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Olli Vapalahti - One of the best experts on this subject based on the ideXlab platform.
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vaccinia Virus free rescue of fluorescent replication defective vesicular stomatitis Virus and pseudotyping with Puumala Virus glycoproteins for use in neutralization tests
Journal of General Virology, 2016Co-Authors: Rommel Paneth Iheozorejiofor, Ake Lundkvist, Alexander Plyusnin, Lev Levanov, Jussi Hepojoki, Tomas Strandin, Olli VapalahtiAbstract:Puumala Virus (PUUV) grows slowly in cell culture. To study antigenic properties of PUUV, an amenable method for their expression would be beneficial. To achieve this, a replication-defective recombinant vesicular stomatitis Virus, rVSVΔG*EGFP, was rescued using BSRT7/5 and encephalomyocarditis Virus (EMCV) internal ribosomal entry site (IRES)-enabled rescue plasmids. Using these particles, pseudotypes bearing PUUV Sotkamo strain glycoproteins were produced, with titres in the range 105–108, and were used in pseudotype focus reduction neutralization tests (pFRNTs) with neutralizing monoclonal antibodies and patient sera. The results were compared with those from orthodox focus reduction neutralization tests (oFRNTs) using native PUUV with the same samples and showed a strong positive correlation (rs = 0.82) between the methods. While developing the system we identified three amino acids which were mutated in the Vero E6 cell culture adapted PUUV prototype Sotkamo strain sequence, and changing these residues was critical for expression and neutralizing antibody binding of PUUV glycoproteins.
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serological survey of seewis Virus antibodies in patients suspected for hantaVirus infection in finland a cross reaction between Puumala Virus antiserum with seewis Virus n protein
Journal of General Virology, 2015Co-Authors: Jiaxin Ling, Olli Vapalahti, Heikki Henttonen, Antti Vaheri, Tarja Sironen, Lev Levanov, Satu Hepojoki, Anne J Jaaskelainen, Jussi HepojokiAbstract:Puumala Virus (PUUV, carried by Myodes glareolus) co-circulates with Seewis Virus (SWSV, carried by Sorex araneus) in Finland. While PUUV causes 1000–3000 nephropathia epidemica (NE) cases annually, the pathogenicity of SWSV to man is unknown. To study the prevalence of SWSV antibodies in hantaVirus fever-like patients' sera, we used recombinant SWSV nucleocapsid (N) protein as the antigen in ELISA, immunofluorescence assay (IFA) and immunoblotting. While characterizing the recombinant SWSV N protein, we observed that a polyclonal rabbit antiserum against PUUV N protein cross-reacted with SWSV N protein and vice versa. We initially screened 486 (450 PUUV-seronegative and 36 PUUV-seropositive) samples sent to Helsinki University Hospital Laboratory for PUUV serodiagnosis during 2002 and 2007 in an SWSV N protein IgG ELISA. In total, 4.2 % (19/450) of the PUUV-seronegative samples were reactive in the SWSV N protein IgG ELISA and none of the tested samples [43 PUUV-seronegative (weakly reactive in the SWSV IgG ELISA) and 15 random] were reactive in the SWSV N protein IgM ELISA. None of the IgG reactions could be confirmed by IFA or immunoblotting. Furthermore, among the 36 PUUV-seropositive samples three were reactive in SWSV N protein IgG and ten in SWSV N protein IgM ELISA. One PUUV-seropositive sample reacted with SWSV N protein in IFA and four in immunoblotting. Finally, we applied competitive ELISA to confirm that the observed reactivity was due to cross-reactivity rather than a true SWSV response. In conclusion, no evidence of SWSV infection was found among the 486 samples studied; however, we did demonstrate that PUUV antiserum cross-reacted with shrew-borne hantaVirus N protein.
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Analysis of complete Puumala Virus genome, Finland.
Emerging Infectious Diseases, 2012Co-Authors: Angelina Plyusnina, Olli Vapalahti, Heikki Henttonen, Antti Vaheri, Maria Razzauti, Tarja Sironen, Jukka Niemimaa, Alexander PlyusninAbstract:Puumala Virus causes nephropathia epidemica, a rodent-borne zoonosis that is endemic to Europe. We sequenced the complete Puumala Virus genome that was directly recovered from a person who died and compared it with those of Viruses from local bank voles. The Virus strain involved was neither a unique nor rare genetic variant.
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disease burden of Puumala Virus infections 1995 2008
Epidemiology and Infection, 2010Co-Authors: P Makary, Olli Vapalahti, Mari Kanerva, Jukka Ollgren, M J Virtanen, Outi LyytikainenAbstract:Puumala Virus (PUUV) causes mild haemorrhagic fever with renal syndrome, a rodent-borne zoonosis. To evaluate the disease burden of PUUV infections in Finland, we analysed data reported by laboratories to the National Infectious Disease Registry during 1995―2008 and compared these with data from other national registries (death, 1998―2007; hospital discharge, 1996―2007; occupational diseases, 1995―2006). A total of 22681 cases were reported (average annual incidence 31/100 000 population); 85 % were in persons aged 20―64 years and 62 % were males. There was an increasing trend in incidence, and the rates varied widely by season and region. We observed 13 deaths attributable to PUUV infection (case-fatality proportion 0·08%). Of all cases, 9599 (52 %) were hospitalized. Only 590 cases (3 %) were registered as occupational disease, of which most were related to farming and forestry. The wide seasonal and geographical variation is probably related to rodent density and human behaviour.
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Association between the DQA MHC class II gene and Puumala Virus infection in Myodes glareolus, the bank vole
Infection Genetics and Evolution, 2008Co-Authors: Julie Deter, Olli Vapalahti, Liina Voutilainen, Maxime Galan, Heikki Henttonen, Antti Vaheri, Josef Bryja, Yannick Chaval, Juha Laakkonen, Alexis Ribas SalvadorAbstract:Puumala Virus (PUUV) is a hantaVirus specifically harboured by the bank vole, Myodes (earlier Clethrionomys) glareolus. It causes a mild form of hemorrhagic fever with renal syndrome (HFRS) in humans, called Nephropathia epidemica (NE). The clinical severity of NE is variable among patients and depends on their major histocompatibility complex (MHC) genetic background. In this study we investigated the potential role of class II MHC gene polymorphism in the susceptibility/resistance to PUUV in the wild reservoir M. glareolus. We performed an association study between the exon 2 of the DQA gene and PUUV antibodies considering a natural population of bank voles. Because immune gene polymorphism is likely to be driven by multiple parasites in the wild, we also screened bank voles for other potential viral and parasitic infections. We used multivariate analyses to explore DQA polymorphism/PUUV associations while considering the potential antagonist and/or synergistic effects of the whole parasite community. Our study suggests links between class II MHC characteristics and viral infections including PUUV and Cowpox Virus. Several alleles are likely to be involved in the susceptibility or in the resistance of bank voles to these infections. Alternatively, heterozygosity does not seem to be associated with PUUV or any other parasite infections. This result thus provides no evidence in favour of the hypothesis of selection through overdominance. Finally this multivariate approach reveals a strong antagonism between ectoparasitic mites and PUUV, suggesting direct or indirect immunogenetic links between infections by these parasites. Other datasets are now required to confirm these results and to test whether the associations vary in space and/or time.
Antti Vaheri - One of the best experts on this subject based on the ideXlab platform.
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monocyte subset redistribution from blood to kidneys in patients with Puumala Virus caused hemorrhagic fever with renal syndrome
PLOS Pathogens, 2021Co-Authors: Sindhu Vangeti, Tomas Strandin, Jukka Mustonen, Satu Makela, Sang Liu, Johanna Tauriainen, Anne Raisanensokolowski, Luz E Cabrera, Antti Hassinen, Antti VaheriAbstract:Innate immune cells like monocytes patrol the vasculature and mucosal surfaces, recognize pathogens, rapidly redistribute to affected tissues and cause inflammation by secretion of cytokines. We previously showed that monocytes are reduced in blood but accumulate in the airways of patients with Puumala Virus (PUUV) caused hemorrhagic fever with renal syndrome (HFRS). However, the dynamics of monocyte infiltration to the kidneys during HFRS, and its impact on disease severity are currently unknown. Here, we examined longitudinal peripheral blood samples and renal biopsies from HFRS patients and performed in vitro experiments to investigate the fate of monocytes during HFRS. During the early stages of HFRS, circulating CD14-CD16+ nonclassical monocytes (NCMs) that patrol the vasculature were reduced in most patients. Instead, CD14+CD16- classical (CMs) and CD14+CD16+ intermediate monocytes (IMs) were increased in blood, in particular in HFRS patients with more severe disease. Blood monocytes from patients with acute HFRS expressed higher levels of HLA-DR, the endothelial adhesion marker CD62L and the chemokine receptors CCR7 and CCR2, as compared to convalescence, suggesting monocyte activation and migration to peripheral tissues during acute HFRS. Supporting this hypothesis, increased numbers of HLA-DR+, CD14+, CD16+ and CD68+ cells were observed in the renal tissues of acute HFRS patients compared to controls. In vitro, blood CD16+ monocytes upregulated CD62L after direct exposure to PUUV whereas CD16- monocytes upregulated CCR7 after contact with PUUV-infected endothelial cells, suggesting differential mechanisms of activation and response between monocyte subsets. Together, our findings suggest that NCMs are reduced in blood, potentially via CD62L-mediated attachment to endothelial cells and monocytes are recruited to the kidneys during HFRS. Monocyte mobilization, activation and functional impairment together may influence the severity of disease in acute PUUV-HFRS.
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serological survey of seewis Virus antibodies in patients suspected for hantaVirus infection in finland a cross reaction between Puumala Virus antiserum with seewis Virus n protein
Journal of General Virology, 2015Co-Authors: Jiaxin Ling, Olli Vapalahti, Heikki Henttonen, Antti Vaheri, Tarja Sironen, Lev Levanov, Satu Hepojoki, Anne J Jaaskelainen, Jussi HepojokiAbstract:Puumala Virus (PUUV, carried by Myodes glareolus) co-circulates with Seewis Virus (SWSV, carried by Sorex araneus) in Finland. While PUUV causes 1000–3000 nephropathia epidemica (NE) cases annually, the pathogenicity of SWSV to man is unknown. To study the prevalence of SWSV antibodies in hantaVirus fever-like patients' sera, we used recombinant SWSV nucleocapsid (N) protein as the antigen in ELISA, immunofluorescence assay (IFA) and immunoblotting. While characterizing the recombinant SWSV N protein, we observed that a polyclonal rabbit antiserum against PUUV N protein cross-reacted with SWSV N protein and vice versa. We initially screened 486 (450 PUUV-seronegative and 36 PUUV-seropositive) samples sent to Helsinki University Hospital Laboratory for PUUV serodiagnosis during 2002 and 2007 in an SWSV N protein IgG ELISA. In total, 4.2 % (19/450) of the PUUV-seronegative samples were reactive in the SWSV N protein IgG ELISA and none of the tested samples [43 PUUV-seronegative (weakly reactive in the SWSV IgG ELISA) and 15 random] were reactive in the SWSV N protein IgM ELISA. None of the IgG reactions could be confirmed by IFA or immunoblotting. Furthermore, among the 36 PUUV-seropositive samples three were reactive in SWSV N protein IgG and ten in SWSV N protein IgM ELISA. One PUUV-seropositive sample reacted with SWSV N protein in IFA and four in immunoblotting. Finally, we applied competitive ELISA to confirm that the observed reactivity was due to cross-reactivity rather than a true SWSV response. In conclusion, no evidence of SWSV infection was found among the 486 samples studied; however, we did demonstrate that PUUV antiserum cross-reacted with shrew-borne hantaVirus N protein.
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Analysis of complete Puumala Virus genome, Finland.
Emerging Infectious Diseases, 2012Co-Authors: Angelina Plyusnina, Olli Vapalahti, Heikki Henttonen, Antti Vaheri, Maria Razzauti, Tarja Sironen, Jukka Niemimaa, Alexander PlyusninAbstract:Puumala Virus causes nephropathia epidemica, a rodent-borne zoonosis that is endemic to Europe. We sequenced the complete Puumala Virus genome that was directly recovered from a person who died and compared it with those of Viruses from local bank voles. The Virus strain involved was neither a unique nor rare genetic variant.
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Association between the DQA MHC class II gene and Puumala Virus infection in Myodes glareolus, the bank vole
Infection Genetics and Evolution, 2008Co-Authors: Julie Deter, Olli Vapalahti, Liina Voutilainen, Maxime Galan, Heikki Henttonen, Antti Vaheri, Josef Bryja, Yannick Chaval, Juha Laakkonen, Alexis Ribas SalvadorAbstract:Puumala Virus (PUUV) is a hantaVirus specifically harboured by the bank vole, Myodes (earlier Clethrionomys) glareolus. It causes a mild form of hemorrhagic fever with renal syndrome (HFRS) in humans, called Nephropathia epidemica (NE). The clinical severity of NE is variable among patients and depends on their major histocompatibility complex (MHC) genetic background. In this study we investigated the potential role of class II MHC gene polymorphism in the susceptibility/resistance to PUUV in the wild reservoir M. glareolus. We performed an association study between the exon 2 of the DQA gene and PUUV antibodies considering a natural population of bank voles. Because immune gene polymorphism is likely to be driven by multiple parasites in the wild, we also screened bank voles for other potential viral and parasitic infections. We used multivariate analyses to explore DQA polymorphism/PUUV associations while considering the potential antagonist and/or synergistic effects of the whole parasite community. Our study suggests links between class II MHC characteristics and viral infections including PUUV and Cowpox Virus. Several alleles are likely to be involved in the susceptibility or in the resistance of bank voles to these infections. Alternatively, heterozygosity does not seem to be associated with PUUV or any other parasite infections. This result thus provides no evidence in favour of the hypothesis of selection through overdominance. Finally this multivariate approach reveals a strong antagonism between ectoparasitic mites and PUUV, suggesting direct or indirect immunogenetic links between infections by these parasites. Other datasets are now required to confirm these results and to test whether the associations vary in space and/or time.
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Immune responses to Puumala Virus infection and the pathogenesis of nephropathia epidemica
Microbes and infection, 2004Co-Authors: Masanori Terajima, Olli Vapalahti, Antti Vaheri, Heather L. Van Epps, Francis A. EnnisAbstract:Puumala Virus, causative agent of a mild form of hemorrhagic fever with renal syndrome, also known as nephropathia epidemica, induces long-lasting humoral and cellular immunity in patients. The Virus itself is not cytopathic, and the immune responses to the Virus may be involved in the pathogenesis of the disease.
Ake Lundkvist - One of the best experts on this subject based on the ideXlab platform.
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vaccinia Virus free rescue of fluorescent replication defective vesicular stomatitis Virus and pseudotyping with Puumala Virus glycoproteins for use in neutralization tests
Journal of General Virology, 2016Co-Authors: Rommel Paneth Iheozorejiofor, Ake Lundkvist, Alexander Plyusnin, Lev Levanov, Jussi Hepojoki, Tomas Strandin, Olli VapalahtiAbstract:Puumala Virus (PUUV) grows slowly in cell culture. To study antigenic properties of PUUV, an amenable method for their expression would be beneficial. To achieve this, a replication-defective recombinant vesicular stomatitis Virus, rVSVΔG*EGFP, was rescued using BSRT7/5 and encephalomyocarditis Virus (EMCV) internal ribosomal entry site (IRES)-enabled rescue plasmids. Using these particles, pseudotypes bearing PUUV Sotkamo strain glycoproteins were produced, with titres in the range 105–108, and were used in pseudotype focus reduction neutralization tests (pFRNTs) with neutralizing monoclonal antibodies and patient sera. The results were compared with those from orthodox focus reduction neutralization tests (oFRNTs) using native PUUV with the same samples and showed a strong positive correlation (rs = 0.82) between the methods. While developing the system we identified three amino acids which were mutated in the Vero E6 cell culture adapted PUUV prototype Sotkamo strain sequence, and changing these residues was critical for expression and neutralizing antibody binding of PUUV glycoproteins.
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associations between mhc genes and Puumala Virus infection in myodes glareolus are detected in wild populations but not from experimental infection data
Journal of General Virology, 2010Co-Authors: Emmanuel Guivier, Eva R Kallio, Pierrejean G Male, Liina Voutilainen, Maxime Galan, Ake Lundkvist, Heikki Henttonen, Katrien Tersago, Gert E. Olsson, Denis AugotAbstract:We analysed the influence of MHC class II Dqa and Drb genes on Puumala Virus (PUUV) infection in bank voles (Myodes glareolus). We considered voles sampled in five European localities or derived from a previous experiment that showed variable infection success of PUUV. The genetic variation observed in the Dqa and Drb genes was assessed by using single-strand conformation polymorphism and pyrosequencing methods, respectively. Patterns were compared with those obtained from 13 microsatellites. We revealed significant genetic differentiation between PUUV-seronegative and -seropositive bank voles sampled in wild populations, at the Drb gene only. The absence of genetic differentiation observed at neutral microsatellites confirmed the important role of selective pressures in shaping these Drb patterns. Also, we found no significant associations between infection success and MHC alleles among laboratory-colonized bank voles, which is explained by a loss of genetic variability that occurred during the captivity of these voles.
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associations between mhc genes and Puumala Virus infection in myodes glareolus are detected in wild populations but not from experimental infection data
Journal of General Virology, 2010Co-Authors: Emmanuel Guivier, Eva R Kallio, Pierrejean G Male, Liina Voutilainen, Maxime Galan, Ake Lundkvist, Heikki Henttonen, Katrien Tersago, Gert E. Olsson, Denis AugotAbstract:We analysed the influence of MHC class II Dqa and Drb genes on Puumala Virus (PUUV) infection in bank voles (Myodes glareolus). We considered voles sampled in five European localities or derived from a previous experiment that showed variable infection success of PUUV. The genetic variation observed in the Dqa and Drb genes was assessed by using single-strand conformation polymorphism and pyrosequencing methods, respectively. Patterns were compared with those obtained from 13 microsatellites. We revealed significant genetic differentiation between PUUV-seronegative and -seropositive bank voles sampled in wild populations, at the Drb gene only. The absence of genetic differentiation observed at neutral microsatellites confirmed the important role of selective pressures in shaping these Drb patterns. Also, we found no significant associations between infection success and MHC alleles among laboratory-colonized bank voles, which is explained by a loss of genetic variability that occurred during the captivity of these voles.
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Improvement of binding of Puumala Virus neutralization site resembling peptide with a second-generation phage library
Protein Engineering, 2003Co-Authors: Tuomas Heiskanen, Ake Lundkvist, Antti Vaheri, Jussi Hepojoki, Xiao Dong Li, Elisabeth Gustafsson, Hilkka LankinenAbstract:We have previously selected a peptide insert FPCDRLSGYWERGIPSPCVR recognizing the Puumala Virus (PUUV) G2-glycoprotein-specific neutralizing monoclonal antibody (MAb) 1C9 with K d of 2.85×10 - 8 from a random peptide library X 2 CX 1 4 CX 2 expressed on the pIII protein of the filamentous phage fd-tet. We have now created a second-generation phage-displayed peptide library in which each amino acid of the peptide was mutated randomly to another with a certain probability. Peptides were selected for higher affinity for MAb 1C9 and for a common binding motif for MAb 4G2 having an overlapping epitope with MAb 1C9 in G2 glycoprotein. The resulting peptides were synthesized as spots on cellulose membrane. Amino acid changes which improved the reactivity of the peptides to MAb 1C9 were combined in the peptide ATCDKLFGYYERGIPLPCAL with K d of 1.49×10 - 9 in biosensor measurements. Our results show that the binding properties of peptides, the affinity and the specificity can be improved and the binding specificity determining amino acids and structural factors can be analyzed by combining binding assays with synthetic peptides on membrane with the use of second-generation phage display libraries.
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new immunochromatographic rapid test for diagnosis of acute Puumala Virus infection
Journal of Clinical Microbiology, 2001Co-Authors: Helena Hujakka, Ake Lundkvist, Antti Vaheri, Vesa Koistinen, Pekka Eerikainen, Ilpo Kuronen, Ilkka Mononen, Markku Parviainen, Ale Narvanen, Olli VapalahtiAbstract:A new immunochromatographic rapid test, POC Puumala (Erilab Ltd., Kuopio, Finland), for detection of acute-phase Puumala Virus (PUUV) infection was developed based on a highly purified baculoVirus-expressed PUUV nucleocapsid protein antigen and lateral immunodiffusion techniques. After addition of sample (5 μl of serum, plasma, or fingertip blood) and buffer, PUUV-specific immunoglobulin M (IgM) antibodies, if present, together with the gold-conjugated anti-human IgM, formed a specific colored line in 5 min. The sensitivity and specificity of the test were evaluated with 200 serum samples and 30 fingertip blood samples. The reference method for the serum samples was a μ-capture enzyme immunoassay (EIA) for IgM and an immunofluorescence assay (IFA) for IgG antibodies. The analytical sensitivity and specificity of the rapid test were 100 and 99%, respectively, for unfrozen serum samples (n = 103; 12 PUUV IgM-positive samples). When freeze-thawed serum samples were used, the sensitivity and specificity were each 97.1% (n = 70; 35 PUUV IgM-positive samples). The specificity of the test was 96.2% for 27 serum samples with nonspecific IgM antibodies or rheumatoid factor (RF). The fingertip blood samples (n = 30) were negative, but they gave clear positive results when spiked with IgM-positive sera (n = 20). The results were in good agreement with the standard diagnostic methods. The rapid performance, the lack of need for refined laboratory equipment, and the high specificity with fresh serum and fingertip blood samples indicate that the developed POC Puumala rapid test is a useful tool for fast diagnosis of acute PUUV infection.
Rainer G Ulrich - One of the best experts on this subject based on the ideXlab platform.
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A highly divergent Puumala Virus lineage in southern Poland
Archives of Virology, 2017Co-Authors: Ulrike M. Rosenfeld, Stephan Drewes, Gerald Heckel, Edyta T. Sadowska, Magdalena Mikowska, Paweł Koteja, Rainer G UlrichAbstract:Puumala Virus (PUUV) represents one of the most important hantaViruses in Central Europe. Phylogenetic analyses of PUUV strains indicate a strong genetic structuring of this hantaVirus. Recently, PUUV sequences were identified in the natural reservoir, the bank vole ( Myodes glareolus ), collected in the northern part of Poland. The objective of this study was to evaluate the presence of PUUV in bank voles from southern Poland. A total of 72 bank voles were trapped in 2009 at six sites in this part of Poland. RT-PCR and IgG-ELISA analyses detected three PUUV positive voles at one trapping site. The PUUV-infected animals were identified by cytochrome b gene analysis to belong to the Carpathian and Eastern evolutionary lineages of bank vole. The novel PUUV S, M and L segment nucleotide sequences showed the closest similarity to sequences of the Russian PUUV lineage from Latvia, but were highly divergent to those previously found in northern Poland, Slovakia and Austria. In conclusion, the detection of a highly divergent PUUV lineage in southern Poland indicates the necessity of further bank vole monitoring in this region allowing rational public health measures to prevent human infections.
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Puumala Virus in bank voles lithuania
Emerging Infectious Diseases, 2017Co-Authors: Petra Strakova, Sandra Jagdmann, Linas Balciauskas, Laima Balciauskienė, Stephan Drewes, Rainer G UlrichAbstract:Little is known about the presence of human pathogenic Puumala Virus (PUUV) in Lithuania. We detected this Virus in bank voles (Myodes glareolus) in a region of this country in which previously PUUV-seropositive humans were identified. Our results are consistent with heterogeneous distributions of PUUV in other countries in Europe.
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host associated absence of human Puumala Virus infections in northern and eastern germany
Emerging Infectious Diseases, 2017Co-Authors: Stephan Drewes, Ulrike Rosenfeld, Hanan Sheikh Ali, Mathias Schlegel, Moritz Saxenhofer, Florian Binder, Fabian Cuypers, Susanne Rohrs, Gerald Heckel, Rainer G UlrichAbstract:Human hantaVirus disease cases, caused by Puumala Virus (PUUV), are mainly recorded in western and southern areas of Germany. This bank vole reservoir survey confirmed PUUV presence in these regions but its absence in northern and eastern regions. PUUV occurrence is associated with the presence of the Western bank vole phylogroup.
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complete genome of a Puumala Virus strain from central europe
Virus Genes, 2015Co-Authors: Hanan Sheikh Ali, Mathias Schlegel, Stephan Drewes, Gerald Heckel, Vanessa Weber De Melo, J Freise, Martin H Groschup, Rainer G UlrichAbstract:Puumala Virus (PUUV) is one of the predominant hantaVirus species in Europe causing mild to moderate cases of haemorrhagic fever with renal syndrome. Parts of Lower Saxony in north-western Germany are endemic for PUUV infections. In this study, the complete PUUV genome sequence of a bank vole-derived tissue sample from the 2007 outbreak was determined by a combined primer-walking and RNA ligation strategy. The S, M and L genome segments were 1,828, 3,680 and 6,550 nucleotides in length, respectively. Sliding-window analyses of the nucleotide sequences of all available complete PUUV genomes indicated a non-homogenous distribution of variability with hypervariable regions located at the 3′-ends of the S and M segments. The overall similarity of the coding genome regions to the other PUUV strains ranged between 80.1 and 84.7 % at the level of the nucleotide sequence and between 89.5 and 98.1 % for the deduced amino acid sequences. In comparison to the phylogenetic trees of the complete coding sequences, trees based on partial segments revealed a general drop in phylogenetic support and a lower resolution. The Astrup strain S and M segment sequences showed the highest similarity to sequences of strains from geographically close sites in the Osnabruck Hills region. In conclusion, a primer-walking-mediated strategy resulted in the determination of the first complete nucleotide sequence of a PUUV strain from Central Europe. Different levels of variability along the genome provide the opportunity to choose regions for analyses according to the particular research question, e.g., large-scale phylogenetics or within-host evolution.
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Puumala Virus outbreak in western thuringia germany 2010 epidemiology and strain identification
Zoonoses and Public Health, 2013Co-Authors: Mirko Faber, Mathias Schlegel, Rainer G Ulrich, T Wollny, Konrad M Wanka, Jorg Thiel, Christina Frank, D Rimek, Klaus StarkAbstract:In 2010, the highest annual number of human Puumala Virus (PUUV) infections was reported in Germany since hantaVirus surveillance started in 2001. The increase in annual case numbers was especially marked in western Thuringia. We combined results of case-based hantaVirus surveillance in humans and serological and molecular investigations in the rodent reservoir to describe the epidemiological situation and to identify the putative outbreak strain. A 5-fold increase in notified hantaVirus cases compared to the previous annual maximum was observed in western Thuringia in 2010. Disease incidence varied tremendously within a small geographical area with case patients' places of residence clustering around beech-dominated broad leaf forest patches. Investigations in the rodent reservoir revealed a novel Puumala Virus (PUUV) subtype, which is clearly distinct from strains collected in other PUUV endemic regions of Germany. It can be assumed that in regions in western Thuringia where hantaVirus cases occurred in 2010 or previous outbreak years, PUUV has been present in the environment for a long time. Further studies are needed to elucidate the population dynamics and hantaVirus prevalence of the rodent reservoir and driving ecological factors.
Alexander Plyusnin - One of the best experts on this subject based on the ideXlab platform.
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vaccinia Virus free rescue of fluorescent replication defective vesicular stomatitis Virus and pseudotyping with Puumala Virus glycoproteins for use in neutralization tests
Journal of General Virology, 2016Co-Authors: Rommel Paneth Iheozorejiofor, Ake Lundkvist, Alexander Plyusnin, Lev Levanov, Jussi Hepojoki, Tomas Strandin, Olli VapalahtiAbstract:Puumala Virus (PUUV) grows slowly in cell culture. To study antigenic properties of PUUV, an amenable method for their expression would be beneficial. To achieve this, a replication-defective recombinant vesicular stomatitis Virus, rVSVΔG*EGFP, was rescued using BSRT7/5 and encephalomyocarditis Virus (EMCV) internal ribosomal entry site (IRES)-enabled rescue plasmids. Using these particles, pseudotypes bearing PUUV Sotkamo strain glycoproteins were produced, with titres in the range 105–108, and were used in pseudotype focus reduction neutralization tests (pFRNTs) with neutralizing monoclonal antibodies and patient sera. The results were compared with those from orthodox focus reduction neutralization tests (oFRNTs) using native PUUV with the same samples and showed a strong positive correlation (rs = 0.82) between the methods. While developing the system we identified three amino acids which were mutated in the Vero E6 cell culture adapted PUUV prototype Sotkamo strain sequence, and changing these residues was critical for expression and neutralizing antibody binding of PUUV glycoproteins.
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Analysis of complete Puumala Virus genome, Finland.
Emerging Infectious Diseases, 2012Co-Authors: Angelina Plyusnina, Olli Vapalahti, Heikki Henttonen, Antti Vaheri, Maria Razzauti, Tarja Sironen, Jukka Niemimaa, Alexander PlyusninAbstract:Puumala Virus causes nephropathia epidemica, a rodent-borne zoonosis that is endemic to Europe. We sequenced the complete Puumala Virus genome that was directly recovered from a person who died and compared it with those of Viruses from local bank voles. The Virus strain involved was neither a unique nor rare genetic variant.
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hypophyseal hemorrhage and panhypopituitarism during Puumala Virus infection magnetic resonance imaging and detection of viral antigen in the hypophysis
Clinical Infectious Diseases, 2002Co-Authors: Timo Hautala, Olli Vapalahti, Antti Vaheri, Tarja Sironen, Alexander Plyusnin, Eija Paakko, Terttu Sarkioja, P I Salmela, Heikki KaumaAbstract:We describe 3 cases of nephropathia epidemica (NE) that confirm that Puumala Virus infection may cause hypophyseal injury. Autopsy revealed a hemorrhagic hypophysis positive for Puumala Virus antigen in both neuroendocrine stromal and vascular endothelial cells in 1 patient, and 2 patients developed hypophyseal hemorrhage (diagnosed with magnetic resonance imaging) during or shortly after acute NE, both of whom developed panhypopituitarism.
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nucleotide and deduced amino acid sequences of the m and s genome segments of a swedish Puumala Virus isolate
Virus Research, 1995Co-Authors: Jan Hörling, Antti Vaheri, Alexander Plyusnin, Bo Niklasson, Heikki Lehvaslaiho, Katarina Persson, Ying Cheng, Ake LundkvistAbstract:Abstract The Swedish Puumala (PUU) Virus strain Vindeln 83-L20, isolated from a bank vole trapped in 1983 near Vindeln, Vasterbotten country, Sweden, was characterized by nucleotide sequence analysis. The coding region of the M segment was determined by PCR followed by direct sequencing and the entire S segment was characterized by cloning and nucleotide sequence analysis. The genomic organization was found to be very similar to that of other PUU Virus strains regarding open reading frames, polypeptide sizes and potential glycosylation sites. According to phylogenetic analysis 83-L20 was found to represent a new lineage within the Puumala Virus serotype in the HantaVirus genus. The M segment sequence of 83-L20 was found to be more closely related to the Finnish PUU Virus strains than to strains from Central Europe or from the Russia. The evolutionary origin of the S segment was not as clearly resolved since the branching points of all PUU Virus strains in the phylogenetic tree were nearly the same.
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human b cell epitopes of Puumala Virus nucleocapsid protein the major antigen in early serological response
Journal of Medical Virology, 1995Co-Authors: Olli Vapalahti, Ake Lundkvist, Antti Vaheri, Alexander Plyusnin, Hannimari Kalliokokko, M Brummerkorvenkontio, Heikki Lehvaslaiho, Ale Narvanen, Ilkka Julkunen, Hilkka LankinenAbstract:Puumala Virus (PUU) is a member of the Hantavi rus genus in the family Bunyaviridae and the etiologic agent of nephropathia epidemica (NE), a form of haemorrhagic fever with renal syndrome (HFRS). In this study we compared the immunofluorescence patterns of NE sera and antibodies raised against recombinant PUU proteins and confirm that the nucleocapsid protein is the major target in the early IgG response of NE patients and provides the molecular basis for simple and rapid differentiation between acute illness and old immunity by granular vs. diffuse fluorescence staining in the indirect immunofluorescence test. The differential kinetics of B-cell responses to PUU nucleocapsid vs. envelope proteins was emphasized further by the end-point titres of IgG antibodies to N, G1 and G2 proteins in NE patients. The granular fluorescence correlated with low IgG avidity in 99.8%. and diffuse fluorescence with high avidity in 100% of 617 NE sera studied. Epitope scanning with overlapping 14-mer peptides covering the whole nucleocapsid protein by a shift of 3 amino acids revealed six major antigenic epitopes recognized by sera from acute-phase NE patients. The epitopes clustered mainly in the hydrophilic regions, and two of them in a highly variable region which could probably serve as an antigen to distinguish serologically between infections of closely related hantaViruses, some apparently apathogenic, some causing lethal infections. The anti-peptide epitope pattern varied between different individuals and a collection of several pinbound peptides was needed to be recognised by most NE sera studied. © 1995 Wiley-Liss, Inc.
