Production of dimer-specific and dengue virus group cross-reactive mouse monoclonal antibodies to the dengue 2 virus non-structural glycoprotein NS1

Falconar, Andrew K.; Young, Paul R.

doi:10.1099/0022-1317-72-4-961

Cited by 65 publications

(83 citation statements)

References 29 publications

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“…Immunoaffinity purification of the native dimeric intracellular (iNS1) and extracellular (eNS1) forms of the DENV-2 NS1 glycoprotein from infected mammalian cell culture supernatants was described previously (11). To purify the DENV-2 eNS1 glycoprotein, clarified cell culture supernatants of DENV-2 (strain NG-C)-infected Vero cells were made to 30 mM Tris-HCl (pH 7.2) and 0.02% (wt/vol) NaN 3 with protease inhibitors (PPB).…”

Section: Methodsmentioning

confidence: 99%

Monoclonal Antibodies That Bind to Common Epitopes on the Dengue Virus Type 2 Nonstructural-1 and Envelope Glycoproteins Display Weak Neutralizing Activity and Differentiated Responses to Virulent Strains: Implications for Pathogenesis and Vaccines

Falconar

2008

Clin Vaccine Immunol

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The abilities of monoclonal antibodies (MAbs) that bind to defined sequential epitopes on the dengue virus (DENV) nonstructural-1 (NS1) glycoproteins to cross-react with epitopes on the DENV envelope (E) glycoproteins were investigated. In this study, some of these MAbs cross-reacted with the DENV type 2 (DENV-2) E glycoprotein and with synthetic peptides representing X-ray crystallographically confirmed surface-exposed regions on this glycoprotein. MAb 1G5.3 cross-reacted with the flavivirus-conserved 101-WGNGCGLFG-109 fusion sequence, the 273-SSGNL-277 DENV-2 hinge region sequence, and the 156-GKHGKEIKIT-165 sequence of virulent DENV-2 strains. MAb 1G5.4-A1-C3 cross-reacted with the 67-NTTTESRCPT-76 and 156-GKHGKEIKIT-165 sequences of virulent DENV-2 strains, the 338-EIMDLDNRHV-347 sequence from a highly virulent DENV-2 (M2) strain, and two epitopes on a virulent DENV-3 strain (288-KMDKLELKG-296 and 323-RVEYRGEDAP-332), which all contained target ELK/KLE-type motifs (underlined). These MAbs showed reduced cross-reactions with the corresponding sequences from weakly pathogenic strains of all four DENV serotypes and had either no (MAb 1G5.4-A1-C3) or weak (MAb 1G5.3) neutralizing activity against them. MAb 1G5.3 more strongly neutralized DENV-2 strains with higher pathogenic capacities, while MAb 1G5.4-A1-C3 showed increasing neutralizing titers against the virulent DENV-3 strain and the moderately virulent and highly virulent (M2) DENV-2 strains. These cross-reactions with the E glycoprotein accord with the observation that MAb 1G5.3 caused dramatic and lethal antibody-enhanced replication (AER) of a DENV-2 strain in vivo. Together with in vivo AER studies of these DENV strains using MAb 1G5.4-A1-C3, these results may account for the increased pathogenic capacities of such strains, which is likely to have important implications for pathogenesis and vaccines.The spread of dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) throughout the world has occurred through transportation of the more virulent viral strains from Southeast Asia, where DHF/DSS is the main cause of juvenile hospitalization (14). Strains of dengue virus type 2 (DENV-2) and DENV-3 are associated with most cases of DHF/DSS, but there are no reliable virulence marker sequences on pathogenic DENV strains. Nearly all DHF/DSS cases result from sequential infection with a virulent DENV strain of another serotype after the initial infection (14, 15). Patient antibodies bind to common epitopes on the heterologous virus, and instead of cross-neutralization, they can enhance the replication of DENV strains in target Fc receptor-bearing monocytes/ macrophages, which has been hypothesized to account for DHF/DSS (15). The majority of evidence for antibody-enhanced replication (AER) of the DENVs comes from in vitro studies, but dramatic AER of a DENV-2 strain has also been demonstrated in vivo (10; see below).Human immunoglobulin G (IgG) polyclonal antibodies (PAbs) generated against the DENV nonstructural-1 (NS1) glycoprotein could be detected only dur...

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Section: Methodsmentioning

confidence: 99%

Monoclonal Antibodies That Bind to Common Epitopes on the Dengue Virus Type 2 Nonstructural-1 and Envelope Glycoproteins Display Weak Neutralizing Activity and Differentiated Responses to Virulent Strains: Implications for Pathogenesis and Vaccines

Falconar

2008

Clin Vaccine Immunol

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“…Virus-infected HMEC-1 monolayers grown on glass coverslips were fixed in 3?7 % formaldehyde for 20 min and were then permeabilized for 5 min with 0?05 % Triton X-100 in PBS. After rinsing with PBS, cells were blocked with PBS/2 % BSA for 30 min and exposed for 1 h at 37 uC to mAbs to D2V E protein (donated by the Pedro Kourí Institute, Havana, Cuba) or NS1 protein (donated by A. Falconar, London School of Hygiene & Tropical Medicine, U.K; Falconar & Young, 1991) at 1 : 10 and 1 : 50 dilutions, respectively. After washing with PBS, cells were incubated with FITC-conjugated anti-mouse IgG at 1 : 200 dilution (Molecular Probes) for 1 h at 37 uC.…”

Section: Methodsmentioning

confidence: 99%

IL8 release, tight junction and cytoskeleton dynamic reorganization conducive to permeability increase are induced by dengue virus infection of microvascular endothelial monolayers

Talavera

Castillo

Domı́nguez

et al. 2004

Journal of General Virology

183

152

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Permeability alterations of microvascular endothelia may be a factor in the plasma leakage produced by dengue virus infection. Confluent monolayers of the human dermal microvascular endothelial cell line HMEC-1 were utilized as an experimental model to study the cellular responses induced by the virus. Infected monolayers showed increased permeability for [ 3 H]mannitol, but no changes were observed for 4-70 kDa dextrans at 48 h post-infection (p.i.), a time at which viral titres reached maximal values and 40 % of the cells expressed viral proteins. A further increase in permeability occurred at 72 h, still without evident cytopathic effects on the monolayer. Coinciding with this, actin was reorganized in the infected cells and the tight junction protein occludin was displaced to the cytoplasm. Increments in the thickness of stress fibres and focal adhesions were observed in uninfected cells neighbouring infected cells. Culture medium from infected monolayers induced permeability changes and thickening of actin-containing structures in control cultures that resembled those observed 48 h p.i. Interleukin (IL) 8 was found in culture medium at concentrations ranging from 20 to 100 pg ml "1 . Neutralizing antibodies against IL8 partially inhibited the changes produced by the culture medium as well as those induced by addition of IL8. Genistein inhibited the effect of the culture medium and the phosphorylation of proteins associated with focal adhesions and indicated the participation of tyrosine kinases. These findings suggest that IL8 production by infected monolayers contributes to the virus-induced effect on the cytoskeleton and tight junctions and thereby modifies transendothelial permeability.

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“…Polyclonal and monoclonal antibodies to the NS1 protein have identified type-, complex-, and flavivirus-specific epitopes [34], which are mainly conformation dependant but are not virus neutralising. The NS1 protein of WNV has also been demonstrated to be antigenic and has been used as a diagnostic reagent in antibody capture ELISA procedures [44].…”

Section: The Envelope Protein Ementioning

confidence: 99%

West Nile virus infection of horses

2004

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-West Nile virus (WNV) is a flavivirus closely related to Japanese encephalitis and St. Louis encephalitis viruses that is primarily maintained in nature by transmission cycles between mosquitoes and birds. Occasionally, WNV infects and causes disease in other vertebrates, including humans and horses. West Nile virus has re-emerged as an important pathogen as several recent outbreaks of encephalomyelitis have been reported from different parts of Europe in addition to the large epidemic that has swept across North America. This review summarises the main features of WNV infection in the horse, with reference to complementary information from other species, highlighting the most recent scientific findings and identifying areas that require further research.

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Production of dimer-specific and dengue virus group cross-reactive mouse monoclonal antibodies to the dengue 2 virus non-structural glycoprotein NS1

Cited by 65 publications

References 29 publications

Monoclonal Antibodies That Bind to Common Epitopes on the Dengue Virus Type 2 Nonstructural-1 and Envelope Glycoproteins Display Weak Neutralizing Activity and Differentiated Responses to Virulent Strains: Implications for Pathogenesis and Vaccines

Monoclonal Antibodies That Bind to Common Epitopes on the Dengue Virus Type 2 Nonstructural-1 and Envelope Glycoproteins Display Weak Neutralizing Activity and Differentiated Responses to Virulent Strains: Implications for Pathogenesis and Vaccines

IL8 release, tight junction and cytoskeleton dynamic reorganization conducive to permeability increase are induced by dengue virus infection of microvascular endothelial monolayers

West Nile virus infection of horses

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