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At least 199 records · Page 11Linked to original sources
PubMed2002

Facilitation of cell adhesion by immobilized dengue viral nonstructural protein 1 (NS1): arginine-glycine-aspartic acid structural mimicry within the dengue viral NS1 antigen.

Dengue virus infection causes life-threatening hemorrhagic fever. Increasing evidence implies that dengue viral nonstructural protein 1 (NS1) exhibits a tendency to elicit potentially hazardous autoantibodies, which show a wide spectrum of specificity against extracellular matrix and platelet antigens. How NS1 elicits autoantibodies remains unclear. To address the hypothesis that NS1 and matrix proteins may have structural and functional similarity, cell-matrix and cell-NS1 interactions were evaluated using a cell-adhesion assay. The present study showed that dengue NS1 immobilized on coverslips resulted in more cell adhesion than did the control proteins. This cell adhesion was inhibited by peptides containing arginine-glycine-aspartic acid (RGD), a motif important for integrin-mediated cell adhesion. In addition, anti-NS1 antibodies blocked RGD-mediated cell adhesion. Although there is no RGD motif in the NS1 protein sequence, these data indicate that RGD structural mimicry exists within the NS1 antigen.

Amino Acid Sequence↗
PubMed1987

Dengue 4 virus monoclonal antibodies identify epitopes that mediate immune infection enhancement of dengue 2 viruses.

Nineteen monoclonal antibodies produced to dengue type 4 virus (DEN-4) strain 4328-S were tested for their ability to mediate antibody-dependent infection enhancement (ADE) with seven DEN-2 strains in P-388D1 mouse macrophage-like cells. In this first study of the distribution of enhancing epitopes on multiple DEN-2 strains reacted with monoclonal antibodies to a different serotype (DEN-4), DEN-4 monospecific antibodies produced ADE with DEN-2 viruses, indicating the presence of DEN-4-like determinants on DEN-2 viruses. Analysis differentiated at least one and possibly more DEN-2 strain subgroups, one of which (isolates AHF-110 and AHF-191) was previously identified by DEN-2 monoclonal antibody analysis. The study demonstrates the heterogeneous distribution of dengue complex and DEN-4 epitopes on DEN-2 strains. Monoclonal antibodies are valuable tools for study of the biology of ADE and its relation to dengue shock syndrome.

Animals↗
PubMedDate not supplied

The absence of dengue virus in the skin lesions of dengue fever.

Patients with confirmed dengue infection were studied in an attempt to elucidate the cause of the eruption of dengue fever. Punch biopsies of lesional skin were obtained and processed for the presence of viral antigen and immune globulins by the direct fluorescent antibody test. Another biopsy specimen and the acute sera were processed for virus isolation. Although virus was isolated from the serum of two patients, isolation attempts from the biopsy specimens of all four patients were negative. Fluorescent antibody studies revealed no evidence of either viral antigen or immune globulins in lesional skin. Although limited, the data suggest that the maculopapular rash of dengue fever may be caused by some mechanism other than direct viral infection of the skin or involvement of immune globulins.

Adolescent↗
PubMed1973

Dengue virus infection of mice: morphology and morphogenesis of dengue type-2 virus in suckling mouse neurones.

In dengue type-2 virus-infected neurones of suckling mice, formation of single-membrane vesicles is observed in the distended cisternae of the endoplasmic reticulum mostly of the perinuclear zone around 72 h after inoculation. Electron-dense 50-nm virus particles are arranged in chains in these distended cisternae; some form small crystalloid aggregates. Aberrant particles of different shapes are also seen in the distended cisternae about the same time that the virus particles appear. Parallel filamentous structures are occasionally observed in the cisternae that contain very few virions, either characteristic or aberrant. Increasing cytopathic changes are present after 75 to 96 h. There is an intense vesicular formation. Large numbers of virions and aberrant particles are seen either in the endoplasmic reticulum cisternae or smooth membrane vesicles. They are spread throughout the neurocytoplasm, extending into the dendrites. Dengue virions which are enclosed in fairly intact membrane-bound vesicles are released during cytolysis of the neurones. Morphogenesis of dengue virus type 2 is discussed.

Animals↗
PubMed2003

Dengue 2 PDK-53 virus as a chimeric carrier for tetravalent dengue vaccine development.

Attenuation markers of the candidate dengue 2 (D2) PDK-53 vaccine virus are encoded by mutations that reside outside of the structural gene region of the genome. We engineered nine dengue virus chimeras containing the premembrane (prM) and envelope (E) genes of wild-type D1 16007, D3 16562, or D4 1036 virus within the genetic backgrounds of wild-type D2 16681 virus and the two genetic variants (PDK53-E and PDK53-V) of the D2 PDK-53 vaccine virus. Expression of the heterologous prM-E genes in the genetic backgrounds of the two D2 PDK-53 variants, but not that of wild-type D2 16681 virus, resulted in chimeric viruses that retained PDK-53 characteristic phenotypic markers of attenuation, including small plaque size and temperature sensitivity in LLC-MK(2) cells, limited replication in C6/36 cells, and lack of neurovirulence in newborn ICR mice. Chimeric D2/1, D2/3, and D2/4 viruses replicated efficiently in Vero cells and were immunogenic in AG129 mice. Chimeric D2/1 viruses protected adult AG129 mice against lethal D1 virus challenge. Two tetravalent virus formulations, comprised of either PDK53-E- or PDK53-V-vectored viruses, elicited neutralizing antibody titers in mice against all four dengue serotypes. These antibody titers were similar to the titers elicited by monovalent immunizations, suggesting that viral interference did not occur in recipients of the tetravalent formulations. The results of this study demonstrate that the unique attenuation loci of D2 PDK-53 virus make it an attractive vector for the development of live attenuated flavivirus vaccines.

Animals↗
PubMed1978

A rapid fluorescent focus-inhibition test for determining dengue neutralizing antibody and for identifying prototype dengue viruses.

Neutralizing antibody to dengue virus in human and animal sera was assayed by the rapid fluorescent focus-inhibition test (RFFIT). Neutralizing-antibody titers could be detected after only 24 h compared to 5--6 days required by the plaque-reduction test. The RFFIT is more definitive than the conventional complement fixation (CF) or hemagglutination-inhibition (HI) test in identifying prototype dengue viruses, is reproducible, and is applicable to the routine detection of neutralizing antibodies to dengue viruses.

Antibodies, Viral↗
PubMedDate not supplied

Secondary dengue infection in schoolchildren in a dengue endemic area in the state of Rio de Janeiro, Brazil.

A seroepidemiologic survey was carried out in schoolchildren from public schools of the Niterói municipality, state of Rio de Janeiro, Brazil, after a period of sequential epidemics by dengue virus type 1 and 2 (DEN-1 and DEN-2). 450 blood samples were obtained by fingertip puncture and collected on filter paper discs. The hemagglutination inhibition (HAI) test was carried out using DEN-1 and DEN-2 antigens. HAI titres were demonstrated in 66% (297/450) of the sera and the geometric means of the titres were 1/182 and 1/71 for DEN-1 and DEN-2, respectively. Secondary infections were observed in 61% (181/297) of positive cases. Among these, 75% (135/181) were under fifteen years old. No dengue haemorrhagic fever (DHF) was reported in these children. Asymptomatic or oligosymptomatic infections were detected in 56% of the studied population. The absolute and relative frequencies of positive tests by age group and sex did not evidence statistically significant difference. The number of individuals infected probably produced a immunologic barrier responsible for the non occurrence of dengue epidemic in the latter years.

Adolescent↗
PubMed1991

Growth of dengue type 2 virus isolates in human peripheral blood leukocytes correlates with severe and mild dengue disease.

We tested three dengue type 2 (DEN-2) isolates from children with clinically apparent but mild secondary dengue infections, and 10 isolates from children with moderately severe dengue hemorrhagic fever, and noted significant growth differences in peripheral blood leukocytes, but not in C6/36 cells. We also observed cytopathic effects in C6/36 cells that correlated with disease severity. These preliminary observations suggest the possibility that viral factors, whether surface antigens, attachment sites for entry into leukocytes, or intrinsic replication properties in human mononuclear phagocytes, might contribute to enhanced DEN infection and to the severity of the disease.

Child↗
PubMed1996

Dengue-2 virus infection of human bone marrow: characterization of dengue-2 antigen-positive stromal cells.

Dengue is often associated with neutropenia and thrombocytopenia, suggesting that cells of the bone marrow may be targets of dengue viral infections. In this study we infected long-term marrow cultures with dengue type-2 (DEN-2) virus and characterized the viral antigen-positive cells. Using immunofluorescence microscopy and immunohistochemical staining, we demonstrated two types of stromal cells that were positive for DEN-2 virus antigens. The first was a population of relatively small (approximately 25 microns) CD11b/CD18 (MAC-1)-positive cells. When stained with anti-DEN-2 polyclonal antibody, these cells showed viral antigen-positive inclusions and, when stained with anti-tubulin or anti-vimentin antibodies, they showed a diffuse pattern of fluorescence, consistent with mobile dendritic cells with phagocytic functions. The second population of DEN-2 antigen-positive cells comprised a smaller proportion of the total cells. It was made up of larger cells (> 100 microns) that had a well-formed cytoskeletal system as demonstrated by intense staining with anti-actin, anti-tubulin, and anti-vimentin antibodies. When stained with anti-DEN-2 antibody, these cells showed a more diffuse pattern of fluorescence in the perinuclear and Golgi regions, consistent with ongoing virus replication. These large, strongly adherent cells were positive for nerve growth factor receptor, consistent with their identification as adventitial reticular cells. The molecule that mediates the virus interaction with susceptible cells has not previously been identified. Using plasma membrane proteins isolated from K562 cells, virus-binding studies suggest that an approximately 100-kD membrane protein may be involved in the initial virus-cell interaction.

Antigens, Viral↗
PubMed1997

Dengue hemorrhagic fever in Jakarta, Indonesia in 1988: isolation of dengue virus from patient whole blood using cell cultures.

During an outbreak of dengue hemorrhagic fever (DHF) in Jakarta, Indonesia in 1988, we attempted to isolate dengue virus using mosquito cells and a medium containing heparin. Whole blood, immediately after being drawn from patients, was inoculated into Aedes albopictus cell cultures temporarily maintained in the heparin-containing medium. The overall virus isolation rate was 25% (17 of 69) samples collected within three days after admission of the patients to hospital. No virus was obtained thereafter. The successful virus isolation was apparently not related to titers of anti-dengue virus hemagglutination-inhibiting antibodies present in patients' sera. The viruses were recovered from cases of each of the four World Health Organization grades of DHF without significant differences. The technique is simple and easily performed at bedside.

Aedes↗
PubMed1999

Type-3 dengue viruses responsible for the dengue epidemic in Malaysia during 1993-1994.

To characterize the dengue epidemic that recently occurred in Malaysia, we sequenced cDNAs from nine 1993-1994 dengue virus type-3 (DEN-3) isolates in Malaysia (DEN-3 was the most common type in Malaysia during this period). Nucleic acid sequences (720 nucleotides in length) from the nine isolates, encompassing the precursor of membrane protein (preM) and membrane (M) protein genes and part of the envelope (E) protein gene were aligned with various reference DEN-3 sequences to generate a neighbor-joining phylogenetic tree. According to the constructed tree, the nine Malaysian isolates were grouped into subtype II, which comprises Thai isolates from 1962 to 1987. Five earlier DEN-3 virus Malaysian isolates from 1974 to 1981 belonged to subtype I. The present data indicate that the recent dengue epidemic in Malaysia was due to the introduction of DEN-3 viruses previously endemic to Thailand.

Amino Acid Sequence↗
PubMed1998

An epidemic of dengue hemorrhagic fever and dengue shock syndrome in children in Delhi.

OBJECTIVE: To study clinical profiles and outcome of children of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) during 1996 Delhi epidemic. DESIGN: Retrospective study. SETTING: Hospital based study. METHODS: Children hospitalized from September to November 1996 were studied. All patients were diagnosed, managed and monitored according to a standard protocol. RESULTS: One hundred and thirty four children (80 (60%) males and 54 (40%) females) were studied. Sixty (45%) children were less than 6 years of age of which 12 presented during infancy. There were 92 (67%) cases of DHF and 42 (33%) cases of DSS. Common symptoms were fever (93%), abdominal pain (49%) and vomiting (68%). The commonest hemorrhagic manifestation was hematemesis (39%) followed by epistaxis (36%) and skin bleeds (33%). Hepatomegaly was observed in 97 (72%) cases and splenomegaly in 25 (19%). Serology was positive (IgM hemaglutination antibody titres > 1: 160) for dengue type 2 in 31 (80%) of 39 patients in whom sera was tested during the acute phase of illness. Mortality was 6%. Hematocrit > 40% was observed in only 25 (18%) patients and hence the management protocol was based on clinical signs and symptoms and not on hematocrit. CONCLUSIONS: A management protocol of DHF/DSS in which fluid therapy is not based on haematocrit values needs to be formulated.

Abdominal Pain↗
PubMed2000

[Dengue fever and dengue hemorrhagic fever].

Dengue virus is transmitted by mosquitoes and causes dengue fever/dengue haemorrhagic fever throughout the tropical areas of the world. There is an increasing incidence of dengue infections. Because of increasing travel activity, infection among Danes travelling abroad as well as imported cases are expected to be seen more frequently. In this review we describe the clinical manifestations, diagnosis, pathogenesis, treatment and prevention of the disease.

Animals↗
PubMed2004

Analysis of recombinant dengue virus polypeptides for dengue diagnosis and evaluation of the humoral immune response.

Dengue is a serious cause of morbidity and mortality in tropical areas worldwide. We cloned and expressed recombinant polypeptides spanning the entire genome of a Brazilian dengue virus type 2 (DENV-2) strain in contiguous segments to generate antigens for dengue diagnosis and evaluation of the human humoral immune response. When analyzed by Western blot and an enzyme-linked immunosorbent assay (ELISA) using human sera, the most reactive polypeptide (pD2-3(E)) was located in the N-terminal portion of the envelope protein. The sensitivity of an IgG-ELISA using pD2-3(E) versus mouse brain antigen was 100% with convalescent sera and 79% with acute sera, with a specificity of 100%. Sera from patients infected with other DENV serotypes recognized pD2-3(E) equally well, whereas sera positive for yellow fever, rubella, and measles showed little or no reactivity. Using this novel approach, we identified a candidate antigen to facilitate diagnosis of DENV infections and observed a surprising variability in antibody patterns in the clinical response to DENV infections.

Antibodies, Viral↗
PubMed2006

Dengue hemorrhagic Fever caused by sequential dengue 1-3 virus infections over a long time interval: Havana epidemic, 2001-2002.

A dengue epidemic caused by dengue virus 3 (DENV-3) occurred in Cuba in 2001-2002. It included cases of dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS). We report neutralizing antibody studies on sera from 54 of 78 DHF/DSS patients that provide evidence of infections occurring in the sequence DENV-1 followed by DENV-3. No sera showed infection in the sequence DENV-2 followed by DENV-3. Some sera showed a pattern of infection in the sequence DENV-1 followed by DENV-2 and then DENV-3. However definitive categorization of a tertiary infection was not possible because of broadly reactive antibodies, which could have been raised by infections in the sequence DENV-1 then DENV-3.

Antibodies, Viral↗
PubMed1982

Neutralization characteristics of dengue viruses isolated in mosquito cell culture and suckling mouse brain from a case of laboratory dengue infection.

One of the authors (Y.O.) of this paper was accidentally infected in the laboratory with dengue type 4 virus, isolated in Thailand in 1978 using the C6/36 clone of Singh's Aedes albopictus cells (SA). Two strains of the virus were isolated from Y.O.'s serum on the first day of illness using SA and suckling mouse brain (SMB). The SA-isolate and the infecting virus were neutralized with high efficiency by Y.O.'s convalescent sera, but the SMB-isolate was neutralized much less efficiently by the same sera. When the hosts of the isolates were exchanged (SA to SMB, and vice versa), the neutralization efficiencies were reversed. For analysis of this phenomenon, prototype dengue type 4 virus exclusively passed in SMB and the same type of virus passaged 10 times in SA were compared with the initial SA- and SMB-isolates in neutralization tests with sera from patients with dengue hemorrhagic fever (DHF) obtained in Thailand. The two strains of the prototype virus and the SA-isolate had similar high reactivities, but the SMB-isolate had low reactivity with sera of the DHF patients.

Aedes↗
PubMed1993

Dengue virus and endothelial cell: a related phenomenon to thrombocytopenia and granulocytopenia in dengue hemorrhagic fever.

Adhesion to endothelial cells by blood cells was assessed by measuring the cell number of each blood cell component in the supernatant after exposing blood cells to dengue-infected endothelial cells for 0, 10, 20 and 30 minutes. White blood cells, neutrophils, lymphocytes, platelets, and large lymphocytes or large unstained cells (LUC) preferentially bound to dengue-infected endothelial cells as compared to the control endothelial cells. P values were 0.0096 for total leukocytes and platelets, 0.006 for lymphocytes, and 0.001 for neutrophils and LUC. Monocytes basophils and eosinophils had no interaction with dengue-infected endothelial cells. The increased binding of neutrophil and platelet to endothelial cell may explain neutropenia and thrombocytopenia in DHF patients.

Agranulocytosis↗