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1.
Sci Rep ; 11(1): 6369, 2021 03 18.
Article in English | MEDLINE | ID: mdl-33737658

ABSTRACT

O'nyong-nyong virus is an alphavirus closely related to chikungunya virus, causing arthralgia, rash and fever. Alphaviruses mainly target synovial fibroblasts and persists in the joints of patients, possibly leading to chronic arthritis. To date, no specific antiviral treatment is available for ONNV infection and induced-inflammation. Primary human synovial fibroblasts cells were used to assess infection by ONNV and the resulting cytokine responses. Phenolics (gallic acid, caffeic acid and chlorogenic acid, curcumin and quercetin) and a curcuminoids-rich extract from turmeric were tested for their antiviral and anti-inflammatory capacities. We showed that infection occurred in HSF cells and increased gene expression and protein secretion of two major proinflammatory CCL-2 and IL-1ß markers. In ONNV-infected HSF cells (MOI 1), we found that non-cytotoxic concentrations of phenolics (10 µM) reduced the level of viral RNA (E1, E2, nsP1, nsP2) and downregulated CCL-2 and IL-1ß expression and secretion. These results highlighted the high value of the flavonol quercetin to reduce viral RNA levels and inflammatory status induced by ONNV in HSF cells.


Subject(s)
Alphavirus Infections/drug therapy , Chemokine CCL2/genetics , Immunity, Innate/genetics , Interleukin-1beta/genetics , Alphavirus Infections/genetics , Alphavirus Infections/pathology , Alphavirus Infections/virology , Caffeic Acids/pharmacology , Chlorogenic Acid/pharmacology , Curcumin/pharmacology , Cytokines/genetics , Fibroblasts/virology , Gallic Acid/pharmacology , Humans , Immunity, Innate/drug effects , O'nyong-nyong Virus/genetics , O'nyong-nyong Virus/pathogenicity , Quercetin/pharmacology , Synovial Fluid/drug effects , Synovial Fluid/virology
2.
Avian Pathol ; 48(4): 334-342, 2019 Aug.
Article in English | MEDLINE | ID: mdl-30915860

ABSTRACT

Avian orthoreovirus (ARV) infections of broiler flocks cause arthritis/tenosynovitis syndrome and significant economic losses. ARV variants were detected in the USA and Canada. Viral arthritis/tenosynovitis syndrome has occurred frequently in China in recent years. In this study, a variant ARV strain associated with viral arthritis/tenosynovitis syndrome was isolated from broilers and designated as LY383. Genomic sequence and phylogenetic analysis of the σC nucleic acid and amino acid sequences revealed that the isolate was closely related to ARV field strains Reo/PA/Layer/01224B/14, Reo/PA/Broiler/1551/13, GA/14602/2014, GA/13569/2013 and GA/13542/2013, in cluster V, but distinct from most Chinese field strains or commercial vaccine strains. Experimental challenge showed that the isolate could cause arthritis/tenosynovitis syndrome in broilers, which possessed a high level of maternal antibodies induced by commercial ARV vaccines (S1133, 1733 and T98). Furthermore, viral nucleic acid could be detected in cloacal swabs of all challenged birds throughout the entire test from 5 dpi onward. These results suggest that a novel ARV genotype emerges and might become prevalent in broiler flocks in China. RESEARCH HIGHLIGHTS A variant avian orthoreovirus was isolated from a vaccinated broiler flock in North China. The ARV field strain was distinct from previous China-origin ARV isolates and vaccine strains. The current commercial ARV vaccine could not provide effective protection of broilers against the field isolate infection. These findings indicated that variant ARV field strains might become frequent in broiler flocks in China and effective measures should be conducted to prevent and control the disease.


Subject(s)
Chickens , Genome/genetics , Orthoreovirus, Avian/genetics , Orthoreovirus, Avian/pathogenicity , Poultry Diseases/virology , Reoviridae Infections/veterinary , Amino Acid Sequence , Animals , Arthritis/veterinary , Capsid Proteins/chemistry , Capsid Proteins/genetics , China , High-Throughput Nucleotide Sequencing/veterinary , Orthoreovirus, Avian/classification , Phylogeny , Poultry Diseases/prevention & control , RNA, Viral/chemistry , RNA-Binding Proteins/chemistry , RNA-Binding Proteins/genetics , Random Allocation , Reoviridae Infections/prevention & control , Reoviridae Infections/virology , Synovial Fluid/virology , Tendons/virology , Tenosynovitis/veterinary , Vaccination/veterinary
3.
PLoS One ; 13(12): e0208957, 2018.
Article in English | MEDLINE | ID: mdl-30533036

ABSTRACT

OBJECTIVE: The objective of the present study was to investigate Epstein-Barr virus (EBV) infection as an environmental factor for the development of rheumatoid arthritis (RA). METHODS: Synovial tissues were collected during surgery from 128 RA and 98 osteoarthritis (OA) patients. DNA was extracted from synovial tissues. The EBV gene was assessed by nested PCR for the amplification of EBV nuclear antigen-1 (EBNA-1). The nucleotide sequence of the PCR product was elucidated. HLA-DRB1 genotyping was also performed. RESULTS: EBV DNA was more frequently detected in the synovial tissues of RA patients (32.8%) than OA patients (15.3%) (p<0.01). The frequency of EBNA-1 variants did not significantly differ between RA and OA (RA: 17%, OA: 13%). The population with the HLA-DRB1 shared epitope (SE) was significantly higher in RA patients (70.3%) than in OA patients (44.9%) (p<0.001). In RA patients, the presence of EBV DNA was similar among SE-positive and -negative patients (SE-positive: 34.4%, -negative: 28.9%). The population with the EBNA-1 variant did not significantly differ between SE-positive and -negative patients (SE-positive: 12.9%, -negative: 27.3%). DISCUSSION: The present results indicate that EBV infection contributes to the onset of RA and chronic inflammation in synovial tissues. The frequency of EBNA-1 gene variants was low and not significantly different between RA and OA, suggesting that EBNA-1 gene variants are not a risk factor for RA. HLA-DRB1 with SE is a genetic risk factor for the development of RA. However, neither the presence of EBV nor EBNA-1 gene variants differed between SE-positive and -negative RA patients. Therefore, these two risk factors, SE and EBV, may be independent. CONCLUSION: EBV infection may be an environmental risk factor for the development of RA, while nucleotide variants of EBNA-1 do not appear to contribute to its development.


Subject(s)
Arthritis, Rheumatoid/virology , Epstein-Barr Virus Infections/genetics , Epstein-Barr Virus Nuclear Antigens/immunology , Herpesvirus 4, Human/genetics , Arthritis, Rheumatoid/genetics , Arthritis, Rheumatoid/pathology , Epitopes/genetics , Epitopes/immunology , Epstein-Barr Virus Infections/physiopathology , Epstein-Barr Virus Infections/virology , Epstein-Barr Virus Nuclear Antigens/genetics , Epstein-Barr Virus Nuclear Antigens/isolation & purification , Female , HLA-DRB1 Chains/genetics , HLA-DRB1 Chains/immunology , Herpesvirus 4, Human/isolation & purification , Herpesvirus 4, Human/pathogenicity , Humans , Male , Osteoarthritis/genetics , Osteoarthritis/physiopathology , Osteoarthritis/virology , Risk Factors , Synovial Fluid/virology
4.
Arthritis Rheumatol ; 70(4): 585-593, 2018 04.
Article in English | MEDLINE | ID: mdl-29266856

ABSTRACT

OBJECTIVE: To determine if chikungunya virus persists in synovial fluid after infection, potentially acting as a causative mechanism of persistent arthritis. METHODS: We conducted a cross-sectional study of 38 Colombian participants with clinical chikungunya virus infection during the 2014-2015 epidemic who reported chronic arthritis and 10 location-matched controls without chikungunya virus or arthritis. Prior chikungunya virus infection status was serologically confirmed, and the presence of synovial fluid chikungunya virus, viral RNA, and viral proteins was determined by viral culture, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and mass spectrometry, respectively. Biomarkers were assessed by multiplex analysis. RESULTS: Patients with serologically confirmed chikungunya arthritis (33 of 38 [87%]) were predominantly female (82%) and African Colombian (55%) or white Colombian (33%), with moderate disease activity (mean ± SD Disease Activity Score in 28 joints 4.52 ± 0.77) a median of 22 months after infection (interquartile range 21-23 months). Initial symptoms of chikungunya virus infection included joint pain (97%), swelling (97%), stiffness (91%), and fever (91%). The most commonly affected joints were the knees (87%), elbows (76%), wrists (75%), ankles (56%), fingers (56%), and toes (56%). Synovial fluid samples from all patients with chikungunya arthritis were negative for chikungunya virus on qRT-PCR, showed no viral proteins on mass spectrometry, and cultures were negative. Case and control plasma cytokine and chemokine concentrations did not differ significantly. CONCLUSION: This is one of the largest observational studies involving analysis of the synovial fluid of chikungunya arthritis patients. Synovial fluid analysis revealed no detectable chikungunya virus. This finding suggests that chikungunya virus may cause arthritis through induction of potential host autoimmunity, suggesting a role for immunomodulating agents in the treatment of chikungunya arthritis, or that low-level viral persistence exists in synovial tissue only and is undetectable in synovial fluid.


Subject(s)
Arthritis, Infectious/metabolism , Chikungunya Fever/metabolism , Chikungunya virus/metabolism , Synovial Fluid/virology , Arthritis, Infectious/virology , Chikungunya Fever/virology , Cross-Sectional Studies , Female , Humans , Male , Time Factors
6.
Clin Rheumatol ; 34(10): 1681-6, 2015 Oct.
Article in English | MEDLINE | ID: mdl-25980837

ABSTRACT

Viral agents have been suspected as participants of immune-mediated disorders. In the case of rheumatic diseases, the synovial joint cavity represents a secluded area of inflammation which could harbor etiological agents. We analyzed by polymerase chain reaction the possible presence of DNA from various herpes viruses in blood and synovial fluid from patients with either rheumatoid arthritis (n = 18), axial spondyloarthritis (n = 11), or osteoarthritis (n = 8). Relevant findings were as follows: DNA from varicella zoster virus was found in synovial fluid but not in blood mononuclear cells from 33 % of patients with rheumatoid arthritis and in 45 % of patients with axial spondyloarthritis but not in patients with osteoarthritis. Also, DNA from herpes simplex viruses 1 and 2 was found both in the blood and in the synovial fluid from 33 % of patients with rheumatoid arthritis. Our results indicate the occasional presence of DNA from herpes viruses in patients with rheumatoid arthritis or with axial spondyloarthritis. However, these findings might represent a parallel epiphenomenon of viral activation associated either with immunosuppressive therapy or with primary immune disturbances, rather than the etiological participation of herpes viruses in these disorders.


Subject(s)
Arthritis, Rheumatoid/blood , Arthritis, Rheumatoid/virology , Herpesviridae , Spondylarthritis/blood , Spondylarthritis/virology , Synovial Fluid/virology , Adult , Aged , Antibodies, Viral/analysis , Cross-Sectional Studies , DNA, Viral/analysis , Female , Herpesvirus 1, Human , Herpesvirus 2, Human , Herpesvirus 3, Human , Herpesvirus 4, Human , Herpesvirus 6, Human , Humans , Immunoglobulin G/analysis , Immunoglobulin M/analysis , Leukocytes, Mononuclear/virology , Male , Middle Aged , Osteoarthritis/virology , Real-Time Polymerase Chain Reaction , Young Adult
7.
Biomed Res Int ; 2014: 698609, 2014.
Article in English | MEDLINE | ID: mdl-25136615

ABSTRACT

The etiology of viruses in osteoarthritis remains controversial because the prevalence of viral nucleic acid sequences in peripheral blood or synovial fluid from osteoarthritis patients and that in healthy control subjects are similar. Until now the presence of virus has not been analyzed in cartilage. We screened cartilage and chondrocytes from advanced and non-/early osteoarthritis patients for parvovirus B19, herpes simplex virus-1, Epstein Barr virus, cytomegalovirus, human herpes virus-6, hepatitis C virus, and human endogenous retroviruses transcripts. Endogenous retroviruses transcripts, but none of the other viruses, were detected in 15 out the 17 patients. Sequencing identified the virus as HERV-WE1 and E2. HERV-W activity was confirmed by high expression levels of syncytin, dsRNA, virus budding, and the presence of virus-like particles in all advanced osteoarthritis cartilages examined. Low levels of HERV-WE1, but not E2 envelope RNA, were observed in 3 out of 8 non-/early osteoarthritis patients, while only 3 out of 7 chondrocytes cultures displayed low levels of syncytin, and just one was positive for virus-like particles. This study demonstrates for the first time activation of HERV-W in cartilage of osteoarthritis patients; however, a causative role for HERV-W in development or deterioration of the disease remains to be proven.


Subject(s)
Endogenous Retroviruses/genetics , Endogenous Retroviruses/isolation & purification , Gene Products, env/genetics , Osteoarthritis/virology , Pregnancy Proteins/genetics , Adult , Aged , Aged, 80 and over , Cartilage/pathology , Cartilage/virology , Chondrocytes/pathology , Chondrocytes/virology , Endogenous Retroviruses/pathogenicity , Female , Gene Products, env/isolation & purification , Humans , Male , Middle Aged , Osteoarthritis/blood , Osteoarthritis/pathology , Pregnancy Proteins/isolation & purification , RNA, Double-Stranded/genetics , RNA, Double-Stranded/isolation & purification , Synovial Fluid/virology
8.
J Med Virol ; 84(8): 1267-73, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22711355

ABSTRACT

Human bocaviruses 1-4 (HBoV1-4) and parvovirus 4 (PARV4) are recently discovered human parvoviruses. HBoV1 is associated with respiratory infections of young children, while HBoV2-4 are enteric viruses. The clinical manifestations of PARV4 remain unknown. The objective of this study was to determine whether the DNAs of HBoV1-4 and PARV4 persist in human tissues long after primary infection. Biopsies of tonsillar tissue, skin, and synovia were examined for HBoV1-4 DNA and PARV4 DNA by PCR. Serum samples from the tissue donors were assayed for HBoV1 and PARV4 IgG and IgM antibodies. To obtain species-specific seroprevalences for HBoV1 and for HBoV2/3 combined, the sera were analyzed after virus-like particle (VLP) competition. While HBoV1 DNA was detected exclusively in the tonsillar tissues of 16/438 individuals (3.7%), all of them ≤8 years of age. HBoV2-4 and PARV4 DNAs were absent from all tissue types. HBoV1 IgG seroprevalence was 94.9%. No subject had HBoV1 or PARV4 IgM, nor did they have PARV4 IgG. The results indicate that HBoV1 DNA occurred in a small proportion of tonsils of young children after recent primary HBoV1 infection, but did not persist long in the other tissue types studied, unlike parvovirus B19 DNA. The results obtained by the PARV4 assays are in line with previous results on PARV4 epidemiology.


Subject(s)
Human bocavirus/isolation & purification , Palatine Tonsil/virology , Parvoviridae Infections/epidemiology , Parvovirus/isolation & purification , Skin/virology , Synovial Fluid/virology , Adolescent , Adult , Aged , Biopsy , Child , Child, Preschool , DNA, Viral/analysis , Human bocavirus/genetics , Human bocavirus/immunology , Humans , Immunoglobulin G/blood , Immunoglobulin M/blood , Infant , Middle Aged , Parvoviridae Infections/immunology , Parvoviridae Infections/virology , Parvovirus/genetics , Parvovirus/immunology , Polymerase Chain Reaction , Species Specificity , Tonsillitis/epidemiology , Tonsillitis/immunology , Tonsillitis/virology , Young Adult
9.
Autoimmun Rev ; 11(11): 804-5, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22349617

ABSTRACT

In 1995, the SLAM-associated protein (SAP) gene was cloned in our laboratory, and patents were subsequently filed in the USA, EU and Canada. In 1998, the SAP gene was shown to be defective in patients with X-linked lymphoproliferative disease. Since this discovery, the gene has been demonstrated to play a significant role in viral protection, cytokine production and life-long immune memory (vaccination). This article reviews the crucial role of SAP in the development of autoimmunity.


Subject(s)
Autoimmunity/immunology , Intracellular Signaling Peptides and Proteins/physiology , Animals , Arthritis, Rheumatoid/immunology , Arthritis, Rheumatoid/virology , Autoantibodies/immunology , Autoantibodies/metabolism , Autoimmunity/genetics , Herpesvirus 4, Human/immunology , Humans , Intracellular Signaling Peptides and Proteins/genetics , Intracellular Signaling Peptides and Proteins/metabolism , Signaling Lymphocytic Activation Molecule Associated Protein , Synovial Fluid/virology
10.
Rheumatol Int ; 32(7): 1885-91, 2012 Jul.
Article in English | MEDLINE | ID: mdl-21445545

ABSTRACT

The SV40 T antigen has been used to generate immortalized cells from rheumatoid arthritis (RA) synovial fibroblasts (RASFs) that are commonly used in lieu of primary RASFs. In the current study, we investigated the effect of stimulation by tissue necrosis factor alpha (TNF-α) and interleukin 17 (IL-17) on primary and immortalized RASFs in order to gauge the appropriateness of the use of immortalized RASFs, the MH7A cell line, in the study of RA pathogenesis. Changes in the levels of secretion and expression of 8 proteins associated with RA upon stimulation were assessed by multiplex immunoassay. IL-17 stimulation had a minimal impact on protein secretion and expression for primary and immortalized cells. Basic fibroblast growth factor (FGF-2) was not detectable for the primary cells but was detectable for the immortalized cells. In contrast, monocyte chemoattractant protein 1 (MCP-1) was detectable for primary cells but was undetectable for immortalized cells. In general, protein expression and secretion by cells stimulated with TNF-α were significantly increased. For primary cells, several proteins were below the limit of detection for unstimulated cells and cells stimulated with IL-17, while levels for TNF-α-stimulated cells were within the detectable range. For the same proteins, expression was observed for immortalized cells, regardless of stimulation, suggestive of constitutive activation of the NF-κB signaling pathway. The current study therefore provides strong evidence that immortalized and primary RASFs differ in regard to protein expression and secretion and therefore may not be appropriate for use in the study of RA pathogenesis.


Subject(s)
Arthritis, Rheumatoid/pathology , Cell Transformation, Viral , Fibroblasts/virology , Simian virus 40 , Aged , Cell Culture Techniques , Cell Line, Transformed , Chemokine CCL2/analysis , Female , Fibroblast Growth Factor 2/analysis , Fibroblasts/drug effects , Fibroblasts/pathology , Humans , Interleukin-17/pharmacology , Middle Aged , NF-kappa B/metabolism , Signal Transduction/drug effects , Signal Transduction/physiology , Synovial Fluid/cytology , Synovial Fluid/virology , Tumor Necrosis Factor-alpha/pharmacology
11.
Pediatr Infect Dis J ; 30(11): 980-2, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21633319

ABSTRACT

Nonbacterial arthritis is a rare but well-recognized complication of acute varicella in children. Reported cases usually described monoarthritis of the knee that occurs at the onset of the rash or shortly after. Herein, we describe a case of arthritis of the hip that occurred in a 4-year-old girl 6 days before the onset of rash. The presence of varicella-zoster virus DNA in synovial fluid was confirmed by real-time polymerase chain reaction. A review of the literature reported 26 previous cases of aseptic arthritis due to varicella infection among children.


Subject(s)
Arthritis, Infectious/virology , Chickenpox/virology , DNA, Viral/analysis , Exanthema/virology , Herpesvirus 3, Human/physiology , Hip/virology , Synovial Fluid/virology , Arthritis, Infectious/complications , Arthritis, Infectious/diagnosis , Chickenpox/complications , Chickenpox/diagnosis , Child, Preschool , Exanthema/complications , Female , France , Hip/pathology , Humans , Real-Time Polymerase Chain Reaction , Synovial Fluid/chemistry
12.
Scand J Immunol ; 70(3): 295-9, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19703019

ABSTRACT

To study the viral loads of human endogenous retrovirus HERV-K (HML-2) type 1 and type 2 in rheumatoid arthritis (RA), we measured the viral loads of HERV-K (HML-2) type 1 and type 2 using nucleic acid sequence-based amplification (NASBA) technology. We analyzed plasma samples from RA patients (n = 79) and healthy volunteers (HV, n = 46) and synovial fluid samples from RA (n = 10) and osteoarthritis (OA, n = 10) patients. HERV-K type 1 and type 2 viruses were detected and quantified for the majority of plasma and synovial fluid samples from RA patients. HERV-K type 1 and type 2 viral loads were significantly elevated in RA patients compared with HV in plasma (P < 0.0001) and from RA patients compared with OA patients in synovial fluid (type 1: P = 0.0007; type 2: P = 0.023). Moreover, an association was observed between the HERV-K type 1 viral load in plasma and the disease activity in RA patients (RA patients with low activity versus high activity P = 0.0129; RA patients with intermediate activity versus high activity P = 0.037). Our findings showed that HERV-K (HML-2) viral load can be detected in plasma samples from RA patients, with higher levels observed for those with active disease. There was an association of HERV-K type 1 levels with the disease activity.


Subject(s)
Arthritis, Rheumatoid/virology , Endogenous Retroviruses/isolation & purification , Osteoarthritis/virology , Synovial Fluid/virology , Viral Load , Adult , Arthritis, Rheumatoid/blood , Female , Humans , Male , Middle Aged , Osteoarthritis/blood
13.
J Rheumatol ; 36(5): 893-7, 2009 May.
Article in English | MEDLINE | ID: mdl-19435971

ABSTRACT

OBJECTIVE: Rheumatoid arthritis (RA) is a chronic inflammatory polyarthritis; while the cause is unknown, it has been speculated that an infectious agent could be the trigger for the disease. Numerous attempts at isolating an agent have been unsuccessful. Our purpose was to identify a virus from diseased tissue from a patient with RA. METHODS: Diseased tissue taken at the time of knee replacement surgery from a patient with RA was inoculated into several cell lines and observed for cytopathic effect. Cells from the tissue were also grown as explants and were examined for viruses. Synovial fluid drawn 4 years prior to the surgery and frozen at -70 degrees C was also inoculated into cell lines. Following the development of a cytopathic effect and identification of the agent, sera from 50 patients with rheumatoid factor (RF)-negative RA were examined for IgM antibodies to the agent. RESULTS: After many inoculations and numerous subpassages, measles virus was identified in 6 cell lines inoculated with either the minced tissue or synovial fluid. Six cell lines co-cultivated with one or more of 9 explants also showed the presence of measles virus. Measles virus was confirmed by immunofluorescence and by neutralization. Eleven of 50 (22%) sera samples from patients with RF-negative RA had IgM antibodies to measles virus recombinant nucleoprotein. CONCLUSION: There is an association between measles virus and RA.


Subject(s)
Arthritis, Rheumatoid/virology , Knee Joint/virology , Measles virus/isolation & purification , Synovial Membrane/virology , Antibodies, Viral/analysis , Arthritis, Rheumatoid/immunology , Arthritis, Rheumatoid/pathology , Cell Line , Cytopathogenic Effect, Viral , Female , Humans , Immunoglobulin M/analysis , Knee Joint/pathology , Male , Measles virus/growth & development , Measles virus/immunology , Middle Aged , Neutralization Tests , Synovial Fluid/virology , Synovial Membrane/pathology
14.
Vet Microbiol ; 138(1-2): 156-62, 2009 Jul 02.
Article in English | MEDLINE | ID: mdl-19339126

ABSTRACT

Small ruminant lentiviruses (SRLVs) cause different clinical forms of disease in sheep and goats. So far in Spain, Maedi visna virus-like (MVV-like) sequences have been found in both species, and the arthritic SRLV disease has never been found in sheep until a recent outbreak. Knowing that arthritis is common in goats, it was of interest to determine if the genetic type of the virus involved in the sheep arthritis outbreak was caprine arthritis encephalitis virus-like (CAEV-like) rather than MVV-like. Alignment and phylogenetic analyses on nucleotide and deduced amino acid sequences from SRLV of this outbreak, allowed a B2 genetic subgroup assignment of these SRLV, compatible with a correspondence between the virus genetic type and the disease form. Furthermore, an isolate was obtained from the arthritic outbreak, its full genome was CAEV-like but the pol integrase region was MVV-like. Although its LTR lacked a U3 repeat sequence and had a deletion in the R region, which has been proposed to reduce viral replication rate, its phenotype in sheep skin fibroblast cultures was rapid/high, thus it appeared to have adapted to sheep cells. This outbreak study represents the first report on CAEV-like genetic findings and complete genome analysis among Spanish small ruminants.


Subject(s)
Arthritis, Infectious/veterinary , Disease Outbreaks/veterinary , Lentivirus Infections/veterinary , Lentiviruses, Ovine-Caprine/genetics , Sheep Diseases/virology , Animals , Arthritis, Infectious/genetics , Arthritis, Infectious/virology , Base Sequence , Choroid Plexus/virology , Cloning, Molecular , DNA, Viral/genetics , DNA, Viral/isolation & purification , Genome, Viral , Lentivirus Infections/epidemiology , Lentiviruses, Ovine-Caprine/classification , Lentiviruses, Ovine-Caprine/isolation & purification , Phylogeny , Polymerase Chain Reaction/methods , RNA, Viral/genetics , Sheep , Spain , Synovial Fluid/virology , Synovial Membrane/virology , Terminal Repeat Sequences/genetics , Visna-maedi virus/classification , Visna-maedi virus/genetics , Visna-maedi virus/isolation & purification
15.
Eur J Immunol ; 38(11): 3237-44, 2008 Nov.
Article in English | MEDLINE | ID: mdl-18991285

ABSTRACT

Viruses may be part of the pathogenesis of rheumatoid arthritis (RA). Double stranded RNA (dsRNA) is a prototypic viral conformation of nucleic acid that is highly arthritogenic in mice. Therefore, we developed an ELISA to detect dsRNA in sera and synovial fluids (SF) in RA patients and in osteoarthritic controls. The developed ELISA recognizes picogram levels of viral or synthetic dsRNA but shows no reactivity against DNA, synthetic ssRNA, or total RNA prepared from mammalian cells. Before analysis by ELISA, each sample was subjected to RNA precipitation. The RA patients had significantly higher levels of dsRNA than the osteoarthritis patients in SF and in sera. In 7 of 17 RA patients, EBV was present in SF and in all but one of these this was accompanied by the presence of dsRNA. No parvovirus, cytomegalovirus, or polyomavirus was detected. The anti-viral cytokine IFN-alpha was detected in SF in 10 of 21 RA patients, but in none of the osteoarthritis patients. Notably, RA patients with erosive disease course had significantly higher levels of dsRNA in SF than non-erosive patients, but no correlation between dsRNA levels and the presence of RF or levels of C-reactive protein, IL-6, or IFN-alpha was observed.


Subject(s)
Arthritis, Rheumatoid/etiology , RNA, Double-Stranded/analysis , Synovial Fluid/chemistry , Adult , Aged , Aged, 80 and over , Enzyme-Linked Immunosorbent Assay , Female , Humans , Interferon-alpha/analysis , Male , Middle Aged , Synovial Fluid/virology
16.
J Rheumatol ; 35(7): 1265-70, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18484700

ABSTRACT

OBJECTIVE: To determine the prevalence and clinical significance of human parvovirus B19 (B19) infection in patients with rheumatoid arthritis (RA). METHODS: One hundred patients with RA and 94 apparently healthy blood donor controls were enrolled for study. Plasma samples of patients and controls were examined for the presence of anti-B19-specific antibodies by ELISA. B19 DNA was detected in plasma and peripheral blood leukocyte (PBL) samples of all patients and controls as well as in synovial fluid cells of 38 RA patients by nested polymerase chain reaction. Disease activity and clinical manifestations were determined in RA patients with and without markers of B19 infection. RESULTS: IgM anti-B19-specific antibodies were detected in 24.0% of RA patients; B19 DNA was found in plasma and/or PBL, synovial fluid cells in 34.0% (34 patients); in 14.0% of the cases (14 patients) both markers were found. In blood donor controls, anti-B19 IgM antibodies were observed in 16.0% (15 donors) and B19 DNA in 6.4% (6 donors); all donors with detectable B19 genomic DNA were IgM-positive. The disease activity in patients with and without B19 infection was similar, while the frequency of clinical complications was significantly higher in the patients with anti-B19 IgM antibodies. Moreover, liver failure and sicca syndrome were observed in the viremic patients only. CONCLUSION: Our study confirms observations regarding a high prevalence of B19 DNA in patients with RA, and a possible role of this viral infection in the pathogenesis of RA.


Subject(s)
Antibodies, Viral/blood , Arthritis, Rheumatoid/virology , DNA, Viral/blood , Parvoviridae Infections/complications , Parvovirus B19, Human , Adult , Aged , Aged, 80 and over , Arthritis, Rheumatoid/complications , Arthritis, Rheumatoid/epidemiology , Case-Control Studies , Humans , Immunoglobulin G/blood , Immunoglobulin M/blood , Liver Failure/complications , Liver Failure/virology , Male , Middle Aged , Parvoviridae Infections/blood , Parvoviridae Infections/epidemiology , Parvovirus B19, Human/immunology , Parvovirus B19, Human/isolation & purification , Prevalence , Synovial Fluid/virology
17.
Diagn Microbiol Infect Dis ; 60(4): 381-5, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18096351

ABSTRACT

We aimed to determine the possible role of parvovirus B19 (PVB19) in the etiology of osteoarthritis. PVB19 DNA, anti-VP1 IgM and IgG, and interleukin IL-6 levels were also assayed in synovial fluids of 42 patients with osteoarthritis and 10 controls. PVB19 DNA was detected in 28 of 42 (66.66%) in patients and in 3 of 10 (30%) in controls. IgG and IgM response were detected in 21 of 42 (50.00%) and in 2 of 42 (4.76%) patients, respectively. IL-6 were positive in 15 of 42 (36%) patients and in 3 of 10 (30%) controls. All IgG (+) samples had PVB19 DNA (100%, P < 0.001). Eleven of 15 IL-6 (+) samples had PVB19 DNA (+) (73.33%, P < 0.05). Moreover, all IL-6 (+) samples (n = 5) in stage IV had PVB19 DNA (+) (100%, P < 0.001). We have detected a significant association between the stages of osteoarthritis and PVB19 DNA (P < 0.05). These findings support the presence of PVB19 acting as a transactivator of IL-6 expression as reported earlier. Our results also suggest that the higher stages of osteoarthritis might be related to the increased inflammation and cell damage on joint cartilage due to PVB19.


Subject(s)
Osteoarthritis/virology , Parvoviridae Infections/diagnosis , Parvovirus B19, Human/isolation & purification , Synovial Fluid/virology , Aged , Aged, 80 and over , Antibodies, Viral/analysis , DNA, Viral/analysis , Female , Humans , Immunoglobulin G/analysis , Immunoglobulin M/analysis , Interleukin-6/analysis , Male , Middle Aged , Severity of Illness Index , Synovial Fluid/chemistry
18.
J Virol Methods ; 137(1): 95-102, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16839616

ABSTRACT

Fibroblast like synoviocytes are the main resident cells in normal joints and are known to play a major role in the pathogenesis of rheumatoid arthritis. Efficient gene targeting of fibroblast like synoviocytes (FLS) is a major goal of current ex vivo gene therapy approaches for the treatment of rheumatoid arthritis. However, there is a need to improve viral systems capable of delivering genes to human rheumatoid fibroblasts and attempts have been made to develop a protocol for high efficiency, reproducible gene transfer using a replication-defective retrovirus vector. The effects of different experimental conditions were examined as well as those related to cellular and viral features on the efficiency of transducing the retroviral-driven expression of enhanced green fluorescent protein (EGFP) to FLS harvested from patients with rheumatoid arthritis. The optimal method established involved a double round of infection by centrifugation with a resting period of 4h between rounds. This approach led to the transduction of 30-70% of FLS obtained from nine patients with rheumatoid arthritis. Consistent transduction efficiencies were achieved in repeat assays such that it could be inferred that the variations observed were attributable to the specific characteristics of each cell line. This simple protocol renders a consistent and reproducible efficiency of rheumatoid fibroblast transduction and makes stable gene targeting using non-replicating retrovirus derived vectors an affordable option for the treatment of rheumatoid arthritis.


Subject(s)
Fibroblasts/virology , Genetic Therapy/methods , Retroviridae/genetics , Synovial Fluid/virology , Transduction, Genetic/methods , Arthritis, Rheumatoid , Cells, Cultured , Genes, Reporter , Green Fluorescent Proteins/analysis , Humans , Reproducibility of Results
19.
Lik Sprava ; (5-6): 28-32, 2005.
Article in Ukrainian | MEDLINE | ID: mdl-16396287

ABSTRACT

The study of persistence in mononuclear leukocytes (ML) of blood and synovial fluid of 218 patients with rheumatoid arthritis (RA) Cytornegalovirus (CMV), the 1-st and 2-nd types of Herpes virus simplex (VH), Epstain-Barr virus (VEB), Mycoplasma arthritidis (Ma), Mycoplasma fermentans (Mf), Ureaplasma urealiticum (U), Chlamidia trachomatis (Ct), viruses of Hepatitis B and C was carry out by direct and indirect immunofruorescence, immunoenzymatic analysis and polymerase chain reaction. An increased frequency of contamination of blood ML with infectious agents in patients with RA was established (57,4% compared with 16,7% in control group). The following infectious agents were revieled more frequently: in ML of blood and synovial fluid the Ma (relatively 20,5% and 15,9%), Mf (15,6% and 13,2%), Ct (18,4% and 13,2%), VH (27,1% and 10,5%), VEB (12,7% and 5,3%) and CMV (11,2% and 7,9%). Types of frequency dynamics of ML contamination with these infectious agents in different time phases of RA were determined.


Subject(s)
Arthritis, Rheumatoid/blood , Infections/microbiology , Leukocytes, Mononuclear/microbiology , Synovial Fluid/microbiology , Antibodies, Bacterial/analysis , Antigens, Bacterial/analysis , Antigens, Viral/analysis , Chlamydia trachomatis/immunology , Chlamydia trachomatis/isolation & purification , Cytomegalovirus/immunology , Cytomegalovirus/isolation & purification , DNA, Bacterial/analysis , DNA, Viral/analysis , Female , Fluorescent Antibody Technique, Indirect , Hepacivirus/immunology , Hepacivirus/isolation & purification , Hepatitis B virus/immunology , Hepatitis B virus/isolation & purification , Herpes Simplex/classification , Herpes Simplex/immunology , Herpesvirus 4, Human/immunology , Herpesvirus 4, Human/isolation & purification , Humans , Infections/complications , Infections/virology , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/virology , Male , Mycoplasma arthritidis/immunology , Mycoplasma arthritidis/isolation & purification , Mycoplasma fermentans/immunology , Mycoplasma fermentans/isolation & purification , Polymerase Chain Reaction , RNA, Viral/analysis , Retrospective Studies , Synovial Fluid/immunology , Synovial Fluid/virology , Ukraine/epidemiology , Ureaplasma urealyticum/immunology , Ureaplasma urealyticum/isolation & purification
20.
Rheumatol Int ; 26(1): 7-11, 2005 Nov.
Article in English | MEDLINE | ID: mdl-15322815

ABSTRACT

In order to evaluate the role of human parvovirus B19 in the etiopathogenesis of autoimmune diseases such as rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), synovial fluid and blood specimens were collected at 1-month intervals from 20 patients with early synovitis (ES) and 31 with RA. Blood specimens were also collected from 25 patients with SLE, 25 with osteoarthritis (OA) as the diseased control group, and 50 healthy blood donors (HBD) as the healthy control group. Detection of B19 IgM and B19 IgG were performed by enzyme-linked immunosorbent assay from serum specimens, and B19 DNA was detected by polymerase chain reaction from synovial fluid samples. B19 IgM, B19 IgG, and B19 DNA were found in the three patients of the ES group. Subsequently, two of them were diagnosed with RA and one with SLE. B19 DNA was also detected in the synovial fluid of eight patients in the RA group. Of them, all were positive for B19 IgG and half were positive for B19 IgM. B19 IgM was not detected in either of the control groups. To define the role of B19 in the etiopathogenesis and prognosis of undiagnosed arthritis and other chronic inflammatory diseases such as RA and SLE, we need broader serial and prospective studies based on clinical and laboratory collaboration. In conjunction with case reports, these studies would also serve to detect other possible factors in the etiopathogenesis of chronic inflammatory diseases.


Subject(s)
Antibodies, Viral/blood , Arthritis, Rheumatoid/virology , Parvoviridae Infections/complications , Parvovirus B19, Human/isolation & purification , Synovitis/virology , Adult , Arthritis, Rheumatoid/blood , Arthritis, Rheumatoid/pathology , DNA, Viral/analysis , Female , Humans , Immunoglobulin M/blood , Immunoglobulins/blood , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/pathology , Lupus Erythematosus, Systemic/virology , Male , Middle Aged , Osteoarthritis/blood , Osteoarthritis/pathology , Osteoarthritis/virology , Parvoviridae Infections/blood , Parvoviridae Infections/pathology , Parvovirus B19, Human/genetics , Parvovirus B19, Human/immunology , Synovial Fluid/chemistry , Synovial Fluid/cytology , Synovial Fluid/virology , Synovitis/blood , Synovitis/pathology
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