| 1. |
- Ekström, Jens-Ola, et al.
(författare)
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Replication of Ljungan virus in cell culture : the genomic 5'-end, infectious cDNA clones and host cell response to viral infections
- 2007
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Ingår i: Virus Research. - : Elsevier BV. - 0168-1702 .- 1872-7492. ; 130:1-2, s. 129-139
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Tidskriftsartikel (refereegranskat)abstract
- Ljungan virus (LV) is a picornavirus recently isolated from bank voles (Clethrionomys glareolus). The previously uncharacterised 5'-end sequence of the LV genome was determined. Infectious cDNA clones were constructed of the wild type LV prototype strain 87-012 and of the cytolytically replicating cell culture adapted variant 87-012G. Virus generated from cDNA clones showed identical growth characteristics as uncloned virus stocks. Cell culture adapted LV, 87-012G, showed a clear cytopathic effect (CPE) at 3-4 days post-infection (p.i.). Virus titers, determined by plaque titration, increased however only within the first 18h p.i. Replication of LV (+) strand RNA was determined by real-time PCR and corresponded in time with increasing titers. In contrast, the amounts of the replication intermediate, the (-) strand, continued to increase until the cells showed CPE. This indicates separate controlling mechanisms for replication of LV (+) and (-) genome strands. Replication was also monitored by immunofluorescence (IF) staining. IF staining of both prototype 87-012 and the CPE causing 87-012G showed groups of 5-25 infected cells at 48h p.i., suggesting a, for picornaviruses, not previously described direct cell-to-cell transmission.
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| 2. |
- Kallies, Rene, et al.
(författare)
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Development and characterization of murine monoclonal antibodies to first and second Ljungan virus genotypes
- 2012
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Ingår i: Journal of Virological Methods. - : Elsevier BV. - 0166-0934 .- 1879-0984. ; 184:1-2, s. 27-33
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Tidskriftsartikel (refereegranskat)abstract
- Ljungan virus (LV) is a rodent pathogen that causes diabetes and myocarditis in its natural host. In addition, LV has been associated with human disease during pregnancy and of neonates, respectively. A panel of 22 monoclonal antibodies (mAbs) against first and second LV genotypes were produced by immunization of BALB/c mice with whole virus. Thirteen mAbs were class IgG antibodies and nine were class IgM antibodies; all of them contained kappa light chains. All mAbs were reactive with LV by capture enzyme-linked immunosorbent assay and indirect immunofluorescence assay. In addition, five mAbs showed a positive staining in immunohistochemistry. No mAb bound to denatured capsid proteins detected by western immunoblotting. In contrast, the target capsid protein(s) of 20 mAbs were identified by immune precipitation, revealing the conformational nature of epitopes required for mAb binding. None of the mAbs reacted with third and fourth LV genotypes. mAbs characterized should provide useful tools for the development of diagnostic assays and the investigation of LV first and second genotype properties and its pathogenesis.
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| 3. |
- Niklasson, Bo, et al.
(författare)
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Diabetes Prevention Through Antiviral Treatment in Biobreeding Rats
- 2016
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Ingår i: Viral immunology. - : Mary Ann Liebert Inc. - 0882-8245 .- 1557-8976. ; 29:8, s. 452-458
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Tidskriftsartikel (refereegranskat)abstract
- A picornavirus (Ljungan virus) has been associated with diabetes in its wild rodent reservoir and in diabetesprone biobreeding (DP-BB) rats. We attempted to alter the development of diabetes in DP-BB rats using two anti-picornavirus compounds (pleconaril and APO-N039), singly or in combination. Antiviral therapy was initiated 2 weeks before expected onset of diabetes. Pleconaril or APO-N039 alone did not affect the debut of diabetes. However, animals receiving a combination of both compounds were protected for at least the entire period of treatment (4 weeks after expected time of diabetes onset). Immunohistochemistry demonstrated that the presence and distribution of virus antigen in the pancreatic islets coincided with the clinical status of the animal. Data indicate that a treatable picornavirus can be involved in the cellular assault resulting in diabetes and in these cases the disease mechanism appears to involve a virus present in the pancreatic beta cell mass itself.
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| 4. |
- Tolf, Conny, et al.
(författare)
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Characterization of polyclonal antibodies against the capsid proteins of Ljungan virus
- 2008
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Ingår i: Journal of Virological Methods. - : Elsevier BV. - 0166-0934 .- 1879-0984. ; 150:1-2, s. 34-40
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Tidskriftsartikel (refereegranskat)abstract
- Ljungan virus (LV) is a suspected human pathogen isolated from voles in Sweden and North America. To enable virus detection and studies of localization and activity of virion proteins, polyclonal antibodies were produced against bacterially expressed capsid proteins of the LV strain, 87-012G. Specific detection of proteins corresponding to viral antigens in lysates of LV infected cells was demonstrated by immunoblotting using each one of the generated polyclonal antibodies. In addition, native viral antigens present in cell culture infected with LV strains 87-012G or 145SLG were detected in ELISA and by immunofluorescence using the antibodies against the VP0 and VP1 proteins. The anti-VP3 antibody did not react with native proteins of the LV virion, suggesting that the VP3 is less potent in evoking humoral response and may have a less exposed orientation in the virus capsid. No activity of the antibodies was observed against the closely related human parechovirus type 1. The polyclonal antibody against the VP1 protein was further used for detection of LV infected myocytes in a mouse model of LV-induced myocarditis. Thus, polyclonal antibodies against recombinant viral capsid proteins enabled detection of natural LV virions by several different immunological methods.
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