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1.
Int J Mol Sci ; 24(19)2023 Sep 26.
Article in English | MEDLINE | ID: mdl-37834041

ABSTRACT

Persistent high-risk human papillomavirus (HPV) infection is a pivotal factor in the progression of cervical cancer. In recent years, an increasing interest has emerged in comprehending the influence of HPV on head and neck squamous cell carcinoma (HNSCC). Notably, it is well established that HPV-associated HNSCC show cases with distinct molecular and clinical attributes compared to HPV-negative cases. The present study delves into the epigenetic landscape of HPV16, specifically its L1 gene and untranslated region (UTR), through pyrosequencing, while the HPV16 DNA physical status was evaluated using E2/E6 ratio analysis in HPV16-positive HNSCC FFPE biopsies. Our findings reveal substantial methylation across six sites within the HPV16 L1 gene and seven sites in the UTR. Specifically, methylation percentages of two L1 CpG sites (7136, 7145) exhibit significant associations with tumor histological grade (p < 0.01), while proving concurrent methylation across multiple sites. The HPV16 DNA physical status was not correlated with the methylation of viral genome or tumor characteristics. This is the first study that examines epigenetic modifications and the HPV16 DNA physical status in Greek HNSCC patients. Our findings suggest an orchestrated epigenetic modulation among specific sites, impacting viral gene expression and intricate virus-host interactions.


Subject(s)
Carcinoma, Squamous Cell , Head and Neck Neoplasms , Oncogene Proteins, Viral , Papillomavirus Infections , Female , Humans , Squamous Cell Carcinoma of Head and Neck/genetics , Squamous Cell Carcinoma of Head and Neck/complications , Human Papillomavirus Viruses , Carcinoma, Squamous Cell/pathology , DNA Methylation , Papillomavirus Infections/complications , Papillomavirus Infections/genetics , Head and Neck Neoplasms/genetics , Head and Neck Neoplasms/complications , DNA/metabolism , Oncogene Proteins, Viral/genetics , Oncogene Proteins, Viral/metabolism , DNA, Viral/genetics , DNA, Viral/metabolism
2.
Viruses ; 15(1)2022 12 31.
Article in English | MEDLINE | ID: mdl-36680181

ABSTRACT

The causal relationship between HPV and cervical cancer in association with the high prevalence of high risk HPV genotypes led to the design of HPV vaccines based on the major capsid L1 protein. In recent years, capsid protein L2 has also become a focal point in the field of vaccine research. The present review focuses on the variability of HPV16 L1 and L2 genes, emphasizing the distribution of specific amino acid changes in the epitopes of capsid proteins. Moreover, a substantial bioinformatics analysis was conducted to describe the worldwide distribution of amino acid substitutions throughout HPV16 L1, L2 proteins. Five amino acid changes (T176N, N181T; EF loop), (T266A; FG loop), (T353P, T389S; HI loop) are frequently observed in the L1 hypervariable surface loops, while two amino acid substitutions (D43E, S122P) are adjacent to L2 specific epitopes. These changes have a high prevalence in certain geographic regions. The present review suggests that the extensive analysis of the amino acid substitutions in the HPV16 L1 immunodominant loops may provide insights concerning the ability of the virus in evading host immune response in certain populations. The genetic variability of the HPV16 L1 and L2 epitopes should be extensively analyzed in a given population.


Subject(s)
Capsid Proteins , Oncogene Proteins, Viral , Humans , Amino Acids/genetics , Antibodies, Viral , Capsid Proteins/genetics , Epitopes , Human papillomavirus 16/genetics , Mutation , Oncogene Proteins, Viral/genetics , Papillomavirus Infections/virology
3.
Virology ; 526: 72-80, 2019 01 02.
Article in English | MEDLINE | ID: mdl-30366300

ABSTRACT

Recombination is a driving force for the emergence, evolution and virulence/epidemics of viruses, comprising the Enterovirus genus of the Picornaviridae family, important for human and animal health. By analyzing 2949 complete genomes/coding sequences, we provide a thorough and up-to-date overview of the genome-wide patterns and hotspots of intertypic recombination between the genogroups of this genus. Two prominent recombination hotspots are identified/verified, at the 5'UTR-capsid region junction, and at the beginning of the P2 region. In general, P2 was enriched in recombination events. Key phylogenetic groups implicated in recombination events are E71 and CVA6 in Enterovirus A species, E30 and E6 in Enterovirus B species, polioviruses 1 and 2 in Enterovirus C species. In addition, many events involve recombination partners that have not been sequenced yet, thus strongly suggesting a large environmental reservoir of genetic variation with a high potential for the emergence of new modified pathogens by recombination.


Subject(s)
Enterovirus/genetics , Evolution, Molecular , Genome, Viral/genetics , Recombination, Genetic , 5' Untranslated Regions/genetics , Capsid Proteins/genetics , Databases, Genetic , Enterovirus/classification , Genotype , Humans , Phylogeny , Rhinovirus/classification , Rhinovirus/genetics , Viral Nonstructural Proteins/genetics
4.
J Med Virol ; 90(5): 965-971, 2018 05.
Article in English | MEDLINE | ID: mdl-29168898

ABSTRACT

The tumor suppressor protein p16 plays a fundamental role in cell cycle regulation and exerts a protective effect against tumor growth. Two different polymorphisms at positions 540 and 580 at the 3'UTR of exon 3 of p16 gene are implicated in several types of cancer, while their role in cervical cancer development remains rather vague. In the present study, we investigated for the impact of p16 genotypes/haplotypes on patients' vulnerability to cervical disease and examined whether these factors can be used as progression markers in the Greek population. A total of 96 HPV16 positive samples and histologically confirmed as LSIL (42 samples), HSIL (44 samples), and cervical cancer cases (10 samples) along with 50 control cases were tested. The identification of p16 polymorphisms was performed by PCR-RFLP methodology. The present analysis revealed that women with p16 540 CG/GG genotype are at a 2.7-fold higher risk of developing HPV16-associated HSIL (OR = 2.7, 95%CI: 1.01-6.6, P = 0.028). The G allele can be regarded as a risk factor of developing HSIL in the Greek population (OR = 2.7, 95%CI: 1.2-5.9, P = 0.012). Moreover, p16 polymorphism C580T is not associated with the growth of cervical lesion in Greek patients, while 540G/580C haplotype can be regarded as a risk haplotype of developing HSIL (OR = 3.67, 95%CI: 1.56-8.6, P = 0.0019). Our results demonstrated that p16 C540G polymorphism influence patients' susceptibility to more severe dysplasia and consequently this polymorphism could potentially emerge as a valuable biomarker for HSIL development in the Greek population.


Subject(s)
Cyclin-Dependent Kinase Inhibitor p16/genetics , Genetic Predisposition to Disease , Human papillomavirus 16/isolation & purification , Squamous Intraepithelial Lesions of the Cervix/epidemiology , Squamous Intraepithelial Lesions of the Cervix/genetics , Uterine Cervical Neoplasms/epidemiology , Uterine Cervical Neoplasms/genetics , Adult , Female , Genotype , Greece/epidemiology , Humans , Middle Aged , Papillomavirus Infections/complications , Papillomavirus Infections/virology , Prospective Studies
5.
Arch Virol ; 163(2): 365-375, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29086105

ABSTRACT

Recombination has been recognized as a major mechanism of evolution in enteroviruses. The Echovirus 30 (E-30) strain Gior was sequenced and phylogenetically compared to all available E-30 sequences to detect recombination events between the 5΄UTR and VP1 genomic regions. The comparison of phylogenetic trees of the 5΄UTR and VP1 revealed incongruences concerning strains, lineages and sub-lineages. Comparative analysis of 62 E-30 sub-genomic sequences revealed six different recombination events that almost all occurred in the same region, having a start point in the 3΄end of the 5΄ UTR and end point in VP4. The only exception was the sub-lineage of Gior for which both borders of recombination were located in the 5΄UTR. These results describe for the first time recombination events in this region in circulating EV-B strains, revealing the exact points of these recombination events, highlighting the impact of such events on the evolution and epidemiology of enteroviruses.


Subject(s)
Echovirus Infections/virology , Enterovirus B, Human/genetics , Recombination, Genetic , Viral Proteins/genetics , 5' Untranslated Regions , Enterovirus B, Human/classification , Enterovirus B, Human/isolation & purification , Enterovirus B, Human/physiology , Evolution, Molecular , Genome, Viral , Humans , Phylogeny , RNA, Viral/genetics , Serogroup , Viral Proteins/metabolism
7.
J Med Microbiol ; 66(9): 1358-1365, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28857739

ABSTRACT

Purpose. Polymorphic variability in the tumour-suppressor protein p53 at codon 72 has a considerable impact on cervical cancer development. The present study clarified the association between p53 codon 72 genotypes and the risk of cervical disease in Greek patients. We also examined whether the presence of specific p53 genotypes in combination with HPV16 variants or E6 T350G sequence variation can modify an individual's susceptibility to cervical disease.Methodology. The analysis of p53 genotypes was performed through PCR-RFLP. Sequence and phylogenetic tree analyses of the HPV16 E6 gene were also performed in order to identify HPV16 variants and T350G sequence variation.Results/Key findings. The outcomes of the present analysis revealed that women who are homozygous for the arg genotype are at a 4.17-fold higher risk of developing HPV16-associated HSIL+ (OR=4.17, 95 % CI:1.48-4.9, P=0.0049). Moreover, p53 arg/arg patients infected by an HPV16 prototype strain were associated with an increased risk of more severe lesions, while a significant relationship between the p53 arg/arg genotype in patients with T350G sequence variation and the risk of high-grade squamous intraepithelial lesions (HSILs) was revealed.Conclusion. The oncogenic potential of the virus is increased by the presence of the p53 arg/arg genotype in the Greek population in such a way that the specific protein interaction E6 (L83V)-p53 (Arg-72) can modify an individual's susceptibility to cervical disease.

8.
BMC Bioinformatics ; 18(1): 13, 2017 Jan 05.
Article in English | MEDLINE | ID: mdl-28056784

ABSTRACT

BACKGROUND: Many computational tools that detect recombination in viruses are not adapted for the ongoing genomic revolution. A computational tool is needed, that will rapidly scan hundreds/thousands of genomes or sequence fragments and detect candidate recombination events that may later be further analyzed with more sensitive and specialized methods. RESULTS: T-RECs, a Windows based graphical tool, employs pairwise alignment of sliding windows and can perform (i) genotyping, (ii) clustering of new genomes, (iii) detect recent recombination events among different evolutionary lineages, (iv) manual inspection of detected recombination events by similarity plots and (v) annotation of genomic regions. CONCLUSIONS: T-RECs is very effective, as demonstrated by an analysis of 555 Norovirus complete genomes and 2500 sequence fragments, where a recombination hotspot was identified at the ORF1-ORF2 junction.


Subject(s)
Genome, Viral , Norovirus/genetics , Recombination, Genetic , Cluster Analysis , DNA Fragmentation , DNA, Viral/genetics , Databases, Genetic , Evolution, Molecular , Genotyping Techniques , Sequence Alignment , Sequence Analysis, DNA , Software
9.
Viral Immunol ; 29(7): 444-50, 2016 09.
Article in English | MEDLINE | ID: mdl-27410516

ABSTRACT

Mutations and recombination events have been identified in enteroviruses. Point mutations accumulate with a frequency of 6.3 × 10(-4) per base pair per replication cycle affecting the fitness, the circulation, and the infectivity of enteroviral strains. In the present report, the serological status of the Central and Western Greek population (Larissa and Ioannina, respectively) in the 1-10-year, 11-20-year, 21-30-year, and 31-40-year age groups against six non-polio enterovirus strains, their respective echovirus prototypes, and Sabin 1, 2, and 3 vaccine strains was evaluated, through serum-neutralization assay. In the Western Greek population, antibody levels were detected only for clinical isolates of E30 serotype in all age groups, and for environmental isolate LR61G3 (E6 serotype) only in the 31-40 age group, whereas an immunity level was observed in the Central Greek population, against all strains, except for EIS6B (E3 serotype). Amino acid substitutions were encountered across the structural region of the capsid, between the prototypes and the respective isolates. These substitutions may alter the antigenicity of each strain and may explain the variations observed in the neutralization titers of the different strains. As a consequence, these substitutions severely affect antibody binding and increase the ability of the virus to escape the immune response. It is tempting to assume that changes in the antigenic properties observed in circulating echoviruses represent a selection of viral variants that are less prone to be neutralized by human antibodies. These facts argue for the need of immunological studies to the population to avoid epidemics due to the circulation of highly evolved derivatives.


Subject(s)
Antibodies, Neutralizing/blood , Antibodies, Viral/blood , Enterovirus Infections/epidemiology , Enterovirus/immunology , Neutralization Tests/methods , Adolescent , Adult , Amino Acid Substitution , Capsid Proteins/genetics , Capsid Proteins/immunology , Child , Child, Preschool , Enterovirus/genetics , Enterovirus Infections/immunology , Greece/epidemiology , Humans , Infant , Seroepidemiologic Studies , Young Adult
10.
Infect Genet Evol ; 32: 305-12, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25791929

ABSTRACT

Echovirus 3 (E3) serotype has been related with several neurologic diseases, although it constitutes one of the rarely isolated serotypes, with no report of epidemics in Europe. The aim of the present study was to provide insights into the molecular epidemiology and evolution of this enterovirus serotype, while an E3 strain was isolated from sewage in Greece, four years after the initial isolation of the only reported E3 strain in the same geographical region. Phylogenetic analysis of the complete VP1 genomic region of that E3 strain and of those available in GenBank suggested three main genogroups that were further subdivided into seven subgenogroups. Further evolutionary analysis suggested that VP1 genomic region of E3 was dominated by purifying selection, as the vast majority of genetic diversity presumably occurred through synonymous nucleotide substitutions and the substitution rate for complete and partial VP1 sequences was calculated to be 8.13×10(-3) and 7.72×10(-3) substitutions/site/year respectively. The partial VP1 sequence analysis revealed the composite epidemiology of this serotype, as the strains of the three genogroups presented different epidemiological characteristics.


Subject(s)
Echovirus Infections/epidemiology , Enterovirus/genetics , Evolution, Molecular , Molecular Epidemiology , Serogroup , Databases, Genetic , Enterovirus/classification , Enterovirus/isolation & purification , Genetic Variation , Genotype , Greece/epidemiology , Multigene Family , Phylogeny , RNA, Viral/genetics
11.
Virus Genes ; 50(2): 177-88, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25537948

ABSTRACT

Human enteroviruses (EV) belong to the Picornaviridae family and are among the most common viruses infecting humans. They consist of up to 100 immunologically and genetically distinct types: polioviruses, coxsackieviruses A and B, echoviruses, and the more recently characterized 43 EV types. Frequent recombinations and mutations in enteroviruses have been recognized as the main mechanisms for the observed high rate of evolution, thus enabling them to rapidly respond and adapt to new environmental challenges. The first signs of genetic exchanges between enteroviruses came from polioviruses many years ago, and since then recombination has been recognized, along with mutations, as the main cause for reversion of vaccine strains to neurovirulence. More recently, non-polio enteroviruses became the focus of many studies, where recombination was recognized as a frequent event and was correlated with the appearance of new enterovirus lineages and types. The accumulation of multiple inter- and intra-typic recombination events could also explain the series of successive emergences and disappearances of specific enterovirus types that could in turn explain the epidemic profile of circulation of several types. This review focuses on recombination among human non-polio enteroviruses from all four species (EV-A, EV-B, EV-C, and EV-D) and discusses the recombination effects on enterovirus epidemiology and evolution.


Subject(s)
Enterovirus C, Human/genetics , Enterovirus Infections/virology , Evolution, Molecular , Recombination, Genetic , Animals , Enterovirus C, Human/classification , Enterovirus C, Human/isolation & purification , Enterovirus Infections/epidemiology , Humans
12.
J Med Microbiol ; 63(Pt 12): 1716-1723, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25212758

ABSTRACT

Human papillomavirus (HPV) 16 genome integration into the host chromosome is a crucial event during the life cycle of the virus and a major step towards carcinogenesis. The integration of HPV16 DNA promotes a constitutive high expression level of E6 and E7 oncoproteins, resulting in the extensive proliferation of the infected epithelial cells. In the present report the physical status of the HPV16 genome was studied, through determination of E1/E6 and E2/E6 DNA copy number ratios in 61 cervical samples of low- and high-grade malignancy and 8 cervical cancer samples, all of them associated with HPV16 infection. The selection of E1, E2 and E6 amplification target regions was performed according to the most prevalent deleted/disrupted sites of E1 and E2 genes. For this target selection we also considered the most conserved regions of E1, E2 and E6 genes among the same HPV16 isolates that were recently reported by our group. The analysis of HPV16 DNA form revealed a significant association among the mixed DNA forms in low-grade and high-grade malignancies, (χ(2), P<0.01). The comparative analysis of E1/E6 and E2/E6 in the same cervical samples provides an accurate picture of HPV16 DNA form and may reveal whether different HPV16 DNA integrants coexist in the same cervical sample or not. This study proposes that E1/E6 and E2/E6 ratios determine with accuracy the HPV16 DNA integration pattern and may predict multiple integration events in the examined sample, thus providing significant information about the progression of cervical dysplasia.


Subject(s)
Cervix Uteri/virology , DNA, Viral/analysis , DNA-Binding Proteins/genetics , Human papillomavirus 16/physiology , Oncogene Proteins, Viral/genetics , Repressor Proteins/genetics , Virus Integration , DNA, Viral/genetics , Female , Gene Dosage , Human papillomavirus 16/genetics , Humans , Papillomavirus Infections/virology
13.
Arch Virol ; 158(1): 103-11, 2013 Jan.
Article in English | MEDLINE | ID: mdl-22983155

ABSTRACT

Enteroviruses, the main cause of aseptic meningitis, consist of 100 serotypes, and many of them have been associated with large outbreaks. In the present study, a comparison of RFLP analysis of the 5' untranslated region (5'UTR) and sequencing of both the 5'UTR and VP1 regions was conducted for epidemiological linkage of 27 clinical enterovirus strains. The clinical enterovirus strains were clustered into five restriction profile groups. Even though the restriction profile clusters of clinical isolates were not related to those of the respective prototype strains, epidemiological relationships between the members of each cluster were observed. The restriction profile clusters in the 5'UTR corresponded to the phylogenetic clusters in the VP1 genomic region. The incongruence between the topology of Gior strain in 5'UTR and VP1 phylogenetic trees indicates a recombination event. The proposed RFLP assay in combination with VP1 sequencing can offer crucial epidemiological information about the circulating enteroviruses.


Subject(s)
5' Untranslated Regions , Enterovirus Infections/virology , Enterovirus/genetics , Polymorphism, Restriction Fragment Length , Viral Envelope Proteins/genetics , Enterovirus/classification , Enterovirus/isolation & purification , Enterovirus Infections/epidemiology , Epidemics , Greece/epidemiology , Humans , Molecular Sequence Data , Phylogeny
14.
Expert Rev Vaccines ; 11(5): 609-28, 2012 May.
Article in English | MEDLINE | ID: mdl-22827246

ABSTRACT

The Global Polio Eradication Initiative was launched in 1988 with the aim to eliminate paralytic poliomyelitis. Two effective vaccines are available: inactivated polio vaccine (IPV) and oral polio vaccine (OPV). Since 1964, OPV has been used instead of IPV in most countries due to several economic and biological advantages. However, in rare cases, the live-attenuated Sabin strains of OPV revert to neurovirulence and cause vaccine-associated paralytic poliomyelitis in vaccinees or lead to emergence of vaccine-derived poliovirus strains. Attenuating mutations and recombination events have been associated with the reversion of vaccine strains to neurovirulence. The substitution of OPV with an improved new-generation IPV and the availability of new specific drugs against polioviruses are considered as future strategies for outbreak control and the eradication of paralytic poliomyelitis worldwide.


Subject(s)
Poliomyelitis/prevention & control , Poliovirus Vaccine, Inactivated/adverse effects , Poliovirus Vaccine, Oral/adverse effects , Adolescent , Adult , Child , Child, Preschool , Disease Outbreaks/prevention & control , Humans , Infant , Mutation , Poliomyelitis/virology , Poliovirus/genetics , Poliovirus/immunology , Poliovirus/pathogenicity , Poliovirus Vaccine, Inactivated/therapeutic use , Poliovirus Vaccine, Oral/therapeutic use , Risk , Young Adult
15.
Virus Genes ; 44(2): 207-16, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22090198

ABSTRACT

Echovirus 6 (E6) is one of the main enteroviral serotypes that was isolated from cases of aseptic meningitis and encephalitis during the last years in Greece. Two E6 (LR51A5 and LR61G3) were isolated from the sewage treatment plant unit in Larissa, Greece, in May 2006, 1 year before their characterization from aseptic meningitis cases. The two isolates were initially found to be intra-serotypic recombinants in the genomic region VP1, a finding that initiated a full genome sequence analysis. In the present study, nucleotide, amino acid, and phylogenetic analyses for all genomic regions were conducted. For the detection of recombination events, Simplot and bootscan analyses were carried out. The continuous phylogenetic relationship in 2C-3D genomic region of strains LR51A5 and LR61G3 with E30 isolated in France in 2002-2005 indicated that the two strains were recombinants. SimPlot and Bootscan analyses confirmed that LR51A5 and LR61G3 carry an inter-serotypic recombination in the 2C genomic region. The present study provide evidence that recombination events occurred in the regions VP1 (intraserotypic) and non-capsid (interserotypic) during the evolution of LR51A5 and LR61G3, supporting the statement that the genomes of circulating enteroviruses are a mosaic of genomic regions of viral strains of the same or different serotypes. In conclusion, full genome sequence analysis of circulating enteroviral strains is a prerequisite to understand the complexity of enterovirus evolution.


Subject(s)
Echovirus 6, Human/genetics , Echovirus 6, Human/isolation & purification , Genome, Viral , RNA, Viral/genetics , Sewage/virology , Cluster Analysis , Echovirus 6, Human/classification , Greece , Humans , Molecular Sequence Data , Phylogeny , Sequence Analysis, DNA
16.
Vaccine ; 29(1): 26-33, 2010 Dec 10.
Article in English | MEDLINE | ID: mdl-20974307

ABSTRACT

In this study, the immunity level of the southern Greek population in the 1-10-year, 11-20-year, 21-30-year and 31-40-year age groups with regard to Sabin vaccine strains and a collection of 11 recombinant and three non-recombinant poliovirus vaccine strains was determined. The results showed the lowest neutralization titre in the 21-30-year-age group against poliovirus type 3. Moreover, the capsid coding region of OPV (oral poliovirus vaccine) derivatives was sequenced in order to identify mutations that might lead to antigenic changes. In Sabin-1 derivatives a tendency of accumulation of mutations was observed in or near antigenic sites while in Sabin-2 and Sabin-3 derivatives in sites known to be involved in restoring neurovirulence or eliminating their temperature-sensitive phenotype. It was concluded that the combination of mutations in the capsid coding region and not the number of specific mutations in antigenic sites determines the antigenic properties of OPV derivatives and their reactivity with human sera.


Subject(s)
Antibodies, Neutralizing/blood , Antibodies, Viral/blood , Antigens, Viral/immunology , Poliomyelitis/prevention & control , Poliovirus Vaccine, Oral/immunology , Poliovirus/immunology , Adolescent , Adult , Antigens, Viral/genetics , Child , Child, Preschool , Greece , Humans , Infant , Molecular Sequence Data , Mutation, Missense , Poliomyelitis/immunology , Poliovirus/genetics , Poliovirus Vaccine, Oral/genetics , RNA, Viral/genetics , Sequence Analysis, DNA , Viral Proteins/genetics , Viral Proteins/immunology , Young Adult
17.
Virus Genes ; 41(2): 181-91, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20640496

ABSTRACT

In order to test the hypothesis that RNA structural elements promote the distribution of certain types of recombination junctions in each one of the 2C and 3D poliovirus genomic regions (Sabin 3/Sabin 2 or Sabin 1 in 2C and Sabin 2/Sabin 1 or Sabin 3 in 3D), we searched in 2C and 3D regions of reference Sabin strains for high probability RNA structural elements that could promote recombination. Recombination junctions that were identified in clinical strains of this study, as well as in clinical strains of previous studies, were superimposed on RNA secondary structure models of 2C and 3D genomic regions. Furthermore, we created an in vitro model, based on double infection of cell-culture with two poliovirus strains, for the production and identification of recombinant Sabin strains in 2C and 3D regions. Our intention was to compare the results that refer to the correlation of recombination junctions and RNA secondary structures in 2C and 3D regions of clinical strains, with the respective results of the in vitro model. Most of the recombination junctions of the clinical strains were correlated with RNA secondary structure elements, which were identical between recombining Sabin strains, and also presented high predictive value. In consensus were, the respective results originated from the in vitro model. We propose that the distribution of specific types of recombination junctions in certain regions of Sabin strains is not fortuitous and is correlated with RNA secondary structure elements identical to both recombination partners. Furthermore, results of this study highlight an important role for the stem region of the RNA structure elements in promoting recombination.


Subject(s)
Nucleic Acid Conformation , Poliovirus/genetics , RNA, Viral/genetics , Recombination, Genetic , DNA Fingerprinting , Humans , Models, Molecular , Molecular Sequence Data , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNA
18.
J Clin Microbiol ; 48(5): 1513-9, 2010 May.
Article in English | MEDLINE | ID: mdl-20129960

ABSTRACT

An echovirus 3 (Echo3) strain (strain LR31G7) was isolated from a sewage treatment plant in Greece in 2005. Full-genome molecular, phylogenetic, and SimPlot analyses were conducted in order to reveal the evolutionary pathways of the isolate. Nucleotide and phylogenetic analyses of part of the VP1 genomic region revealed that the isolated strain correlates with Echo3 strains isolated during the same year in France and Japan, implying that the same virus circulated in Europe and Asia. LR31G7 was found to be a recombinant that shares the 3' part of its genome with an Echo25 strain isolated from asymptomatic infants in Norway in 2003. Nucleotide and SimPlot analyses of the VP1-2A junction, where the recombination was located, revealed the exact recombination breakpoint (nucleotides 3357 to 3364). Moreover, there is evidence that recombination events had occurred in 3B-3D region in the evolutionary history of the isolate. Our study indicates that recombination events play major roles in enterovirus evolution and that the circulation of multirecombinant strains with unknown properties could be potentially dangerous for public health.


Subject(s)
Enterovirus B, Human/genetics , Genome, Viral , Recombination, Genetic , Sequence Analysis, DNA , Sewage/virology , Cluster Analysis , Enterovirus B, Human/isolation & purification , Evolution, Molecular , Greece , Humans , Molecular Sequence Data , Phylogeny , RNA, Viral/genetics , Viral Structural Proteins/genetics
19.
Virus Genes ; 40(2): 183-92, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20094765

ABSTRACT

Two enteroviruses from river water and four from sewage treatment plant were isolated in Larissa, Greece, that all shared the same sequence. A full genome analysis was conducted in an attempt to reveal the evolutionary pathways of one of the isolated strains (LR11F7). VP1 nucleotide and phylogenetic analysis revealed that the isolated strain had 78% homology with the echovirus 7 prototype strain Wallace. Full genome analysis revealed that LR11F7 P1 region is related to echoviruses 7 and that P2 and P3 regions are originating from contemporary enteroviruses isolated in South Asia. Two recombination events were shown to be involved into the evolutionary history of LR11F7, the one event concerning 3A, 3B, and 2C, and the other concerning 3D genomic region, both with new types of HEV-B. The contribution of recombination to enterovirus evolution is substantial, giving rise to new genetic lineages with unknown properties.


Subject(s)
Enterovirus B, Human/genetics , Enterovirus B, Human/isolation & purification , Fresh Water/virology , Genome, Viral , RNA, Viral/genetics , Capsid Proteins/genetics , Cluster Analysis , Enterovirus B, Human/classification , Genotype , Greece , Humans , Molecular Sequence Data , Phylogeny , Recombination, Genetic , Rivers , Sequence Analysis, DNA , Sequence Homology, Amino Acid
20.
Virus Genes ; 40(2): 200-11, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20091423

ABSTRACT

Attenuated strains of Sabin poliovirus vaccine replicate in the human gut and in rare cases may cause vaccine-associated paralytic poliomyelitis (VAPP). Mutations at specific sites of the genome and recombination between Sabin strains may result in the loss of the attenuated phenotype of OPV (Oral Poliovirus Vaccine) strains and the acquisition of traits characteristic of wild polioviruses, such as increased neurovirulence and loss of temperature sensitivity. In this study, we determined the phenotypic traits such as temperature sensitivity and growth kinetics of eight OPV isolates (six bi-recombinant and two non-recombinant). The growth phenotype of each isolate as well as of Sabin vaccine strains in Hep2 cell line at two different temperatures (37 and 40 degrees C) was evaluated using two different assays, RCT test (Reproductive Capacity at different Temperatures) and one-step growth curve analysis. Moreover, the nucleotide and amino acid positions in the genomes of the isolates that have been identified as being involved in the attenuated and thermo sensitive phenotype of Sabin vaccine strains were investigated. Mutations that result in loss of the attenuated and thermo sensitive phenotype of Sabin vaccine strains were identified in the genomes of all isolates. Both mutations and recombination events correlated well with the reverted phenotypic traits of OPV-derivatives. In the post-eradication era of wild polioviruses, the identification and the characterization (genomic and phenotypic) of vaccine-derived polioviruses become increasingly important in order to prevent cases or even outbreaks of paralytic poliomyelitis caused by neurovirulent strains.


Subject(s)
Poliovirus Vaccines , Poliovirus/growth & development , Poliovirus/genetics , Cell Line , DNA Mutational Analysis , Hepatocytes/virology , Humans , Poliovirus/pathogenicity , RNA, Viral/genetics , Recombination, Genetic , Suppression, Genetic , Temperature , Vaccines, Attenuated , Virulence
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