Extraintestinal Spread and Replication of a Homologous EC Rotavirus Strain and a Heterologous Rhesus Rotavirus in BALB/c Mice

Abstract

Although rotavirus infection has generally been felt to be restricted to the gastrointestinal tract, over the last two decades there have been sporadic reports of children with acute or fatal cases of rotavirus gastroenteritis testing positive for rotavirus antigen and/or nucleic acid in various extraintestinal locations such as serum, liver, kidney, bladder, testes, nasal secretions, cerebrospinal fluid, and the central nervous system. Recently, studies in animals and people have demonstrated that rotavirus antigenemia is a common event during natural infection. In this study, we extend these observations and compare the intestinal and extraintestinal spread of wild-type homologous murine rotavirus EC and a heterologous strain, rhesus rotavirus (RRV), in newborn mice. A strand-specific quantitative reverse transcription-PCR (ssQRT-PCR) assay was used to quantify the ability of different rotavirus strains to spread and replicate extraintestinally. Both strain EC and RRV were detected extraintestinally in the mesenteric lymph nodes (MLN), livers, lungs, blood, and kidneys. Extraintestinal replication, as measured by ssQRT-PCR, was most prominent in the MLN and occurred to a lesser degree in the livers, kidneys, and lungs. In the MLN, strain EC and RRV had similar (P < 0.05) RNA copy numbers, although EC was present at a 10,000-fold excess over RRV in the small intestine. Rotavirus nonstructural protein 4 (NSP4) and/or assembled triple-layered particles, indicated by immunostaining with the VP7 conformation-dependent monoclonal antibody 159, were detected in the MLN, lungs, and livers of ECand RRV-inoculated mice, confirming the ssQRT-PCR findings. Infectious RRV was detected in the MLN in quantities exceeding the amount present in the small intestines or blood. The cells in the MLN that supported rotavirus replication included dendritic cells and potentially B cells and macrophages. These data indicate that extraintestinal spread and replication occurs commonly during homologous and some heterologous rotaviral infections; that the substantial host range restrictions for rhesus rotavirus, a heterologous strain present in the intestine, are not necessarily apparent at systemic sites; that the level and location of extraintestinal replication varies between strains; that replication can occur in several leukocytes subsets; and that extraintestinal replication is likely a part of the normal pathogenic sequence of homologous rotavirus infection.Rotavirus-associated gastroenteritis affects an estimated 111 million children worldwide and is responsible for at least 440,000 deaths annually (25). Although in some reports rotavirus antigen and/or RNA was detected in the central nervous systems, lungs, kidneys, spleens, heart, testes, bladders, and pancreases of selected severely ill children (19,20), homologous rotavirus replication has generally been considered to be restricted to the terminally differentiated epithelial cells of the small intestinal villi. The sporadically reported findings of extraintestinal virus have ...

Persistence of the rotavirus A genome in mesenteric lymph nodes of cattle raised on farms

“…Although we were not able to exclude the possibility that the negative result for PCR in intestinal contents could be due to low viral titre in the digestive tract, the data are consistent with the idea that viral genomes become present in MLNs at a late stage of infection. Furthermore, a previous study using a mouse model showed that replication of the simian RRV strain persists longer in the MLNs than in the digestive tract (Fenaux et al, 2006). Consistent with this, our data indicated the possibility that viral genomes can persist in MLNs after viral clearance in the gut under natural conditions, implying that RVAs had persisted in MLNs of cattle for prolonged periods.…”

“…The presence of RVA in MLNs has so far been reported in experimental animal models (Brown & Offit, 1998;Crawford et al, 2006;Dharakul et al, 1988;Fenaux et al, 2006;Kim et al, 2011;Mossel & Ramig, 2003; Park et al, Genetic analyses showed that all strains for which the genome was detected in MLNs from autopsied calves and healthy adult cattle were classified as G and/or P (possible) genotypes generally prevalent in bovines (Tables 2 and 3). Also, in most of the cattle that had RVA genomes in both MLNs and intestinal contents, the strains in intestinal contents were genetically identical to those in MLNs.…”

“…Notably, a previous study using a mouse model showed that RVA strain RRV persists longer in the MLNs than in the small intestine (Fenaux et al, 2006). Also, previous studies on the dynamics of RVA in extra-intestinal organs in calves under experimental conditions showed that MLNs had the highest viral RNA copy number amongst extra-intestinal organs (Kim et al, 2011;Park et al, 2013Park et al, , 2014.…”

“…However, there has been an increasing number of reports of infectious RVA and the viral antigen being detected in sera and various organs, such as mesenteric lymph nodes (MLNs), lungs and livers in humans and animals (Blutt et al, 2003;Crawford et al, 2006;Dharakul et al, 1988;Fenaux et al, 2006;Lynch et al, 2003;Mossel & Ramig, 2003). In experimental animal models, RVA has been detected not only in the gastrointestinal tract but also in extra-intestinal organs, indicating that the virus is able to spread systemically from the gastrointestinal tract (Crawford et al, 2006;Fenaux et al, 2006;Kim et al, 2011;Mossel & Ramig, 2003;Park et al, 2013Park et al, , 2014.…”

Rotavirus acceleration of murine type 1 diabetes is associated with a T helper 1-dependent specific serum antibody response and virus effects in regional lymph nodes

“…Similarly, MLN in infected infants of diabetes-resistant mice harbour infectious RRV. The cells in the MLN involved are unidentified, although mouse rotavirus replicates in dendritic cells (DC), and possibly B cells and macrophages [21,22]. The spread of rotavirus to the MLN and PLN in adult mice has not been reported.…”