Disruption of the M2 gene of murine gammaherpesvirus 68 alters splenic latency following intranasal, but not intraperitoneal, inoculation
Disruption of the M2 gene of murine gammaherpesvirus 68 alters splenic latency following intranasal, but not intraperitoneal, inoculation
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DOI：
10.1128/jvi.76.4.1790-1801.2002
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发表时间：
2002-02-01
影响因子：
5.4
通讯作者：
Speck, SH
中科院分区：
文献类型：
--
作者：
Jacoby, MA;Virgin, HW;Speck, SH
用鼠γ疱疹病毒68（γ HV 68;也称为MHV 68）感染小鼠提供了一种易于处理的小动物模型，用该模型来解决γ疱疹病毒感染在体内建立和维持的要求。γ HV 68的M2基因是潜伏相关基因，其编码与任何已知病毒或细胞蛋白缺乏可辨别同源性的蛋白。在潜伏感染的脾细胞（S. M. Husain，E. J. Usherwood，H.戴森角Coleclough，M. A. Coppola，D. L.伍德兰，M。A. Blackman，J.P. Stewart，和J.T.样品，Proc. Natl. Acad. Sci. USA 96：7508-7513，1999; H. W.维珍四号河M.普雷斯蒂，X。Y. Li，C. Liu和S. H. Speck，J. Virol. 73：2321-2332，1999）和腹膜渗出细胞（H. W.维珍四号河M.普雷斯蒂，X。Y. Li，C. Liu和S. H. Speck，J. Virol. 73：2321-2332，1999），以及潜伏性γ HV 68感染的B淋巴瘤细胞系（S. M. Husain，E. J. Usherwood，H.戴森角Coleclough，M. A. Coppola，D. L.伍德兰，M。A. Blackman，J.P. Stewart，和J.T.样品，Proc. Natl. Acad. Sci. USA 96：7508-7513，1999）。在这里，我们描述了一代的γ HV 68突变体与破坏的M2基因。M2基因突变不影响病毒在组织培养中复制的能力，也不影响B6.Rag1缺陷小鼠中的γ HV 68毒力。然而，我们发现，M2的差异所需的急性复制在体内。虽然M2突变不影响鼻内接种后小鼠肺中病毒复制的急性期，但与腹膜内接种后野生型和标记拯救病毒相比，脾脏中的急性期病毒复制减少。鼻内接种后，M2突变病毒表现出显着减少建立潜伏期在脾脏感染后第16天，作为测量的病毒基因组阳性细胞的频率。此外，与野生型和标记拯救病毒相比，M2突变病毒基因组阳性细胞从潜伏期重新激活效率低下。到鼻内接种后第42天，M2突变型和野生型病毒基因组阳性细胞的频率几乎相等，并且在两个群体中均检测到很少的再活化。与鼻内接种后获得的结果形成鲜明对比的是，腹膜内接种后，在M2突变病毒的建立或从潜伏期的再激活中未观察到显著缺陷。这些结果表明，建立潜伏期的要求受到感染途径的影响。
Infection of mice with murine gammaherpesvirus 68 (gammaHV68; also referred to as MHV68) provides a tractable small-animal model with which to address the requirements for the establishment and maintenance of gammaherpesvirus infection in vivo. The M2 gene of gammaHV68 is a latency-associated gene that encodes a protein lacking discernible homology to any known viral or cellular proteins. M2 gene transcripts have been detected in latently infected splenocytes (S. M. Husain, E. J. Usherwood, H. Dyson, C. Coleclough, M. A. Coppola, D. L. Woodland, M. A. Blackman, J. P. Stewart, and J. T. Sample, Proc. Natl. Acad. Sci. USA 96:7508-7513, 1999; H. W. Virgin IV, R. M. Presti, X. Y. Li, C. Liu, and S. H. Speck, J. Virol. 73:2321-2332, 1999) and peritoneal exudate cells (H. W. Virgin IV, R. M. Presti, X. Y. Li, C. Liu, and S. H. Speck, J. Virol. 73:2321-2332, 1999), as well as in a latently gammaHV68-infected B-lymphoma cell line (S. M. Husain, E. J. Usherwood, H. Dyson, C. Coleclough, M. A. Coppola, D. L. Woodland, M. A. Blackman, J. P. Stewart, and J. T. Sample, Proc. Natl. Acad. Sci. USA 96:7508-7513, 1999). Here we describe the generation of gammaHV68 mutants with disruptions in the M2 gene. Mutation of the M2 gene did not affect the ability of the virus to replicate in tissue culture, nor did it affect gammaHV68 virulence in B6.Rag1 deficient mice. However, we found that M2 was differentially required for acute replication in vivo. While mutation of M2 did not affect acute phase of virus replication in the lungs of mice following intranasal inoculation, acute-phase virus replication in the spleen was decreased compared to that of the wild-type and marker rescue viruses following intraperitoneal inoculation. Upon intranasal inoculation, M2 mutant viruses exhibited a significant decrease in the establishment of latency in the spleen on day 16 postinfection, as measured by the frequency of viral genome-positive cells. In addition, M2 mutant viral genome-positive cells reactivated from latency inefficiently compared to wild-type and marker rescue viruses. By day 42 after intranasal inoculation, the frequencies of M2 mutant and wild-type viral genome-positive cells were nearly equivalent and little reactivation was detected from either population. In sharp contrast to the results obtained following intranasal inoculation, after intraperitoneal inoculation, no significant defect was observed in the establishment or reactivation from latency with the M2 mutant viruses. These results indicate that the requirements for the establishment of latency are affected by the route of infection.