ABSTRACT

Inclusion body nephropathy (IBN) and kidney fibrosis in aged immunodeficient mice and, to lesser extent, in immunocompetent mice have been recently linked to infection of mouse kidney parvovirus (MKPV), also known as murine chapparvovirus (MuCPV). Knowledge about its prevalence and the complete genome sequence of more MKPV strains is essential for understanding phylogenetic relationships and pathogenicity among MKPV strains. In the present study using PCR and genome walking, we determined the complete 4440-nucleotide genome of a new MKPV strain, namely MIT-WI1, which was identified in IBN-affected Il2rg^−/–Rag2^−/– c-Kit ^{W–sh/W–sh} mice housed in the vivarium at Whitehead Institute for Biomedical Research (WI). The overall nucleotide (>94%) and deduced amino acid sequences (>98%) of p10, p15, NS1 (replicase), NS2 and VP1 (capsid protein) within the MIT-WI1 genome, are closely related to MKPV/MuCPV strains described in laboratory and wild Mus musculus mice. In addition, PCR and qPCR assays using newly designed primers conserved among the known MKPV/MuCPV genomes were developed and utilized to assess MKPV status in selected laboratory mice. MKPV was also detected in immunodeficient (NSG) and immunocompetent (Crl:CD1(ICR), UTX^flox) mouse strains/stocks. The abundance of the MKPV genome copies was significantly correlated with the severity of IBN. Our data indicate that MKPV is present in selected mouse strains/stocks, and provides new insights into the genome evolution of MKPV.

摘要

近来，在免疫缺陷的老龄小鼠和免疫功能较弱的小鼠中，包涵体肾病（IBN）和肾纤维化已与小鼠肾脏细小病毒（MKPV）（也称为鼠类细小病毒（MuCPV））的感染有关。有关其流行性以及更多MKPV菌株的完整基因组序列的知识对于理解MKPV菌株之间的系统发生关系和致病性至关重要。在使用PCR和基因组步移的本研究中，我们确定了新MKPV菌株MIT-WI1的完整4440个核苷酸的基因组，该菌株在IBN感染的Il2rg ^-/- Rag2 ^-/- c-Kit ^W-sh中鉴定出^{/ W–sh} 怀特黑德生物医学研究所（WI）的动物饲养箱中的小鼠。MIT-WI1基因组中p10，p15，NS1（复制酶），NS2和VP1（衣壳蛋白）的总核苷酸（> 94％）和推导的氨基酸序列（> 98％）与MKPV / MuCPV菌株密切相关描述于实验室和野生小家鼠小鼠中。此外，开发了使用新设计的已知MKPV / MuCPV基因组中保守的引物进行的PCR和qPCR分析，并将其用于评估所选实验室小鼠中MKPV的状态。在免疫缺陷（NSG）和具有免疫能力的（Crl：CD1（ICR），UTX ^flox中也检测到^MKPV）小鼠品系/种群。MKPV基因组拷贝的丰度与IBN的严重程度显着相关。我们的数据表明，MKPV存在于选定的小鼠品系/种群中，并为MKPV的基因组进化提供了新的见解。