTL). Among all cell lines examined, Neuro 2a (a neuroblastoma) was readily infected with JEV as examined by immunofluorescence and viral replication. Among other cells, P388D1, RAW 264.7 (Macrophage origin), Sp2/0 (B-cell Hybridoma), YAC-1 (T-cell lymphoma), and L929 (Fibroblast) were semipermissive to JEV infection. The cytopathic effects caused by progressive JEV infection varied from cell line to cell line. In the case of YAC-1 cells long-term viral antigen expression was observed without significant alterations in cell viability. Intermediate degrees of cytopathicity are seen in RAW 264.7 and L929 cells while infection of PS, Neuro 2a, P388D1 and Sp2/0 caused major viability losses. All infected cell lines were able to prime adult BALB/c (H-2d) mice for the generation of secondary JEV specific CTL. In contrast to YAC-1, the permissive neuroblastoma cell line Neuro 2a (H-2KkDd) was found to be least efficient in its ability to stimulate anti-viral CTL generation. Cold target competition studies demonstrated that both Neuro 2a and YAC-1 (H-2KkDd) cells expressed similar viral determinants that are recognised by CTL, suggesting that the reason for the lower ability of Neuro 2a to stimulate anti-viral CTL was not due to lack of viral CTL determinants. These findings demonstrate that a variety of mouse cell lines can be infected with Japanese encephalitis virus, and that these infected cells could be utilised to generate virus specific CTL in BALB/c mice.
对十种不同的小鼠细胞系进行了体外日本脑炎病毒(JEV)感染检测,然后检测它们产生病毒特异性细胞毒性T淋巴细胞(CTL)的能力。在所有检测的细胞系中,通过免疫荧光和病毒复制检测发现,Neuro 2a(一种神经母细胞瘤)很容易被JEV感染。在其他细胞中,P388D1、RAW 264.7(巨噬细胞来源)、Sp2/0(B细胞杂交瘤)、YAC-1(T细胞淋巴瘤)和L929(成纤维细胞)对JEV感染呈半容许性。进行性JEV感染引起的细胞病变效应因细胞系而异。在YAC-1细胞中,观察到长期病毒抗原表达,而细胞活力无明显改变。RAW 264.7和L929细胞出现中度细胞病变,而PS、Neuro 2a、P388D1和Sp2/0细胞感染则导致细胞活力大幅丧失。所有感染的细胞系都能够使成年BALB/c(H-2d)小鼠致敏,以产生继发性JEV特异性CTL。与YAC-1相反,容许性神经母细胞瘤细胞系Neuro 2a(H-2KkDd)在刺激抗病毒CTL产生的能力方面效率最低。冷靶竞争研究表明,Neuro 2a和YAC-1(H-2KkDd)细胞都表达类似的被CTL识别的病毒决定簇,这表明Neuro 2a刺激抗病毒CTL能力较低的原因不是缺乏病毒CTL决定簇。这些发现表明,多种小鼠细胞系可被日本脑炎病毒感染,并且这些感染细胞可用于在BALB/c小鼠中产生病毒特异性CTL。