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Gullberg, Maria
Publications (10 of 12) Show all publications
Jonsson, N., Sävneby, A., Gullberg, M., Evertsson, K., Klingel, K. & Lindberg, A. M. (2015). Efficient replication of recombinant Enterovirus B types, carrying different P1 genes in the coxsackievirus B5 replicative backbone. Virus genes, 50(3), 351-357
Open this publication in new window or tab >>Efficient replication of recombinant Enterovirus B types, carrying different P1 genes in the coxsackievirus B5 replicative backbone
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2015 (English)In: Virus genes, ISSN 0920-8569, E-ISSN 1572-994X, Vol. 50, no 3, p. 351-357Article in journal (Refereed) Published
Abstract [en]

Recombination is an important feature in theevolution of the Enterovirus genus. Phylogenetic studies ofenteroviruses have revealed that the capsid genomic region(P1) is type specific, while the parts of the genome codingfor the non-structural proteins (P2–P3) are species specific.Hence, the genome may be regarded as consisting of twomodules that evolve independently. In this study, it wasinvestigated whether the non-structural coding part of thegenome in one type could support replication of a virus witha P1 region from another type of the same species. A cas-sette vector (pCas) containing a full-length cDNA copy ofcoxsackievirus B5 (CVB5) was used as a replicative back-bone. The P1 region of pCas was replaced with the corre-sponding part from coxsackievirus B3Nancy(CVB3N),coxsackievirus B6Schmitt(CVB6S), and echovirus 7Wal-lace(E7W), all members of theEnterovirus Bspecies. Thereplication efficiency after transfection with clone-derivedin vitro transcribed RNA was studied and compared withthat of pCas. All the recombinant viruses replicated with similar efficiencies and showed threshold cycle (Ct) values,tissue culture infectivity dose 50 %, and plaque-forming unittiters comparable to viruses generated from the pCas con-struct. In addition to this, a clone without the P1 region wasalso constructed, and Western Blot and immunofluorescencestaining analysis showed that the viral genome could betranslated and replicated despite the lack of the structuralprotein-coding region. To conclude, the replicative back-bone of the CVB5 cassette vector supports replication ofintraspecies constructs with P1 regions derived from othermembers of theEnterovirus Bspecies. In addition to this,the replicative backbone can be both translated and repli-cated without the presence of a P1 region.

National Category
Biochemistry and Molecular Biology
Research subject
Biomedical Sciences, Virology
Identifiers
urn:nbn:se:lnu:diva-41538 (URN)10.1007/s11262-015-1177-x (DOI)000355233000001 ()25663145 (PubMedID)2-s2.0-84929956418 (Scopus ID)
Available from: 2015-04-01 Created: 2015-04-01 Last updated: 2017-12-04Bibliographically approved
Persson, M., Gullberg, M., Tolf, C., Lindberg, A. M., Månsson, A. & Kocer, A. (2013). Transportation of Nanoscale Cargoes by Myosin Propelled Actin Filaments. PLoS ONE, 8(2), Article ID e55931.
Open this publication in new window or tab >>Transportation of Nanoscale Cargoes by Myosin Propelled Actin Filaments
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2013 (English)In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 8, no 2, article id e55931Article in journal (Refereed) Published
Abstract [en]

Myosin II propelled actin filaments move ten times faster than kinesin driven microtubules and are thus attractive candidates as cargo-transporting shuttles in motor driven lab-on-a-chip devices. In addition, actomyosin-based transportation of nanoparticles is useful in various fundamental studies. However, it is poorly understood how actomyosin function is affected by different number of nanoscale cargoes, by cargo size, and by the mode of cargo-attachment to the actin filament. This is studied here using biotin/fluorophores, streptavidin, streptavidin-coated quantum dots, and liposomes as model cargoes attached to monomers along the actin filaments ("side-attached") or to the trailing filament end via the plus end capping protein CapZ. Long-distance transportation (> 100 mu m) could be seen for all cargoes independently of attachment mode but the fraction of motile filaments decreased with increasing number of side-attached cargoes, a reduction that occurred within a range of 10-50 streptavidin molecules, 1-10 quantum dots or with just 1 liposome. However, as observed by monitoring these motile filaments with the attached cargo, the velocity was little affected. This also applied for end-attached cargoes where the attachment was mediated by CapZ. The results with side-attached cargoes argue against certain models for chemomechanical energy transduction in actomyosin and give important insights of relevance for effective exploitation of actomyosin-based cargo-transportation in molecular diagnostics and other nanotechnological applications. The attachment of quantum dots via CapZ, without appreciable modulation of actomyosin function, is useful in fundamental studies as exemplified here by tracking with nanometer accuracy.

National Category
Biochemistry and Molecular Biology
Research subject
Natural Science, Biomedical Sciences
Identifiers
urn:nbn:se:lnu:diva-24860 (URN)10.1371/journal.pone.0055931 (DOI)000315186000012 ()2-s2.0-84874337312 (Scopus ID)
Available from: 2013-03-22 Created: 2013-03-22 Last updated: 2017-12-06Bibliographically approved
EL Hiar, R., Haddad, S., Jaidane, H., Hober, D., Ben M'hadheb-Gharbi, M., Gullberg, M., . . . Aouni, M. (2012). Enteroviral Central Nervous System Infections in Children of the Region of Monastir, Tunisia: Diagnosis, Laboratory Findings of Cerebrospinal Fluid and Clinical Manifestations. Indian Journal of Virology, 23(3), 294-302
Open this publication in new window or tab >>Enteroviral Central Nervous System Infections in Children of the Region of Monastir, Tunisia: Diagnosis, Laboratory Findings of Cerebrospinal Fluid and Clinical Manifestations
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2012 (English)In: Indian Journal of Virology, ISSN 0970-2822, Vol. 23, no 3, p. 294-302Article in journal (Refereed) Published
Abstract [en]

Human enteroviruses (HEV) are one of the major causes of central nervous system (CNS) infections in pediatrics. A prospective study was conducted to assess the epidemiological, clinical, and laboratory characteristics of enterovirus (EV) infections of the CNS in children under 15-years-old, suspected of having viral CNS infections and admitted to the Pediatric Department of Monastir University Hospital, Tunisia. Enteroviral RNA was detected by 5' NCR nested RT-PCR assay in 33 % (20 out of 60) of cerebrospinal fluid specimens, whereas only six samples (10 %) were EV positive in cell culture. EV-positive patients were clustered according to their clinical manifestations, predominantly diagnosed as aseptic meningitis (65 %) and meningoencephalitis (20 %). Fever, headache, vomiting, and neck stiffness were the most pronounced symptoms. Pleocytosis with the predominance of lymphocytes was observed in 60 % of EV positive specimens. Although patients suffering from EV infections were encountered throughout the year, most occurred during spring and summer months. Using VP1-2A nested RT-PCR and sequence analysis, three of the 20 positive HEV were identified as Echovirus (E)-9. This is the first report of a cluster of aseptic meningitis cases caused by E-9 in Monastir.

Keywords
Enterovirus, Cerebrospinal fluid, Children, Molecular typing, Epidemiology
National Category
Microbiology
Research subject
Biomedical Sciences, Virology
Identifiers
urn:nbn:se:lnu:diva-23373 (URN)10.1007/s13337-012-0104-1 (DOI)000312076000006 ()2-s2.0-84879555697 (Scopus ID)
Available from: 2013-01-09 Created: 2013-01-09 Last updated: 2016-10-05Bibliographically approved
Israelsson, S., Jonsson, N., Gullberg, M. & Lindberg, A. M. (2011). Cytolytic replication of echoviruses in colon cancer cell lines. Virology Journal, 8, Article ID e473.
Open this publication in new window or tab >>Cytolytic replication of echoviruses in colon cancer cell lines
2011 (English)In: Virology Journal, ISSN 1743-422X, E-ISSN 1743-422X, Vol. 8, article id e473Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Colorectal cancer is one of the most common cancers in the world, killing nearly 50% of patients afflicted. Though progress is being made within surgery and other complementary treatments, there is still need for new and more effective treatments. Oncolytic virotherapy, meaning that a cancer is cured by viral infection, is a promising field for finding new and improved treatments. We have investigated the oncolytic potential of several low-pathogenic echoviruses with rare clinical occurrence. Echoviruses are members of the enterovirus genus within the family Picornaviridae.

METHODS: Six colon cancer cell lines (CaCo-2, HT29, LoVo, SW480, SW620 and T84) were infected by the human enterovirus B species echovirus 12, 15, 17, 26 and 29, and cytopathic effects as well as viral replication efficacy were investigated. Infectivity was also tested in spheroids grown from HT29 cells.

RESULTS: Echovirus 12, 17, 26 and 29 replicated efficiently in almost all cell lines and were considered highly cytolytic. The infectivity of these four viruses was further evaluated in artificial tumors (spheroids), where it was found that echovirus 12, 17 and 26 easily infected the spheroids.

CONCLUSIONS: We have found that echovirus 12, 17 and 26 have potential as oncolytic agents against colon cancer, by comparing the cytolytic capacity of five low-pathogenic echoviruses in six colon cancer cell lines and in artificial tumors.

National Category
Microbiology
Research subject
Natural Science, Biomedical Sciences
Identifiers
urn:nbn:se:lnu:diva-16513 (URN)10.1186/1743-422X-8-473 (DOI)21999585 (PubMedID)2-s2.0-80054008339 (Scopus ID)
Available from: 2012-01-03 Created: 2012-01-03 Last updated: 2017-12-08Bibliographically approved
Gullberg, M., Tolf, C., Jonsson, N., Polacek, C., Precechtelova, J., Badurova, M., . . . Lindberg, A. M. (2010). A single coxsackievirus B2 capsid residue controls cytolysis and apoptosis in rhabdomyosarcoma cells.. Journal of Virology, 84(12), 5868-5879
Open this publication in new window or tab >>A single coxsackievirus B2 capsid residue controls cytolysis and apoptosis in rhabdomyosarcoma cells.
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2010 (English)In: Journal of Virology, ISSN 0022-538X, E-ISSN 1098-5514, Vol. 84, no 12, p. 5868-5879Article in journal (Refereed) Published
Abstract [en]

Coxsackievirus B2 (CVB2), one of six human pathogens of the group B coxsackieviruses within the enterovirus genus of Picornaviridae, causes a wide spectrum of human diseases ranging from mild upper respiratory illnesses to myocarditis and meningitis. The CVB2 prototype strain Ohio-1 (CVB2O) was originally isolated from a patient with summer grippe in the 1950s. Later on, CVB2O was adapted to cytolytic replication in rhabdomyosarcoma (RD) cells. Here, we present analyses of the correlation between the adaptive mutations of this RD variant and the cytolytic infection in RD cells. Using reverse genetics, we identified a single amino acid change within the exposed region of the VP1 protein (glutamine to lysine at position 164) as the determinant for the acquired cytolytic trait. Moreover, this cytolytic virus induced apoptosis, including caspase activation and DNA degradation, in RD cells. These findings contribute to our understanding of the host cell adaptation process of CVB2O and provide a valuable tool for further studies of virus-host interactions.

National Category
Microbiology in the medical area
Research subject
Biomedical Sciences, Virology
Identifiers
urn:nbn:se:lnu:diva-7168 (URN)10.1128/JVI.02383-09 (DOI)20375176 (PubMedID)2-s2.0-77952716030 (Scopus ID)
Available from: 2010-08-13 Created: 2010-08-13 Last updated: 2020-06-05Bibliographically approved
Gullberg, M., Tolf, C., Jonsson, N., Mulders, M. N., Savolainen-Kopra, C., Hovi, T., . . . Lindberg, A. M. (2010). Characterization of a putative ancestor of coxsackievirus B5.. Journal of Virology, 84, 9695-9708
Open this publication in new window or tab >>Characterization of a putative ancestor of coxsackievirus B5.
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2010 (English)In: Journal of Virology, ISSN 0022-538X, E-ISSN 1098-5514, Vol. 84, p. 9695-9708Article in journal (Refereed) Published
Abstract [en]

Like other RNA viruses, coxsackievirus B5 (CVB5) exists as circulating heterogeneous populations of genetic variants. In this study, we present the reconstruction and characterization of a probable ancestral virion of CVB5. Phylogenetic analyses based on capsid protein encoding regions (the VP1 gene of 41 clinical isolates and the entire P1 region of eight clinical isolates) of CVB5 revealed two major co-circulating lineages. Ancestral capsid sequences were inferred from sequences of these contemporary CVB5 isolates using maximum likelihood methods. By using Bayesian phylodynamic analysis, the inferred VP1 ancestral sequence was dated back to 1854 (1807-1898). In order to study the properties of the putative ancestral capsid, the entire ancestral P1 sequence was synthesized de novo and inserted into the replicative backbone of an infectious CVB5 cDNA clone. Characterization of the recombinant virus in cell culture showed that fully functional infectious virus particles were assembled and that these viruses displayed properties similar to those of modern isolates, in terms of receptor preferences, plaque phenotype, growth characteristics and cell tropism. This is the first report describing resurrection and characterization of a picornavirus with a putative ancestral capsid. Our approach, including phylogenetics-based reconstruction of viral predecessors, could serve as a starting point for experimental studies of viral evolution and might also provide an alternative strategy in the development of vaccines.

National Category
Microbiology in the medical area
Research subject
Biomedical Sciences, Virology
Identifiers
urn:nbn:se:lnu:diva-7170 (URN)10.1128/JVI.00071-10 (DOI)20631132 (PubMedID)2-s2.0-77956827656 (Scopus ID)
Available from: 2010-08-13 Created: 2010-08-13 Last updated: 2018-01-12Bibliographically approved
Israelsson, S., Gullberg, M., Jonsson, N., Roivainen, M., Edman, K. & Lindberg, A. M. (2010). Studies of Echovirus 5 interactions with the cell surface: Heparan sulfate mediates attachment to the host cell.. Virus Research, 151(2), 170-176
Open this publication in new window or tab >>Studies of Echovirus 5 interactions with the cell surface: Heparan sulfate mediates attachment to the host cell.
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2010 (English)In: Virus Research, ISSN 0168-1702, E-ISSN 1872-7492, Vol. 151, no 2, p. 170-176Article in journal (Refereed) Published
Abstract [en]

Infections caused by Echovirus 5 (E5), an enterovirus of the Picornaviridae family, have been associated with fever, rashes and sporadic cases of aseptic meningitis. To elucidate the receptor usage of this virus, the significance of a previously proposed integrin binding arginine-glycine-aspartic acid (RGD) motif found in the VP3 capsid protein was investigated, as well as the capacity of E5 to interact with heparan sulfate on the cell surface. Using the prototype strain E5 Noyce (E5N), an E5N mutant where the aspartic acid of the RGD motif has been substituted to a glutamic acid and clinical E5 isolates, the RGD motif of VP3 was found to be non-essential and hence not involved in integrin receptor binding. However, E5N and clinical E5 isolates interact with heparan sulfate at the cell surface, as demonstrated by virus replication inhibition assays using heparin and heparinase III, and studies of E5 interactions at the cell surface measured by real-time PCR analysis. In conclusion, E5 utilizes heparan sulfate as a cellular receptor, but the RGD motif of VP3 is not essential for E5 infectivity.

National Category
Microbiology in the medical area
Research subject
Biomedical Sciences, Virology
Identifiers
urn:nbn:se:lnu:diva-7169 (URN)10.1016/j.virusres.2010.05.001 (DOI)20466025 (PubMedID)2-s2.0-77954219093 (Scopus ID)
Available from: 2010-08-13 Created: 2010-08-13 Last updated: 2018-01-12Bibliographically approved
Jonsson, N., Gullberg, M., Israelsson, S. & Lindberg, A. M. (2009). A rapid and efficient method for studies of virus interaction at the host cell surface using enteroviruses and real-time PCR. Virology Journal, 6(Article ID: 217)
Open this publication in new window or tab >>A rapid and efficient method for studies of virus interaction at the host cell surface using enteroviruses and real-time PCR
2009 (English)In: Virology Journal, ISSN 1743-422X, E-ISSN 1743-422X, Vol. 6, no Article ID: 217Article in journal (Refereed) Published
Abstract [en]

Background: Measuring virus attachment to host cells is of great importance when trying to identify novel receptors. The presence of a usable receptor is a major determinant of viral host range and cell tropism. Furthermore, identification of appropriate receptors is central for the understanding of viral pathogenesis and gives possibilities to develop antiviral drugs. Attachment is presently measured using radiolabeled and subsequently gradient purified viruses. Traditional methods are expensive and time-consuming and not all viruses are stable during a purification procedure; hence there is room for improvement. Real-time PCR (RT-PCR) has become the standard method to detect and quantify virus infections, including enteroviruses, in clinical samples. For instance, primers directed to the highly conserved 5' untranslated region (5'UTR) of the enterovirus genome enable detection of a wide spectrum of enteroviruses. Here, we evaluate the capacity of the RT-PCR technology to study enterovirus host cell interactions at the cell surface and compare this novel implementation with an established assay using radiolabeled viruses. Results: Both purified and crude viral extracts of CVB5 generated comparable results in attachment studies when analyzed with RT-PCR. In addition, receptor binding studies regarding viruses with coxsackie- nd adenovirus receptor (CAR) and/or decay accelerating factor (DAF) affinity, further demonstrated the possibility to use RT-PCR to measure virus attachment to host cells. Furthermore, the RT-PCR technology and crude viral extracts was used to study attachment with low multiplicity of infection (0.05 x 10(-4)TCID(50)/cell) and low cell numbers (250), which implies the range of potential implementations of the presented technique. Conclusion: We have implemented the well-established RT-PCR technique to measure viral attachment to host cells with high accuracy and reproducibility, at low cost and with less effort than traditional methods. Furthermore, replacing traditional methods with RT-PCR offers the opportunity to use crude virus containing extracts to investigate attachment, which could be considered as a step towards viral attachment studies in a more natural state.

National Category
Microbiology
Research subject
Biomedical Sciences, Virology; Natural Science, Microbiology; Natural Science, Biomedical Sciences
Identifiers
urn:nbn:se:lnu:diva-2142 (URN)10.1186/1743-422X-6-217 (DOI)000273070600001 ()
Available from: 2010-04-06 Created: 2010-04-06 Last updated: 2017-12-12Bibliographically approved
Tolf, C., Gullberg, M., Ekström, J.-O., Jonsson, N. & Lindberg, A. M. (2009). Identification of Ljungan virus VP0 and VP1 amino acid residues associated with cytolytic replication in cultured cells. Archives of Virology, 154(8), 1271-1284
Open this publication in new window or tab >>Identification of Ljungan virus VP0 and VP1 amino acid residues associated with cytolytic replication in cultured cells
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2009 (English)In: Archives of Virology, ISSN 0304-8608, E-ISSN 1432-8798, Vol. 154, no 8, p. 1271-1284Article in journal (Refereed) Published
Abstract [en]

Ljungan virus is a picornavirus isolated from Swedish and North American rodents. Replication of Ljungan virus in cultured cells normally induces a weak and delayed cytopathic effect compared to that of many other picornaviruses. However, efficiently replicating Ljungan virus variants may evolve during serial passages in cell culture. In this study, we evaluate the significance of three substitutions in capsid protein VP0 and VP1 of a cell-culture-adapted variant of the Swedish Ljungan virus 145SL strain. In contrast to the parental strain, this 145SLG variant grows to high titers in green monkey kidney cells and induces a distinct cytopathic effect. Reverse genetic analyses demonstrated that each one of the individual capsid substitutions contributes to lytic replication in cell culture, but also that expression of all three substitutions results in a 100- to 500-fold increase in viral titers compared to viruses encoding single capsid substitutions. In addition, as indicated by detection of activated caspase-3 and DNA fragmentation, there seems to be an association between increased replication efficiency of lytic Ljungan virus variants and induction of an apoptotic response in infected green monkey kidney cells.

National Category
Microbiology
Research subject
Biomedical Sciences, Virology; Natural Science, Microbiology; Natural Science, Biomedical Sciences
Identifiers
urn:nbn:se:lnu:diva-2031 (URN)10.1007/s00705-009-0417-6 (DOI)
Available from: 2010-04-06 Created: 2010-04-06 Last updated: 2017-12-12Bibliographically approved
Tolf, C., Gullberg, M., Johansson, E. S., Tesh, R. B., Andersson, B. & Lindberg, A. M. (2009). Molecular characterization of a novel Ljungan virus (Parechovirus; Picornaviridae) reveals a fourth genotype and indications of ancestral recombination. Journal of General Virology, 90, 843-853
Open this publication in new window or tab >>Molecular characterization of a novel Ljungan virus (Parechovirus; Picornaviridae) reveals a fourth genotype and indications of ancestral recombination
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2009 (English)In: Journal of General Virology, ISSN 0022-1317, E-ISSN 1465-2099, Vol. 90, p. 843-853Article in journal (Refereed) Published
Abstract [en]

Ljungan virus (LV) was discovered 20 years ago in Swedish bank voles (Myodes glareolus, previously referred to as Clethrionomys glareolus) during the search for an infectious agent causing lethal myocarditis in young athletes. To date, the genomes of four LV isolates, including the prototype 87-012 strain, have been characterized. Three of these LV strains were isolated from bank voles trapped in Sweden. Sequence analysis of an American virus (M1146), isolated from a montane vole (Microtus montanus) in western USA, indicates that this strain represents a genotype that is different from the Swedish strains. Here, we present genomic analyses of a fifth LV strain (64-7855) isolated from a southern red-backed vole (Myodes gapperi) trapped during arbovirus studies in New York state in the north-eastern USA in the 1960s. Sequence analysis of the 64-7855 genome showed an LV-like genome organization and sequence similarity to other LV strains. Genetic and phylogenetic analyses of the evolutionary relationship between the 64-7855 strain and other viruses within the family Picornaviridae, including previously published LV strains, demonstrated that the 64-7855 strain constitutes a new genotype within the LV species. Analyses also showed that different regions of the 64-7855 genome have different phylogenetic relationships with other LV strains, indicating that previous recombination events have been involved in the evolution of this virus.

National Category
Microbiology
Research subject
Biomedical Sciences, Virology; Natural Science, Microbiology; Natural Science, Biomedical Sciences
Identifiers
urn:nbn:se:lnu:diva-2140 (URN)10.1099/vir.0.007948-0 (DOI)
Note

The GenBank/EMBL/DDBJ accession number of the sequence reported in this paper isEU854568.

Available from: 2010-04-06 Created: 2010-04-06 Last updated: 2017-12-12Bibliographically approved
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