Stringent Requirement for the C Protein of Wild-Type Measles Virus for Growth both In Vitro and in Macaques
Stringent Requirement for the C Protein of Wild-Type Measles Virus for Growth both In Vitro and in Macaques
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DOI：
10.1128/jvi.79.12.7838-7844.2005
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发表时间：
2005-06
影响因子：
5.4
通讯作者：
K. Takeuchi;M. Takeda;N. Miyajima;Y. Ami;N. Nagata;Y. Suzaki;Jamila Shahnewaz;Shin-ichi Kadota;K. Nagata
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文献类型：
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作者：
K. Takeuchi;M. Takeda;N. Miyajima;Y. Ami;N. Nagata;Y. Suzaki;Jamila Shahnewaz;Shin-ichi Kadota;K. Nagata
麻疹病毒（MV）P基因编码P蛋白和三种辅助蛋白（C、V和R）。然而，这些辅助蛋白在MV感染的自然过程中的作用仍不清楚。在这项研究中，我们通过使用反向遗传学系统（M.武田，K. Takeuchi，N. Miyajima，F. Kobune，Y. Ami，N. Nagata，Y. Suzaki，Y.永井和M.田代，J. 74：6643-6647）。当表达MV受体SLAM（293/hSLAM）的293细胞被wtMV（C-）或亲本野生型MV（wtMV）感染时，wtMV（C-）的生长受到限制，特别是在后期。增强的绿色荧光蛋白表达的wtMV（C−）一致诱导晚期细胞变圆和细胞死亡的融合抑制肽的存在下，表明C蛋白可以防止细胞死亡，并需要长期MV感染。抗α/β干扰素的中和抗体不能恢复293/hSLAM细胞中wtMV（C−）的生长限制。当食蟹猴感染wtMV（C−）或wtMV时，胸腺中MV感染细胞的数量wtMV（C−）比wtMV少1，000倍以上。免疫组织化学分析显示，在感染wtMV的食蟹猴的脾脏、淋巴结、扁桃体和喉中，MV抗原表达强烈，但在感染wtMV的食蟹猴的相同组织中，表达显著降低（C−）。这些数据表明MV C蛋白对于体外和食蟹猴中的有效MV复制是必需的。
ABSTRACT The P gene of measles virus (MV) encodes the P protein and three accessory proteins (C, V, and R). However, the role of these accessory proteins in the natural course of MV infection remains unclear. For this study, we generated a recombinant wild-type MV lacking the C protein, called wtMV(C−), by using a reverse genetics system (M. Takeda, K. Takeuchi, N. Miyajima, F. Kobune, Y. Ami, N. Nagata, Y. Suzaki, Y. Nagai, and M. Tashiro, J. Virol. 74:6643-6647). When 293 cells expressing the MV receptor SLAM (293/hSLAM) were infected with wtMV(C−) or parental wild-type MV (wtMV), the growth of wtMV(C−) was restricted, particularly during late stages. Enhanced green fluorescent protein-expressing wtMV(C−) consistently induced late-stage cell rounding and cell death in the presence of a fusion-inhibiting peptide, suggesting that the C protein can prevent cell death and is required for long-term MV infection. Neutralizing antibodies against alpha/beta interferon did not restore the growth restriction of wtMV(C−) in 293/hSLAM cells. When cynomolgus monkeys were infected with wtMV(C−) or wtMV, the number of MV-infected cells in the thymus was >1,000-fold smaller for wtMV(C−) than for wtMV. Immunohistochemical analyses showed strong expression of an MV antigen in the spleen, lymph nodes, tonsils, and larynx of a cynomolgus monkey infected with wtMV but dramatically reduced expression in the same tissues in a cynomolgus monkey infected with wtMV(C−). These data indicate that the MV C protein is necessary for efficient MV replication both in vitro and in cynomolgus monkeys.