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1
Isa, Adiba. "Cellular immune responses against human parvovirus B19 infection /." Stockholm, 2006. http://diss.kib.ki.se/2006/91-7140-662-X/.
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Tolfvenstam, Thomas. "Human parvovirus B19 : studies on the pathogenesis of infection /." Stockholm, 2001. http://diss.kib.ki.se/2001/91-628-4710-4/.
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Norbeck, Oscar. "Clinical and immunological aspects of human parvovirus B19 infection /." Stockholm : Dept. of laboratory medicine, Karolinska institutet, 2005. http://diss.kib.ki.se/2005/91-7140-203-9/.
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Heegaard, Erik D. "Diagnostic and clinical aspects of human parvovirus B19 infection /." [S.l. : Erik D. Heegaard], 2003. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=010694314&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
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Blundell, Matthew Charles. "Characterization of the promoter in a human B19 parvovirus." Thesis, University of British Columbia, 1989. http://hdl.handle.net/2429/29011.
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The nucleotide sequence of the B19-Wi isolate of an autonomous human parvovirus was determined and compared with the sequence of the closely related isolate, B19-Au. The B19-Wi genome was similar to the B19-Au genome, as shown from DNA sequence analyses. It had been previously suggested from the sequence of the B19-Au genome, that the termini may be imperfect inverted terminal repeats. The additional sequence present on the right-hand terminus of B19-Wi supported that supposition. The hairpin termini of the B19 genome were of the same type as those found in adeno-associated parvoviruses, and suggested B19 may be more closely related to AAV viruses than to the autonomous parvoviruses.
In vitro transcription with HeLa nuclear extracts using B19-Wi DNA identified a single active RNA polymerase II promoter between map unit 5 and 6. A single promoter was unique; all other parvoviruses characterized to date have two or more active promoters. A series of ordered deletions, prepared in the region upstream of the initiation site of the promoter, indicated that multiple cis-activating motifs were required to maximize in vitro transcription.
A transcription factor in HeLa cells specifically bound to and protected a GC rich sequence or GC-box upstream of the promoter, as shown by gel-shift competition assays, and DNAse I and DMS footprinting assays. The protected GC-box had the consensus sequence of a high affinity binding site for the trans-activating factor SP1. The GC-box also formed the distal element of a tandem SPl-like motif, 21 bp upstream of the active TATA box. Synthetic oligonucleotides containing the GC-box specifically bound a HeLa factor and also depressed in vitro transcription from the B19 promoter when added as a competitor. Although not conclusive, binding of the HeLa factor may be inhibited by methylation of a cytosine residue within the GC-box. In vitro transcriptional activity decreased when the GC-box upstream of the B19 promoter was modified by site specific mutagenesis. Preliminary identification of other cis-activating motifs, which include two sequences recognized by factors that are functional both in transcription and replication, suggested that the B19 promoter is a complex regulatory region.Medicine, Faculty of
Biochemistry and Molecular Biology, Department of
Graduate
6
Morey, Adrienne Louise. "The pathogenesis of parvovirus B19 infection in the human fetus." Thesis, University of Oxford, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.316848.
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Carlsen, Karen Marie. "Human parvovirus B19 erythrovirus : methods established for virological and diagnostic aspects /." Copenhagen : Blackwell Munksgaard, 2006. http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&doc_number=014982739&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA.
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Lundqvist, Anders. "Clinical and laboratory findings in patients with persistent parvovirus 19 infection /." Stockholm, 2006. http://diss.kib.ki.se/2006/91-7140-828-2/.
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Sanchez, Jonathan L., Zachary Romero, Angelica Quinones, Kristiane R. Torgeson, and Nancy C. Horton. "DNA Binding and Cleavage by the Human Parvovirus B19 NS1 Nuclease Domain." AMER CHEMICAL SOC, 2016. http://hdl.handle.net/10150/622382.
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Infection with human parvovirus B19 (B19V) has been associated with a myriad of illnesses, including erythema infectiosum (Fifth disease), hydrops fetalis, arthropathy, hepatitis, and cardiomyopathy, and also possibly the triggering of any number of different autoimmune diseases. B19V NS1 is a multidomain protein that plays a critical role in viral replication, with predicted nuclease, helicase, and gene transactivation activities. Herein, we investigate the biochemical activities of the nuclease domain (residues 2-176) of B19V NS1 (NS1-nuc) in sequence-specific DNA binding of the viral origin of replication sequences, as well as those of promoter sequences, including the viral p6 and the human p21, TNF alpha, and IL-6 promoters previously identified in NS1-dependent transcriptional transactivation. NS1-nuc was found to bind with high cooperativity and with multiple (five to seven) copies to the NS1 binding elements (NSBE) found in the viral origin of replication and the overlapping viral p6 promoter DNA sequence. NS1-nuc was also found to bind cooperatively with at least three copies to the GC-rich Spl binding sites of the human p21 gene promoter. Only weak or nonspecific binding of NS1-nuc to the segments of the TNF alpha and IL-6 promoters was found. Cleavage of DNA by NS1-nuc occurred at the expected viral sequence (the terminal resolution site), but only in single-stranded DNA, and NS1-nuc was found to covalently attach to the 5' end of the DNA at the cleavage site. Off-target cleavage by NS1-nuc was also identified.
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Danober, Pascal Masutti Jean-Pierre. "Découverte d'un cas de sphérocytose héréditaire au décours d'une infection à Parvovirus B19 chez l'enfant à propos d'une observation /." [S.l] : [s.n], 2004. http://www.scd.uhp-nancy.fr/docnum/SCDMED_T_2004_DANOBER_PASCAL.pdf.
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Garcia, Sheila de Oliveira. "O significado das variantes do eritrovírus em pacientes com citopenias de origem desconhecida." Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/5/5164/tde-03112010-172833/.
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O eritrovírus humano (parvovírus), gênero Erytrovírus, é o único representante da família Parvoviridae responsável por um amplo espectro de doenças. Estudos recentes têm demonstrado variações entre o eritrovírus e orientam a reclassificação destas variantes em três genótipos distintos: genótipos 1, 2 e 3. O papel do eritrovírus na etiopatogenia de doenças hematológicas em humanos permanece incerto. Este estudo teve como objetivo principal avaliar a relação etiopatogênica dos genótipos do eritrovírus e as citopenias de origem desconhecida. Materiais e Métodos: Participaram do estudo 285 indivíduos procedentes do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo. Destes, 120 apresentavam citopenias de origem desconhecida (grupo 1 Casos), 45 eram doadores de medula óssea (grupo 2 Controles Saudáveis) e 120 eram pacientes com doenças oncohematológicas crônicas (grupo 3 Controles com Neoplasias Hematológicas). A pesquisa do vírus foi realizada pelo método de semi-nested PCR (Reação em Cadeia da Polimerase) em amostras de medula óssea e de sangue periférico. As fitas complementares foram seqüenciadas diretamente do produto da PCR. Amostras de plasma de todos os indivíduos incluídos no estudo foram testadas para presença de anticorpos IgG e IgM específicos contra o eritrovírus por ensaio imunoenzimático. Resultados: Dos 40 indivíduos com resultado positivo na PCR em amostra da medula óssea, o genótipo 1 foi encontrado em 22 (55%), o genótipo 2 em 5 (12,5%), o genótipo 3 em 13 (32,5%). Quando comparadas as freqüências de positividade entre os casos e controles (Grupo 1 VS Grupos 2 e 3), não encontramos diferença significativa com relação ao genótipo 1 (p=0, 192) nem com relação aos genótipos 2 e 3 (p= 0.143). A soroprevalência encontrada na amostra foi de 71%. Conclusão: Concluímos que a infecção isolada pelo eritrovírus, independente do genótipo encontrado, não tem relação etiopatogênica com as citopenias de origem desconhecida, uma vez que o vírus foi encontrado com a mesma freqüência nos casos e nos controles estudadosThe human erythrovirus (parvovirus), genus Erytrovirus, is the only representative of the family Parvoviridae responsible for a broad spectrum of diseases. Recent studies have shown variations within the erythrovirus and guide the classification of these variants in three distinct genotypes: genotypes 1, 2 and 3. The role of the erythrovirus in the etiopathogenesis of hematological diseases in humans remains uncertain. This studys main objective was to evaluate the etiopathogenic relationship between the genotypes of the erythrovirus and the cyptopenias of unknown origins. Methods and Materials: 285 individuals coming from the Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo participated in the study. Of these, 120 represented cytopenias of unknown origins (group one Cases), 45 were bone marrow donors (group two - Healthy Controls), and 120 were patients with chronic oncohematological diseases (group three Controls with Hematological Disorder). The research of the virus was done through the semi-nested PCR method (polymerase chain reaction) in bone marrow and peripheral blood samples. The complementary strands were sequenced directly from the product of the PCR. Plasma samples from all of the individuals included in the study were tested through immunosorbent assay for the presence of lgG and IgM antibodies specific to the eritrovírus. Results: Of the 40 individuals that had positive PCR bone marrow results, the genotype 1 was found in 22 (55%), the genotype 2 in 5 (12.5%), and genotype 3 in 13 (32.5%). When the frequency of positivity was compared between the cases and the controls (Group 1 vs. Groups 2 and 3), we did not find a significant difference in relation to genotype 1 (p=0.192), nor did we find a significant difference in relation to genotypes 2 and 3 (p=0.143). The overall seroprevalence found in the samples was 71%. Conclusion: We conclude that the infection isolated by the erytrovirus, independent of the genotype found, does not have a etiopathogenic relationship with the cytopenias of unknown origins, hence the virus was found with the same frequency in the cases and the controls studied
12
Chaput, CeÌcile Claire. "The establishment of an infectivity assay for human parvovirus B19 to investigate the efficacy of protocols for the inactivation of pathogens from plasma products." Thesis, Open University, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.434971.
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SALVETE, MARIE-JOSE. "Les infections a parvovirus humain b19 : une pathologie sous-estimee ?" Limoges, 1989. http://www.theses.fr/1989LIMO0133.
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Pallier-Kerestedjian, Coralie. "Etude de la permissivité cellulaire A : la réplication du parvovirus humain B19." Paris 5, 1994. http://www.theses.fr/1994PA05P160.
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Guiramand, Sonia. "Production de la protéine non structurale du parvovirus humain B19 en système procaryote." Paris 5, 1996. http://www.theses.fr/1996PA05P191.
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UNGIER, SERGE. "Madalie de minskowski-chauffard revelee par une erythroblastopenie aigue a parvovirus humain b19 : a propos de deux observations." Lille 2, 1992. http://www.theses.fr/1992LIL2M054.
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Pillet, Sylvie. "Synthèse et transport des protéines de capside du parvovirus humain B19 dans les cellules primaires érythroi͏̈des CD36+." Paris 7, 2002. http://www.theses.fr/2002PA077152.
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Sol, Nathalie. "Etude de l'interaction de la protéine NS1 du parvovirus humain B19 avec la région LTR du rétrovirus VIH1." Paris 5, 1992. http://www.theses.fr/1992PA05P004.
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Camus, Tobar Carolina. "Detección de Parvovirus B19 en muestras de pacientes con manifestaciones clínicas asociadas a la infección con el virus." Tesis, Universidad de Chile, 2011. http://repositorio.uchile.cl/handle/2250/131186.
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Memoria para optar al Título Profesional de Médico VeterinarioFundamentos: Parvovirus B19 (PV-B19) pertenece a la familia Parvoviridae, género Eritrovirus. Es un virus ADN de hebra simple que presenta secuencias palindrómicas en sus extremos. Es muy prevalente en la población general, llegando a detectarse anticuerpos anti PV-B19 en más del 85% de la población geriátrica. Este virus es el agente causal de una amplia gama de manifestaciones clínicas, cuya severidad depende del estado inmunológico y hematológico del hospedero, e incluye el eritema infeccioso, problemas hematológicos y reumatológicos e hidrops fetal, entre otras. Objetivo: Se determinó la presencia de Parvovirus B19 en pacientes con manifestaciones clínicas asociadas a este virus. Materiales y Métodos: Se analizaron 262 muestras de sangre de pacientes provenientes de distintos centros hospitalarios de la Región Metropolitana y del Servicio de Diagnóstico del Programa de Virología, Facultad de Medicina, Universidad de Chile. De estas muestras, 211 fueron analizadas mediante la técnica de ELISA para la pesquisa de IgM anti PV-B19, 244 con PCR anidado (PCR-Nested), para detectar genoma viral y 185 muestras con ambas técnicas. Resultados: De las muestras analizadas mediante la técnica de PCR anidado, se detectó genoma viral en un 25% de éstas y de las muestras analizadas mediante la técnica de ELISA, se localizó IgM anti PV-B19 en un 19%. Considerando sólo las 185 muestras que fueron analizadas con ambas técnicas, se detectó un 31.9% de positividad. En los casos positivos para el virus, las manifestaciones clínicas prevalentes fueron síndrome febril y eritema infeccioso
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Leruez-Ville, Marianne. "La proteine non structurale du parvovirus humain b19 : role dans l'infection semi-permissive, production par un clone cellulaire stable et interaction avec le promoteur (doctorat : microbiologie)." Paris 11, 1998. http://www.theses.fr/1998PA114832.
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Morinet, Frédéric. "Production des antigenes structuraux du parvovirus humain b19 par expression des proteines de capside dans escherichia coli; leur utilisation pour developper un test diagnostic." Paris 7, 1990. http://www.theses.fr/1990PA077067.
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Le diagnostic de la plupart des infections humaines a parvovirus b19 repose sur la mise en evidence d'igm specifiques. En l'absence de systeme de culture cellulaire de routine permettant de produire du virus b19, seuls les serums de sujets viremiques representent la source d'antigene utile au diagnostic serologique. Pour pallier a cet inconvenient, nous avons clone un fragment de 1,4 kbp d'adn b19 situe dans la portion droite du genome; celle-ci code pour les proteines de capside. Ce fragment de 1,4 kbp a ete exprime en systeme prokaryote et a permis l'obtention de proteine hybride comportant des epitopes du b19. Cette proteine de fusion a ete utilisee pour mettre au point un test de depistage des igm seriques
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N'Guyen, Yohan. "Détection moléculaire des formes complètes et tronquées en région 5’non codante des Entérovirus et conséquences sur la réponse inflammatoire chez des patients souffrant de myocardite ou de cardiomyopathie dilatée Virus detection and semiquantitation in explanted heart tissues of idiopathic dilated cardiomyopathy adult patients by use of PCR coupled with mass spectrometry analysis Enterovirus but not Parvovirus B19 is associated with idiopathic dilated cardiomyopathy and endomyocardial CD3, CD68, or HLA-DR expression Major Persistent 5' Terminally Deleted Coxsackievirus B3 Populations in Human Endomyocardial Tissues Enterovirus Persistence in Cardiac Cells of Patients With Idiopathic Dilated Cardiomyopathy Is Linked to 5' Terminal Genomic RNA-Deleted Viral Populations With Viral-Encoded Proteinase Activities." Thesis, Reims, 2019. http://www.theses.fr/2019REIMM203.
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Des ARN génomiques d’Enterovirus (EV) tronqués en région 5’ Non-Codante ont été détectés dans les tissus cardiaques de cas de myocardite aigue et de cardiomyopathie dilatée (CMD). La cinétique d’apparition de ces formes virales cardiaques et leurs conséquences sur la réponse inflammatoire sont inconnues. Une technique de PCR-MS a permis de détecter des ARN d’EV seuls (32%) ou associés à l’ADN du PVB19 (48%) chez des patients souffrant de CMD idiopathique. Chez ces patients, la présence exclusive d’ARN EV était associée avec un immunomarquage endomyocardique positif pour CD3, CD68 ou HLA-DR. Dans ces cas de CMD, une stratégie de « RACE-PCR » a montré que les populations EV tronquées de 37 à 50 nucleotides (nt) représentaient les formes persistantes majoritaires (80%) associées à des formes tronquées intermédiaires (19%) et complètes (1%). Dans des cas de myocardite à EVs, les proportions des ARN tronqués de 37 à 50 nt (84%) étaient supérieures à celles des formes intermédiaires et complètes (P<10-3). Dans le sous groupe des patients avec une myocardite sévère, les proportions des populations tronquées de 8 à 36 nt étaient supérieures (P=0.02) et associées à des niveaux plus élevés d’ARNm d’IFN-β (P=0.02). La transfection de cardiomyocytes par des ARN viraux synthétiques tronquées de 8 à 36 nt dans des proportions identiques à celles des cas sévères, induit des niveaux supérieurs d’ARNm (P<10-3) et protéiques d’IFN-β (P=0.02). Les populations EV tronquées en région 5‘NC sont majoritaires dès la phase de myocardite aigue et les proportions des formes minoritaires tronquées de 15 à 36 nt pourraient moduler l’activation de la voie des IFN-β et la sévérité de la pathologieMajor enterovirus (EV) populations characterized by 5’ terminal genomic RNA deletions (TD) ranging up to 50 nucleotides were previously identified in cardiac tissues from acute myocarditis and chronic dilated cardiomyopathy (DCM) patients. Dynamics of emergence and impact of various EV-TD populations onto the inflammatory response remains unknown. Using a PCR-MS approach EV-RNAs were detected alone (32%) or with PVB19 genomes in 48 % of patients with an idiopathic DCM. Among these patients, EV- RNA was associated with a positive endomyocardial CD3, CD68 ou HLA-DR immunostaining. In these EV-DCM cases, a quantitative "RACE-PCR" system showed that 37 to 50 nt EV-TD forms were the major persistant viral populations (80%) in association with 15 to 36 nt EV-TD (19%) and full-length (FL) (1%) forms. In samples from myocarditis cases, levels of 37 to 50 nt EVB-TD forms (84%) appeared to be statistically higher than other EV-TD (8%) and FL forms (8%) (P<10-3). Among severe myocarditis cases subgroup, levels of 15 to 36 nt EV-TD forms were significantly higher (P=0.02)) and associated with higher IFN-β mRNA levels (P=0.02)) than in non-severe myocarditis patients. HCM transfection of synthetic 8 to 36 nt EV-TD forms induced higher IFN-β mRNA (P<10-3) and cytokine levels (P=0.02) comparatively to those obtained after transfection by others deleted EV RNA forms. EV-RNA TD populations appeared to be major in acute myocarditis and DCM cases. Moreover, the proportions of minor 15 to 36nt EV-TD forms could modulate the innate immune sensing mechanisms in cardiomyocytes and therefore the clinical severity of cardiac infection
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Huai-Sheng and 許懷升. "Baculovirus Expression of Human Parvovirus B19 Structural Protein." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/46574514128498032351.
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碩士中山醫學大學
免疫學研究所
99
Previous studies have indicated that the capsids of human parvovirus B19 are composed of VP1 and VP2 proteins, also expression of VP2 alone in eukaryotic expression system could self-assemble into empty capsids, no matter in size, appearance, antigenicity or immunogenicity are similar to native virions. The empty capsids which without genetic materials are called virus-like particles (VLPs). We used Spodoptera frugiperda (Sf9) cells infected with recombinant baculovirus expressing B19 VP2 capsid proteins. Polymerase chain reaction, Western Blot and indirect immunofluorescence assay were used to identify the VP2 proteins in infected cells. We hope to employ the platform of VP2 proteins expression for B19 diagnosis and vaccine development in the future.
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Shih-jing and 施靖瑜. "The Effects Of Human Parvovirus B19 VP1Unique Region On Macrophage." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/67061420760538064278.
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碩士中山醫學大學
免疫學研究所
96
Human Parvovirus B19 (B19) have two structural protein ,VP1 (83KDa) and VP2 (58KDa), which are identical except for 227 amino acids at the amino-terminal end of the VP1-protein, the so-called VP1-unique region (VP1u). Recently, a phospholipase A2 (PLA2) motif was identified in the VP1u and has been classified as group XIII sPLA2, which the mechanism remains unclear. Previous studies have shown that B19-VP1u PLA2 motif can stimulate activation of synoviocytes. However, the effects of B19-VP1u PLA2 motif on initiating innate immunity by macrophages are still unknown. In this study, we found that the purified B19-VP1u proteins had sPLA2 activity but the purified B19-VP1uD175A proteins did not. We also found that B19-VP1u significantly induced mouse macrophage cells RAW264.7 migration and phagocytosis as compared to mutant VP1uD175A proteins and control. In addition, we found that VP1u-PLA2 motif induced the inflammatory cytokine expression, including IL-6, IL-1β, and GM-CSF by multiplex PCR. Moreover, we found that VP1u-PLA2 motif induced the MMP-9 expression. These findings may offer some clue in understanding the effects of macrophage on B19 infection and for vaccine development in the future.
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Hsu, Tsai-Ching, and 徐再靜. "The Study of Human Parvovirus B19 Infection And Autoimmune Diseases." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/96565108112425923911.
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博士中山醫學大學
醫學研究所
92
Previously, we described four patients who had clinical diagnosis of erythema infectiosum and presented with skin rash, polyarthralgia, and polyarthritis. Anticardiolipin antibody (aCL) and anti-neutrophil cytoplasmic antibody (ANCA) were also positive in these patients. These data indicate parvovirus B19 may be linked to the induction of an autoimmune response. In order to understand the relationship between B19 infection and autoimmune diseases, we then launch a series of research on them. Parvovirus B19 DNA was detected in 17 of 72 patients with SLE, but not in patients with other systemic rheumatic diseases. Of the 17 patients with B19 DNA, only one had B19 IgG antibody and two had B19 IgM antibodies. B19 DNA was found more commonly in sera from SLE patients without anti-B19 antibodies than in those with anti-B19 antibodies (P<0.05). B19 infection in patients with SLE may due to lack of anti-B19 antibodies because of either the immunocompromised nature of the host or the use of immunosuppressive drugs. There was a higher prevalence of hypocomplementaemia and RP in patients with parvovirus B19 viraemia than in those without parvovirus B19 viraemia. Moreover, our data herein revealed that human parvovirus B19 infection was frequently found in patients with chronic hepatitis B or hepatitis C infection. The occurrence of liver dysfunction was not affected by B19 infection in patients with HBV and HCV infection (p>0.05). All of the 20 serum samples with B19 DNA from patients with chronic HBV infection and all of the 12 serum samples with B19 DNA from patients with chronic HCV infection exhibited TW-3 genotype and TW-9 genotype, respectively. The frequency of B19 IgG(+)IgM(-) in HBV patients with B19 DNA PCR positivity was higher than in patients with PCR negativity (p<0.01). The results indicate that B19 may not be eradiated in these patients with HBV and HCV infection, causing persistent infection. Human parvovirus B19 has been found in various tissues in addition to erythroid lineage cells and nonstructural protein (NS1) is reported to induce cytotoxicity and apoptosis in erythroid lineage cells, but the mechanism in non-permissive cells is still not clear. So we attempted to elucidate the possible molecular mechanism of NS1-induced apoptosis in non-permissive cells and identified the expression of EGFP-NS1 fusion protein in COS-7 cells. The results suggest that the cell death of the NS1-transfected cells is associated with the mitochondria related apoptosis. The expression of p53, an important molecule in apoptosis and cell cycle regulation, and its downstream cell cycle kinase inhibitors p16INK4 and p21WAF1/CIP1 were up-regulated in the NS1-transfected cells. Also, increased expression of the pro-apoptotic Bcl-2 members Bax, Bad and activation of caspase 3 and caspase 9, but not the activation of caspase 8 nor Fas were detected in the NS1-transfected cells. Recently, the DNA array analysis revealed that p53, caspase 3 gene were up-regulation in NS-1 transfected cells. This is similar to our present findings. Moreover, we analyzed the CPR2 gene by using the RT-PCR method and demonstrated the difference was statistically significant between EGFP and EGFP-NS1 transfection in COS-7 cells (p<0.05). These findings might provide alternative information for further study in B19 infection and characterization of B19 NS1 protein in B19 non-permissive cells, and these will be used as reference to further study B19 pathogenesis.
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Ju, Cheng, and 鄭儒. "The study of human parvovirus B19 infection and anti-phospholipid antibodies." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/51656287136227222383.
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碩士中山醫學大學
生化微生物免疫研究所
104
Human parvovirus B19 (B19) is classified as Parvoviridae family and known causing disease in humans. The virus capsid is composed of two structural proteins, VP1 and VP2, which are identical except for 227 amino acids at the amino-terminal end of the VP1-protein, the so-called VP1-unique region (VP1u). B19-VP1u proteins have secretary phospholipase A2 (sPLA2) activity, which is essential for viral infectivity and as the region to neutralize the immune response (VP1u-IgG). Previous studies show B19-VP1u induces the production of anti-phospholipid antibody. To further understand the pathogenesis of B19- VP1u, we used different region of B19-VP1u recombinant proteins to analysis. The results indicated that (1) VP1u region of 61-227a.a. has higher sPLA2 activity; (2) 82-227a.a. region has higher association with anti-phospholipid antibody; (3) Passive immunization mice which injecting 31-227a.a. and 61-227a.a. antibodies may cause thrombocytopenia and aPTT prolong; (4) Passive immunization mice which injecting 21-227a.a and 51-227a.a antibodies may activate the macrophages migration; (5)The different VP1u region can stimulate CD8+/CD4+/Th17/Treg/NK/B cells expression. All together, we firstly demonstrated the region of B19-VP1u in B19 infection and the pathogenesis of anti-phospholipid antibodies in vivo.
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Van, Niekerk Albertus Bernhardus Willer. "Investigation of the role of human parvovirus B19 in chronic anaemia of HIV infected TB patients." Thesis, 1994. https://hdl.handle.net/10539/26637.
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A dissertation submitted to the Faculty of Medicine,
University of the Witwatersrand, in partial fulfilment
of the requirements for the degree
Master of Medicine (Virology)This study was undertaken to determine the role of human parvovlrus B19 (B19) in chronic anaemia of HIV infected TB patients. Patlents were selected from an existing databank of 307 patients included in a MRC HIV/TB study. Twenty-nine patients, 15 colnfected with HIV /TB and 14 Infected with TB only, were identified for further evaluation. These patient's era were subjected to serological and DNA detection studies using IgG and IgM ELISA methods and a nested polymerase chain reaction (PCR) assay. The selection of the nested PCR was based on comparative evaluation of a new rapid 99 cycle PCR method recommended for hepatitis B DNA detection and the nested PCR method established for B19. The nested assay was shown to be the more sensitive system in the context of B19 DNA detection. Serological evaluation of these 29 patients suggested that a greater proportion of HIV/TB patients with chronic anaemia had evidence of recent or past exposure to B19 than those not experiencing anaemia. The nested PCR demonstrated the presence of circulating B19 DNA in 2 coinfected individuals with haematological pictures compatible with persistent B19 infection. B19 DNA was also demonstrated in a TB only patient without anaemia; further haematological and serological evidence in this patient suggested recent exposure to B19. The serological and DNA amplification assay results of these 29 patients would suggest a possible role - either causal or co-factorial - for persistent B19 infection in the establishment of chronic anaemia in HIV/TB patients.
Andrew Chakane 2019
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Chun-Chou and 蔡鈞州. "The Study of Antibody Response to Human Parvovirus B19 Non-Structural Protein." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/5dyba5.
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碩士中山醫學大學
免疫學研究所
96
Human parvovirus B19 (B19) was classified as the Erythrovirus genus of the Parvoviridae family and the member known to be pathologic in hu-man. B19 is a single-stranded DNA virus with 5596 nucleotides, composed of capsid protein (VP) and non-structural (NS1) proteins. Previous studies have shown that the NS1-specific antibodies could be de-tected in B19 patients with chronic or persistent infection and associated with B19-related arthralgia. However, the role of anti-B19-NS1 antibody in patients with autoimmune diseases is still unknown. In this study nested PCR and ELISA were performed to determine the B19 diagnostic patterns by analyzing the serum samples of 95 patients with SLE, 98 patients with RA and 80 patients with clinical suspicion of B19 infection. Binding reactivity of B19-NS1 IgM and IgG antibodies were assessed by ELISA and Western blot. Our results shown that the prevalence of B19 IgM and IgG antibodies in B19 patients with recent infection [B19 IgM(+)IgG(-)DNA(-)] were significantly higher than other diagnostic patterns (P<0.001). In SLE, significantly higher prevalence of B19-NS1 IgM and IgG was observed in patients with recent infection [B19 IgM(+)IgG(-)DNA(-)], past infection [B19 IgM(-)IgG(+)DNA(-)] and persistent infection [B19 IgM(-)IgG(-)DNA(+)] rather than other diagnosis (P<0.05). In RA, significantly higher prevalence of B19-NS1 IgM and IgG was observed in patients with seronegative B19 diagnostic patterns [B19 IgM(-)IgG(-)DNA(-)] rather than other diagnosis (P<0.005). Altogether, the presence of both anti-B19-NS1 IgM and IgG antibodies could be an indicator in patients with recent B19 infection and B19-aasociated arthritis. Moreover IgM and IgG against B19-NS1 could be the biomarker on B19 infection and arthritis.
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Chen, Li-Jeng, and 陳俐礽. "Protective effects of Cystamine against human parvovirus B19-NS1 induced hepatic injury." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/62124371999683124828.
Full textAbstract:
碩士中山醫學大學
生化暨生物科技研究所
102
Human Parvovirus B19 (B19) infection is known to induce apoptosis and liver dysfunction. Cystamine is indicated in preventing apoptosis and it has been demonstrated to be beneficial on attenuating the hepatic injury in mice. Since our previous study demonstrated that B19-NS1 in development of autoimmunity by inducing apoptosis and aggravates liver injury, we herein intend to investigate the effect of cystamine on B19-NS1 induced hepatic iniury in BALB/c mice. For cystamine treatment, B19-NS1 BALB/c mice were injected intraperitoneally (i.p.) with cystamine daily for 14 days. Immunoblotting assay was performed to examine the signaling of inflammation in liver from B19-NS1 BALB/c mice given cystamine. Significant reduction of hepatic MMP-9 / MMP-2 activity ratio and decreased phosphorylation of p38, ERK, JNK, IKK-α, IκB and NF-κB (p-p65) proteins were observed in livers from B19-NS1 BALB/c mice given cystamine. Additionally, inflammation associated proteins, including MMP-9, CRP, IL-1β, IL-6 and TNF-α, were significantly decreased in the livers of B19-NS1 BALB/c mice given cystamine. Altogether, these results indicate that Cystamine has the protective effects against human parvovirus B19-NS1 induced hepatic injury.
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Tsai, Chun-Chou, and 蔡鈞州. "Study of Human Parvovirus B19 Proteins on Liver of NZB/W F1 mice." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/bkp37h.
Full textAbstract:
博士中山醫學大學
微生物免疫研究所
101
Human Parvovirus B19 (B19) infection has been associated with the production of various autoantibodies and recognized as a cause or trigger of autoimmune diseases. However, the precise mechanism is still unclear. Previous studies have reported that B19 may exacerbate or induce systemic lupus erythematous (SLE). Herein, we aimed to inves-tigate the effects of B19 on liver in NZB/W F1 mice by injecting B19 viral proteins. In Part I, we passive transfer of rabbit anti-B19-VP1u IgG was injected intravenously into NZB/W F1 mice. Significant increases of APhL, and anti-dsDNA antibody binding activity were detected in immunized anti-B19-VP1u mice. Additionally, significant increases of MMP9 activity and protein expression were detected in B19-VP1u IgG group. Notably, PI3K and phosphorylated ERK proteins were involved in the induction of MMP9. These results are indicated that the aggravated effects of anti-B19-VP1u antibody in disease activity of SLE. In part II, the recombinant B19 NS1, VP1u or VP2 proteins were injected subcutaneously into NZB/W F1 mice. Significant expressions of iNOS and COX2 proteins were detected and markedly hepatocyte disarray and lymphocyte infiltration were observed in livers from NZB/WF 1 mice receiving B19 NS1. Furthermore, significant increases of TNF-α, TNF-α receptor, IKK-α, IκB and NF-κB (p65) were detected in livers from NZB/W F1 mice re-ceiving B19 NS1. Accordingly, the MMP9 and uPA proteins were significantly increases in livers from receiving B19 NS1. These findings firstly demonstrated the aggravated effects of B19 NS1 but not VP1u or VP2 protein on hepatic injury and provide a clue in under-standing the role of B19 NS1 on hepatic injury in SLE.
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Chang, Shun-Chih, and 張舜智. "Study of Human Parvovirus B19-VP1u Proteins on Liver of Balb/c Mice." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/88115625755768995675.
Full textAbstract:
碩士中山醫學大學
微生物免疫研究所
101
Human Parvovirus B19 is a significant human pathogen in Parvoviridae. It has been re-ported that B19 can be detected in blood or liver specimens in patient with liver diseases. Indeed, various studies have postulated a connection between B19 infection and liver injury. Recently, studies suggested that the PLA2 motif of B19-VP1u play an important role in closely associated with B19-infected and inflammatory. However, the precise mechanism is still obscure. In the present study, we aimed to investigate the influence of B19-VP1u in Balb/c mice by injected subcutaneously with COS-7 cells expressing VP1u and observed using a non-invasive IVIS bio-luminescent imaging system. Our experimental results revealed that enhanced MMP-9 activity and increased phosphorylation of p38, ERK, IKK-α, IκB and NF-κB(p65) proteins in liver from Balb/c mice receiving B19-VP1u. Additionally, significantly increased inflammatory associated proteins, including CRP, IL-1β and IL-6 were observed in liver from Balb/c mice receiving B19-VP1u. Notably, phosphorylated STAT1 proteins were involved in the induction of inflammatory cytokines in liver from Balb/c mice receiving B19-VP1u. Our findings revealed the effects of B19-VP1u on liver injury and provide a clue in understanding the role of immuno-pathogenesis of B19-VP1u.
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St, Amand Janet Lynn. "Identification and characterization of small rnas and proteins expressed by the human parvovirus B19." Thesis, 1992. http://hdl.handle.net/2429/3329.
Full textAbstract:
The human pathogenic parvovirus B19 has a strict tissue tropism and
will only replicate in a subset of erythroid progenitor cells. However, it is
shown that COS-7 cells transfected with SV4O-B19 hybrid vectors express the
major B19 RNAs and proteins. In addition, capsid proteins synthesized in
these cells self-assemble into virus particles that by EM are morphologically
very similar to native B19 virions. Cytoplasmic RNA from transfected COS-7
cells was used to prepare a cDNA library using a method which enriched the
library for B19 cDNAs. A second cDNA library was prepared from B19 infected
human bone marrow cells that had been isolated from a patient with a
chronic myelogenous leukemia using PCR to amplify B19-specific cDNAs.
The libraries were probed with a series of RNA probes derived from
different regions of the B19 genome in pYT1O3 and selected cDNAs were
sequenced and compared. Two size classes of small, abundant, polyadenylated
RNAs were identified; the 700 and 800 nt size class of RNA is the product of
transcriptional processing in the middle of the B19 genome downstream from
an unusual polyadenylation signal, ATTAAA or AATAAC (map unit 49).
These transcripts contain an ORF within their second exon which is the same
reading frame used for the translation of the nonstructural proteins;
however, there are no AUG translation initiation codons within this region
of the ORF. The second most abundant size class of RNA in B19 infected cells
are the 500 and 600 nt transcripts which are made from three exons and
terminated at a normal polyadenylation signal (AATAAA) near the right
hand end of the genome (map unit 97). A 94 aa ORF within the third exon is
invariant in the two RNA species. [more abstract]
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Chun-Ching and 邱駿清. "The Effects of Antibody Against Human Parvovirus B19 VP1 unique region On Vascular Endothelial Cells." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/57807008492890659014.
Full textAbstract:
碩士中山醫學大學
免疫學研究所
98
Human parvovirus B19 infection has been frequently described as a cause or trigger of various autoimmune diseases. In previous studies, we have postulated the association among human parvovirus B19 (B19)-VP1unique region (VP1u), production of anti-beta2-glycoprotein I (anti-β2GPI) antibody and anti-phospholipid syndrome (APS)-like autoimmunity. However, the precise role of B19-VP1u in induction of APS is still obscure. To further elucidate the pathogenic roles of VP1u in B19 infection and autoimmunity, we examined the effect of anti-B19-VP1u IgG antibodies on endothelial cells that is recognized to play crucial roles in APS. Human vascular endothelial cells, ECV-304, were incubated with various preparations of purified human or rabbit IgG. The activation of endothelial cells and production of cytokines were assessed by flow cytometry and ELISA, respectively. Purified IgG from rabbits immunized with recombinant B19-VP1u proteins can up-regulate ICAM-1 (CD54), VCAM-1 (CD106), E-selectin (CD62E), MHC class II (HLA-DR, DP, DQ) molecule expression, and TNF-α production in endothelial cells as compared to those endothelial cells cultured with control IgG. These experimental results imply that antibodies against B19-VP1u play important roles in the immunopathological processes as well as human anti-β2GPI IgG that leads to development of APS. It could provide a clue in understanding the role of anti-B19-VP1u antibodies in APS manifestations.
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Huang, Zi-Yun, and 黃子芸. "The Effects of Human parvovirus B19 VP1 unique region on heart of Balb/c mice." Thesis, 2018. http://ndltd.ncl.edu.tw/handle/cfd78t.
Full text
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Tsung-Min and 林宗旻. "The Effects of Human parvovirus B19 VP1 unique region on heart of NZB/W F1 mice." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/58300165477411200242.
Full textAbstract:
碩士中山醫學大學
免疫學研究所
98
Human parvovirus B19 is one of the Parvoviridae family and known to be pathologic in humans. The capsid of parvovirus B19 consists of two structural proteins, VP1 and VP2, which are identical except for 227 amino acids at the amino-terminal end of the VP1 protein. So the additional 227 amino acids at VP1 amino-terminus were called VP1-unique region (VP1u). Recently, more and more literatures assumed that there is a link between B19 infection and myocarditis, but the detailed mechanisms are still unknown. In addition, parvovirus B19 infection has been associated with autoimmune diseases including SLE. Some studies have supposed that B19 may exacerbate or even induce SLE. In recent research, B19 VP1u was discovered that the region played an important role in successful infection and inflammation response. However, fewer literatures discussed whether B19 VP1u could cause myocarditis. In this study, we used NZB/W F1 mice as a lupus-prone model and inoculated these mice with (1) PBS, (2) Rabbit IgG and (3) Rabbit anit-B19 VP1u IgG respectively. We found that NZB/W F1 mice inoculated with B19-VP1u IgG had higher Aspartate Aminotransferase level in sera. Besides, we found that B19-VP1u IgG promoted protein expression and activity of matrix metalloproteinase 9 in heart via NFκB pathway. Moreover, B19 VP1u IgG didn’t cause myocardial infarction or hypertrophy. All together, we supposed that B19-VP1u IgG inoculation might directly or indirectly cause mice myocardial damage. In conclusion, anti-B19 VP1u IgG inoculation not only aggravates disease activity of SLE, but also causes cardiac damage and inflammation.
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Lee, Wen Chi, and 李汶錡. "The Effects of Human parvovirus B19 VP1 unique region antibody on heart of Balb/c mice." Thesis, 2018. http://ndltd.ncl.edu.tw/handle/geq8dh.
Full text
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van, Niekerk Albetus Bernhardus Willer. "Investigation of the role of human parvovirus B19 in chronic anaemia of hiv infected TB patients." Thesis, 1994. https://hdl.handle.net/10539/26457.
Full textAbstract:
A dissertation submitted to the Faculty of Medicine, University of the Witwatersrand, in partial fulfilment of the requirements for the degree Master of Medicine (Virology)This study was undertaken to determine the role of human parvovirus B19 (B19) in chronic anaemia of HIV infected TB patients. Patients were selected from an existing databank of 307 patients included in a MRC HIV/TB study. Twenty-nine patients, 15 coinfected with HIV/TB and 14 infected with TB only, were identified for further evaluation. These patients’ sera were subjected to serological and DNA detection studies using IgG and IgM ELISA methods and a nested polymerase chain reaction (PCR) assay.
IT2019
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Chen, Ren-Mo, and 陳壬模. "The study of human parvovirus B19 non-structure protein (NS-1) and 24-peptide antibodies in autoimmune diseases." Thesis, 2000. http://ndltd.ncl.edu.tw/handle/38042705581468569389.
Full textAbstract:
碩士中山醫學院
生物化學研究所
88
Objective: To analyze the reactivity of the autoimmune diseases patients sera with the parvovirus B19 proteins, 24- peptide and NS-1 was used as solid-phase antigen in an enzyme linked immunosorbent assay (ELISA) to screen human sera. Method : Sera from 80 patients with systemic lupus erythematous (SLE), 79 patientswith rheumatoid arthritis (RA), 4 patients with Sjogren''s syndrome (SS), 3 patients with systemic scleroderma (SSc), 4 patients with ankylosing spinitis (AS), 9 patients with uremia, All the patients’ sera were tested for B19 infection by using ELISA. NS-1 and 24-peptide were used to immunize rabbits. Sera from immunized rabbits were tested for B19 infection using antigens of NS-1, 24-peptide and proteinase 3 (PR3) , myeloperoxidases (MPO), E3, cardiolipin, ssDNA antigens by ELISA. Furthermore, anti-NS-1 antibodies were preincubated with different amounts NS-1 or E3 antigens, after antigens-antibodies absorption, anti-NS-1 antibodies were tested the direct and comptitive ELISA for antibody activity against E3, PR3, cardiolipin, ssDNA and MPO antigens. Results : The IgM and IgG antibodies against NS-1 and VP-1 peptide were higher in patients with RA than SLE . For example, Anti-NS-1 IgM and IgG antibodies were 31.5% and 35.2% in RA. 13.1% and 15.2% for SLE. For parvoscreen test which used VP-1 peptides as the antigens, 27.1% and 47.9% in RA patients. 4.3% and 30.4% for SLE patients. There is no difference between these two tests as determined by Chi-Square method. Sera from rabbits immunized with NS-1 antigens, recognize MPO, cardiolipin, E3, PR3, ssDNA and NS-1 antigens. But the binding of E3 and NS-1 antigens to anti-NS-1 antibodies was not affected after the antibody was absorpted by E3 antigens. Conclusion: The prevalence of anti-NS-1 IgM and anti-NS-1 IgG in patients with RA is higher than those in SLE, which is probably due to the higher infection rate of B19 virus in elderly people. Thus, the NS-1 antigens could be used to screen parvovirus B19 infection. Sera from NS-1 immunized animal can recognize E3, PR3, but these reaction could not be inhibited by E3 , suggesting that a cross-raction between the antibodies and the solid-phase antigens may occur.
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Lee, Yuan-Ming, and 李元明. "The Study of Serological and Molecular Epidemiology of Human Herpes Virus Type 8, Human Parvovirus Type B19 and Human Influenza Virus Type B Infection in Different Populations in Taiwan." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/68196045473080802153.
Full textAbstract:
博士國立陽明大學
公共衛生研究所
101
The aim of this study was to conduct serological and molecular epidemiology of Human herpes virus type 8( HHV-8), human parvovirus type B19( B19) and human influenza virus type B( Flu B) infections in different populations in Taiwan. HHV-8, the causal agent of Kaposi’s sarcoma, is transmitted sexually among homosexual men, but little is known of its transmission among injection drug users (IDUs). In contrast, B19, a causative agent for anemia, is most frequently detected in IDUs. A total of 744 serum samples among them 515 from IDUs and homosexual men attending a sexually transmitted disease (STD) clinic or from 229 HIV-based hospital unit were analyzed for anti-HHV-8 and anti-B19 antibodies. The presence of antibodies against HHV-8 and B19 were tested with ELISA, indirect immunofluorescence assay (IFA) and immunoblot assay (IBA). HHV-8 viral load was analyzed using a K6 gene real-time PCR assay. We used K1 gene nested-polymerase chain reaction (N-PCR) assay to amplify DNA of HHV-8 in HIV-1/AIDS patients. Furthermore, the DNA sequences were analyzed to determine inter-patient specific mutations and subtypes of HHV-8. The results showed that, among HIV-1/AIDS patients, men who have sex with men (MSM) had significantly higher HHV-8 infection rate (32.8% vs. 5.5%, P<0.0001) and significantly lower B19 infection rate (35.20% vs. 82.03%, P<0.001) than IDUs. The CD4 counts were not significantly different between HIV-1/AIDS patients with and without HHV-8. Phylogenetic analysis of 23 HHV-8 strains showed that there were 2 (8.7%)subgroup A, 17 (74%) subgroup C, 2 (8.7%) subtype E and 2 (8.7%) subgroup D. In summary, MSM had significantly higher prevalence rate of HHV-8 than IDUs in Taiwan. The independent association of HHV-8 infection with IDUs suggests that HHV-8 is transmitted through needle sharing, albeit less efficiently than B19. The results of this study are useful for designing preventive medicine and intervention strategies of those infectious diseases. Concerning Flu B study, the antigenic and genetic character of the virus was analyzed in patients registered to a sentinel surveillance site located in the northern region of Taiwan from 2006 to 2007. Both hemagglutinin (HA) test and fluorescence antibody assay were used to determine the isolation rate of Flu B in MDCK cells. Besides, antigenic analysis of HA in circulating Flu B was performed using hemagglutinin inhibition (HI) test. Finally, the phylogentic analysis and gene variation of Flu B were analyzed using one-step reverse transcription (RT)-PCR assay. The results showed that 143 of 452 clinical respiratory samples (32 %) were positive for B/Malaysia /2506/04-like strain ( It was recognized as a B reassortant lineage). Finally, we have confirmed the molecular evolution of Flu B gene and prospects for serum antibody responses after intradermal vaccination against virus. In the summary, The goal of this study is to determine the antigenic and genetic characterization of the hemagglutinins of influenza B viruses in Taiwan. Further investigation may be beneficial to improve strategies for the treatment and disease control of influenza B virus, and also helpful to the development of antiviral drug and vaccine.
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Chiu, Chun-Ching, and 邱駿清. "I. Mechanisms of the Beneficial Effects of Ocimum gratissimum aqueous extract on rats with CCl4-induced acute liver injuryII. Effects of Human Parvovirus B19 And Bocavirus VP1 Unique Region On Tight Junction Of Human Airway Epithelial A549 Cells." Thesis, 2015. http://ndltd.ncl.edu.tw/handle/04185308548567307464.
Full textAbstract:
博士中山醫學大學
微生物免疫研究所
103
Ocimum gratissimum (OG) is known as a food spice and traditional herb, which has been recommended for the treatment of various diseases. To investigate the hepatoprotective effect of OG aqueous extract (OGAE), male Wistar rats challenged by carbon tetrachloride (CCl4) were used as the animal model of hepatic injury. Our data imply that OGAE can effi-ciently inhibit CCl4-induced liver injuries in rats and may therefore be a potential food or herb for preventing liver injuries. As is widely recognized, human parvovirus B19 (B19) and human bocavirus (HBoV) are important human pathogens. Obviously, both VP1 unique region (VP1u) of B19 and HBoV exhibit the secreted phospholi-pase A2 (sPLA2)-like enzymatic activity and are recognized to participate in the pathogenesis of lower respiratory tract illnesses. However exactly how, both VP1u from B19 and HBoV affect tight junction has seldom been addressed. Therefore, this study investigates how B19-VP1u and HBoV-VP1u may affect the tight junction of the airway epithelial A549 cells by examining phospholipase A2 activity and transepithelial electrical resistance (TEER) as well as performing immunoblotting analyses. The results suggest that HBoV-VP1u rather than B19 VP1u likely plays more important roles in the disruption of tight junction in the airway tract. Meanwhile, this discrepancy appears not to be associated with the se-creted phospholipase A2 (sPLA2)-like enzymatic activity.