ABSTRACT
Influenza viruses are seasonally recurring human pathogens. Vaccines and antiviral drugs are available for influenza. However, the viruses, which often change themselves via antigenic drift and shift, demand constant efforts to update vaccine antigens every year and develop new agents with broad-spectrum antiviral efficacy. An animal model is critical for such efforts. While most human influenza viruses are unable to kill BALB/c mice, some strains have been shown to kill DBA/2 mice without prior adaptation. Therefore, in this study, we explored the feasibility of employing DBA/2 mice as a model in the development of anti-influenza drugs. Unlike the BALB/c strain, DBA/2 mice were highly susceptible and could be killed with a relatively low titer (50% DBA/2 lethal dose = 10(2.83) plaque-forming units) of the A/Korea/01/2009 virus (2009 pandemic H1N1 virus). When treated with a neuraminidase inhibitor, oseltamivir phosphate, infected DBA/2 mice survived until 14 days post-infection. The reduced morbidity of the infected DBA/2 mice was also consistent with the oseltamivir treatment. Taking these data into consideration, we propose that the DBA/2 mouse is an excellent animal model to evaluate antiviral efficacy against influenza infection and can be further utilized for combination therapies or bioactivity models of existing and newly developed anti-influenza drugs.
Subject(s)
Antiviral Agents/administration & dosage , Disease Models, Animal , Influenza A Virus, H1N1 Subtype/drug effects , Influenza, Human/drug therapy , Influenza, Human/virology , Mice, Inbred DBA , Animals , Drug Evaluation, Preclinical , Female , Humans , Influenza A Virus, H1N1 Subtype/pathogenicity , Mice , Mice, Inbred BALB C , Mice, Inbred DBA/virology , Republic of Korea , Virulence/drug effectsABSTRACT
Despite the prevalence of H5N1 influenza viruses in global avian populations, comparatively few cases have been diagnosed in humans. Although viral factors almost certainly play a role in limiting human infection and disease, host genetics most likely contribute substantially. To model host factors in the context of influenza virus infection, we determined the lethal dose of a highly pathogenic H5N1 virus (A/Hong Kong/213/03) in C57BL/6J and DBA/2J mice and identified genetic elements associated with survival after infection. The lethal dose in these hosts varied by 4 logs and was associated with differences in replication kinetics and increased production of proinflammatory cytokines CCL2 and tumor necrosis factor alpha in susceptible DBA/2J mice. Gene mapping with recombinant inbred BXD strains revealed five loci or Qivr (quantitative trait loci for influenza virus resistance) located on chromosomes 2, 7, 11, 15, and 17 associated with resistance to H5N1 virus. In conjunction with gene expression profiling, we identified a number of candidate susceptibility genes. One of the validated genes, the hemolytic complement gene, affected virus titer 7 days after infection. We conclude that H5N1 influenza virus-induced pathology is affected by a complex and multigenic host component.
Subject(s)
Genetic Variation , Host-Pathogen Interactions , Influenza A Virus, H5N1 Subtype/pathogenicity , Mice, Inbred C57BL/virology , Mice, Inbred DBA/virology , Orthomyxoviridae Infections/mortality , Animals , Chemokine CCL2/genetics , Chemokine CCL2/metabolism , Chromosome Mapping , Disease Models, Animal , Disease Susceptibility , Female , Gene Expression Profiling , Humans , Influenza A Virus, H5N1 Subtype/genetics , Influenza, Human/mortality , Influenza, Human/virology , Mice/virology , Mice, Inbred C57BL/genetics , Mice, Inbred DBA/genetics , Mice, Mutant Strains , Orthomyxoviridae Infections/genetics , Orthomyxoviridae Infections/virology , Quantitative Trait Loci , Recombination, Genetic , Species Specificity , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolismABSTRACT
BACKGROUND AND PURPOSE: Detection of mouse parvovirus 1 (MPV) depends on use of serologic and polymerase chain reaction (PCR) assays. These assays were evaluated for their ability to detect virus-specific antibodies or viral DNA in multiple strains and ages of mice inoculated with MPV. METHODS: Twelve-week-old ICR, BALB/c, C3H, C57BL/6, and DBA/2 mice and four- and eight-week-old ICR mice were inoculated with MPV. Serum was harvested four weeks after inoculation and analyzed by use of recombinant non structural protein 1 (rNS1) enzyme-linked immunosorbent assay (ELISA), minute virus of mice (MVM) ELISA, and MPV indirect fluorescent antibody (IFA), MVM IFA, and MPV hemagglutination inhibition (HAI) assays. Select tissues were harvested and analyzed by use of an MPV-specific PCR assay. RESULTS: The number of mice in each group with detectable MPV-specific antibodies or MPV DNA varied with mouse strain, mouse age when inoculated, and viral dose. Seroconversion in mice inoculated at 12 weeks of age was detected almost exclusively by use of the MPV IFA and MPV HAI assays, whereas seroconversion in almost all mice inoculated at 4 and 8 weeks of age was detected by use of all immunoassays except the MVM ELISA. Viral DNA was detected by use of PCR analysis in all strains and ages of mice except DBA/2 mice. CONCLUSIONS: Mouse strain and age have important roles in seroconversion to nonstructural and structural MPV antigens and persistence of viral DNA in mouse tissues. Therefore, diagnostic serologic testing and PCR analysis should be considered within the context of mouse strain and age at the time of MPV exposure, especially when sentinel mice are used for surveillance.
Subject(s)
Mice, Inbred Strains/virology , Parvoviridae Infections/veterinary , Parvovirus/isolation & purification , Rodent Diseases/diagnosis , Animals , Antibodies, Viral/blood , DNA Primers/chemistry , DNA, Viral/chemistry , DNA, Viral/isolation & purification , Electrophoresis, Agar Gel/veterinary , Enzyme-Linked Immunosorbent Assay/veterinary , Fluorescent Antibody Technique, Indirect/veterinary , Hemagglutination Inhibition Tests/veterinary , Male , Mice , Mice, Inbred BALB C/virology , Mice, Inbred C3H/virology , Mice, Inbred C57BL/virology , Mice, Inbred DBA/virology , Mice, Inbred ICR/virology , Parvoviridae Infections/diagnosis , Parvoviridae Infections/prevention & control , Parvovirus/genetics , Parvovirus/immunology , Polymerase Chain Reaction/veterinary , Rodent Diseases/prevention & control , Viral Nonstructural Proteins/bloodABSTRACT
We previously described an endogenous murine retrovirus, rv-DBA/2aged, isolated from an aged DBA/2 mouse. The previous report showed that a recombination which resulted in the replacement of Emv-3 gag sequences with gag sequences homologous to those found in the AKT-8 virus had taken place. This recombination allowed production of a competent virus from the defective Emv-3 locus. However, the extent of replacement of Emv-3 gag was not known. We report here the entire sequence for the gag gene of rv-DBA/2aged as well as the previously unsequenced 3' end of the Emv-3 gag gene. These data demonstrate that while sequences homologous to the entire gag gene fragment found in AKT-8 are represented in rv-DBA/2aged, the remainder of rv-DBA/2aged gag is not derived from Emv-3 but is a unique gag sequence. Furthermore, a complete comparison of env sequences shows that the env of rv-DBA/2aged is derived entirely from Emv-3. Additional data suggest that the recombination which led to production of the rv-DBA/2aged virus may be a common event in aging DBA/2 mice. Finally, comparison of the new sequences of Emv-3 with those of the Akv virus (also designated AKR-623 and Emv-11) and Emv-1 shows that this endogenous virus locus is very closely related to the other Emv loci at the nucleotide sequence level.
