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PubMed1999

Bystander target cell lysis and cytokine production by dengue virus-specific human CD4(+) cytotoxic T-lymphocyte clones.

Dengue hemorrhagic fever, the severe form of dengue virus infection, is believed to be an immunopathological response to a secondary infection with a heterologous serotype of dengue virus. Dengue virus capsid protein-specific CD4(+) cytotoxic T-lymphocyte (CTL) clones were shown to be capable of mediating bystander lysis of non-antigen-presenting target cells. After activation by anti-CD3 or in the presence of unlabeled antigen-presenting target cells, these clones could lyse both Jurkat cells and HepG2 cells as bystander targets. Lysis of HepG2 cells suggests a potential role for CD4(+) CTL in the liver involvement observed during dengue virus infection. Three CD4(+) CTL clones were demonstrated to lyse cognate, antigen-presenting target cells by a mechanism that primarily involves perforin, while bystander lysis occurred through Fas/Fas ligand interactions. In contrast, one clone used a Fas/Fas ligand mechanism to lyse both cognate and bystander targets. Cytokine production by the CTL clones was also examined. In response to stimulation with D2 antigen, CD4(+) T-cell clones produced gamma interferon, tumor necrosis factor alpha (TNF-alpha) and TNF-beta. The data suggest that CD4(+) CTL clones may contribute to the immunopathology observed upon secondary dengue virus infections through direct cytolysis and/or cytokine production.

Antigen Presentation↗

PubMed2002

Increased production of interleukin-8 in primary human monocytes and in human epithelial and endothelial cell lines after dengue virus challenge.

The more severe form of dengue virus infection, dengue hemorrhagic fever, is characterized by plasma leakage and derangements in hemostasis. As elevated interleukin-8 (IL-8) levels have been observed in sera from patients with more severe disease manifestations, a study was initiated to look at the effect of dengue virus infection in vitro on proinflammatory cytokine secretion and expression. A significant increase in IL-8 levels in the culture supernatant of primary human monocytes infected with dengue 2 virus (D2V) New Guinea C (NGC) was found by enzyme-linked immunosorbent assay. Additionally, by reverse transcriptase PCR, the mRNA was also augmented. Among the proinflammatory cytokines and their mRNAs measured (IL-6, IL-1 beta, IL-8, and tumor necrosis factor alpha), IL-8 showed the greatest change following D2V infection. Similarly, two cell lines, 293T (a human epithelial cell line) and ECV304 (an endothelial cell line), were permissive to D2V NGC and responded to the infection by increasing the synthesis of IL-8. Nuclear factor kappa B (NF-kappa B) and nuclear factor IL-6 (NFIL-6) are primary mediators of IL-8 expression. We studied the transcriptional regulation of IL-8 in the ECV304 and 293T cell lines and found that the induction of IL-8 gene expression involved the activation of NF-kappa B (P = 0.001) and, to a lesser extent, the activation of NFIL-6 in ECV304 cells only. We next observed by the chromatin immunoprecipitation procedure in vivo acetylation of core histones bound to the IL-8 promoter after D2V infection. IL-8 produced by infected monocytes and also IL-8 that may be produced by endothelial or other epithelial cells is associated with the hyperacetylation of histones bound to the IL-8 promoter in addition to the activation of transcription by NF-kappa B. We hypothesize that the overall increase in IL-8 synthesis observed in this in vitro study may play a role in the pathogenesis of the plasma leakage seen in dengue hemorrhagic fever and dengue shock syndrome.

Animals↗

PubMed2005

Heat shock protein 90 and heat shock protein 70 are components of dengue virus receptor complex in human cells.

Dengue virus requires the presence of an unidentified cellular receptor on the surface of the host cell. By using a recently published affinity chromatography approach, an 84-kDa molecule, identified as heat shock protein 90 (HSP90) by matrix-assisted laser desorption ionization-time of flight mass spectrometry, was isolated from neuroblastoma and U937 cells. Based on the ability of HSP90 (84 kDa) to interact with HSP70 (74 kDa) on the surface of monocytes during lipopolysaccharide (LPS) signaling and evidence that LPS inhibits dengue virus infection, the presence of HSP70 was demonstrated in affinity chromatography eluates and by pull-down experiments. Infection inhibition assays support the conclusion that HSP90 and HSP70 participate in dengue virus entry as a receptor complex in human cell lines as well as in monocytes/macrophages. Additionally, our results indicate that both HSPs are associated with membrane microdomains (lipid rafts) in response to dengue virus infection. Moreover, methyl-beta-cyclodextrin, a raft-disrupting drug, inhibits dengue virus infection, supporting the idea that cholesterol-rich membrane fractions are important in dengue virus entry.

Cell Line, Tumor↗

PubMed2006

[Dengue infection in children and adolescents in Panama].

BACKGROUND: Dengue is a serious emerging infectious disease and constitutes a major international health concern. MATERIAL AND METHODS: All reports of confirmed dengue infection in patients aged less than 18 years old between 2000 and 2005 were included. A confirmed diagnosis was established by culture of the virus within the first 3 days of symptom onset or by serologic assays 5-30 days after symptom onset. Clinical and epidemiological features were analyzed. RESULTS: A total of 457 patients were included (57.6 % female). The median age was 13 years (IQR 5 6). A greater number of cases were detected in urban areas and during the rainy season (May-November). Two epidemics were reported in 2001 (33.9 %) and the first eight months of 2005 (23.1 %). The most prevalent symptoms were fever (95.2 %), severe headache (74.2 %), chills (65.9 %), rash (63.5 %), myalgias (51.9 %) and retro-orbital pain (51.6 %). No significant differences were found between male and female patients. Significant differences in clinical features were found when the patients were divided into 3 groups; < 5 years old, 6-10 years old and > 10 years old. Fifty-three percent of the patients had had previous contact with a dengue-infected individual. There were 7 patients with dengue hemorrhagic fever, 4 of whom died. CONCLUSIONS: Dengue virus infection is still a major health problem in Panama. To achieve effective control of dengue, further epidemiological studies, such as our own, are needed to design appropriate preventive measures.

Adolescent↗

PubMed2005

Role of ultrasound in dengue fever.

This study was performed to find out whether ultrasound is an important adjunct to clinical and laboratory profile in diagnosing dengue fever or dengue haemorrhagic fever and to further determine whether ultrasound is useful in predicting the severity of the disease. Ultrasound was performed on 128 patients (2-9 years) with clinical suspicion of dengue fever. Serological tests were performed to confirm the diagnosis. 40 patients were serologically negative for dengue fever and later excluded from the study. Of the remaining 88 serologically positive cases, 32 patients underwent ultrasound on second to third day, repeated on fifth to seventh day of fever and in 56 patients ultrasound was done only on fifth to seventh day of fever. Of the 32 patients who underwent the study on second to third day of fever, all showed gall bladder wall thickening and pericholecystic fluid, 21% had hepatomegaly, 6.25% had splenomegaly and right minimal pleural effusion. Follow-up ultrasound on fifth to seventh day revealed ascites in 53% left pleural effusion in 22% and pericardial effusion in 28%. Of the 56 patients who underwent the study on fifth to seventh day of fever for the first time all had gall bladder wall thickening, 21% had hepatomegaly, 7% had splenomegaly, 96% had ascites, 87.5% had right pleural effusion, 66% had left pleural effusion and 28.5% had pericardial fluid. To conclude, in an epidemic of dengue, ultrasound features of thickened gall bladder wall, pleural effusion and ascites should strongly favour the diagnosis of dengue fever.

Ascitic Fluid↗

PubMed1998

Dengue fever epidemic potential as projected by general circulation models of global climate change.

Climate factors influence the transmission of dengue fever, the world's most widespread vector-borne virus. We examined the potential added risk posed by global climate change on dengue transmission using computer-based simulation analysis to link temperature output from three climate general circulation models (GCMs) to a dengue vectorial capacity equation. Our outcome measure, epidemic potential, is the reciprocal of the critical mosquito density threshold of the vectorial capacity equation. An increase in epidemic potential indicates that a smaller number of mosquitoes can maintain a state of endemicity of disease where dengue virus is introduced. Baseline climate data for comparison are from 1931 to 1980. Among the three GCMs, the average projected temperature elevation was 1.16 degrees C, expected by the year 2050. All three GCMs projected a temperature-related increase in potential seasonal transmission in five selected cities, as well as an increase in global epidemic potential, with the largest area change occurring in temperate regions. For regions already at risk, the aggregate epidemic potential across the three scenarios rose on average between 31 and 47% (range, 24-74%). If climate change occurs, as many climatologists believe, this will increase the epidemic potential of dengue-carrying mosquitoes, given viral introduction and susceptible human populations. Our risk assessment suggests that increased incidence may first occur in regions bordering endemic zones in latitude or altitude. Endemic locations may be at higher risk from hemorrhagic dengue if transmission intensity increases.

Aedes↗

PubMed2005

Nonstationary influence of El Niño on the synchronous dengue epidemics in Thailand.

BACKGROUND: Several factors, including environmental and climatic factors, influence the transmission of vector-borne diseases. Nevertheless, the identification and relative importance of climatic factors for vector-borne diseases remain controversial. Dengue is the world's most important viral vector-borne disease, and the controversy about climatic effects also applies in this case. Here we address the role of climate variability in shaping the interannual pattern of dengue epidemics. METHODS AND FINDINGS: We have analysed monthly data for Thailand from 1983 to 1997 using wavelet approaches that can describe nonstationary phenomena and that also allow the quantification of nonstationary associations between time series. We report a strong association between monthly dengue incidence in Thailand and the dynamics of El Niño for the 2-3-y periodic mode. This association is nonstationary, seen only from 1986 to 1992, and appears to have a major influence on the synchrony of dengue epidemics in Thailand. CONCLUSION: The underlying mechanism for the synchronisation of dengue epidemics may resemble that of a pacemaker, in which intrinsic disease dynamics interact with climate variations driven by El Niño to propagate travelling waves of infection. When association with El Niño is strong in the 2-3-y periodic mode, one observes high synchrony of dengue epidemics over Thailand. When this association is absent, the seasonal dynamics become dominant and the synchrony initiated in Bangkok collapses.

Climate↗

PubMedDate not supplied

Involvement of the central nervous system in dengue fever: three serologically confirmed cases from Fortaleza Ceará, Brazil.

Three cases of dengue fever involving the central nervous system (CNS) are reported. All occurred in 1994 during a dengue (DEN) epidemic caused by serotypes DEN-1 and DEN-2. The first case examined was a 17-year-old girl who complained of fever, nuchal rigidity and genital bleeding. Three blood samples were positive by anti-dengue IgM ELISA and showed hemagglutination-inhibition (HI) test titers > or = 1,280. The second case concerned a 86-year-old women with fever, muscle and joint pains, altered consciousness, syncope, nuchal rigidity and meningismus. Her blood sample showed an HI titer of 1:320 for flaviviruses, and an IgM ELISA positive for dengue. The third case was a 67-year-old women with fever, abnormal behaviour, seizures, tremor of extremities, thrombocytopenia, increased hematocrit and leukopenia. The patient suffered a typical case of dengue hemorrhagic fever with ensuing shock and a fatal outcome. A single blood sample showed HI antibodies of > or = 1,280 and an IgM ELISA positive for dengue. No virus could be isolated from any patient by inoculation of blood into C6/36 cells and suckling mice. No other agent of disease was encountered in the patient.

Adolescent↗

PubMedDate not supplied

[Dengue: a new approach].

Previous definition of classic dengue, with or without bleeding, and of dengue hemorrhagic fever (DHF) that may evolve without bleeding and with or without dengue shock syndrome (DSS) are reviewed here. The classical approach to the diagnosis and treatment of dengue, although useful in the past, nowadays breeds confusion and adds a burden to the physician's task of decision-making regarding the treatment of patients with severe forms of the disease. The classification of dengue proposed in this paper, and summarized in a diagram, incorporates new concepts about sepsis, systemic inflammatory response syndrome (SIRS), and acute respiratory distress syndrome (ARDS). This new approach, in our view, is a useful guide to initial evaluation and treatment of the disease. It also approximates the dengue syndrome to other protocols and medical procedures routinely used in intensive care units, making it easier to be followed by the health personnel working in areas subject to epidemic bursts.

Dengue↗

PubMedDate not supplied

Retrospective study on dengue in Fortaleza, state of Ceará, Brazil.

A retrospective serologic study was carried out in Fortaleza, State of Ceará, Brazil, in order to detect the dengue virus activity before recognizing the epidemic of 1994. Mac-Elisa was performed by using a mixture of specific DEN-1 and DEN-2 antigens on serum samples from the Emilio Ribas Laboratory collection. Samples were obtained from 1,224 patients with exanthematic febrile disease and negative serological results for rubella. All specimens were taken during November 1993 to May 1994. The results confirmed dengue infections in Fortaleza by November 1993, approximately six months before the beginning of the epidemic, proving how misleading diagnosis of dengue infection are still troublesome, in spite of the strong dengue activity in Ceará. The authors stress the urgent necessity to implement the active surveillance system in order to prevent another extensive dengue fever epidemics in the state. Epidemiological background of the dengue activity in the State of Ceará is described.

Brazil↗

PubMed2006

Dengue virus RNA polymerase NS5: a potential therapeutic target?

Dengue fever (DF)/dengue haemorrhagic fever (DHF) is the most common arthropod-borne viral infection, where it is now estimated that 2.5-3 billion people world-wide are at risk of infection. Currently there is no available treatment, in the form of vaccine or drug, making eradication of the mosquito vector the only viable control measure, which has proved costly and of limited success. There are a number of different vaccines undergoing testing, but whilst a dengue vaccine is clearly desirable, there are several issues which make live-attenuated vaccines problematic. These include the phenomenon of antibody-dependent enhancement (ADE) and the possibility of recombination of attenuated vaccine strains with wild-type flavivirus members reverting vaccines to a virulent form. Until we gain a better understanding of these issues and their associated risks, the safety of any live dengue vaccine cannot be assured. It therefore may be safer and more feasible for therapeutic-based approaches to be developed as an alternative to live vaccines. As our understanding of dengue molecular biology expands, new potential targets for drugs are emerging. One of the most promising is the dengue non-structural protein 5 (NS5), the largest and most highly conserved of the dengue proteins. This review examines the unique properties of NS5, including its functions, interactions, subcellular localisation and regulation, and looks at ways in which some of these may be exploited in our quest for effective drugs.

Active Transport, Cell Nucleus↗

PubMed2006

Etiology of exanthema in children in a dengue endemic area.

OBJECTIVE: To study the etiology of exanthema cases, with or without fever, in children seen in the emergency room of a hospital located in a region where dengue is endemic. METHODS: Enrollment took place between 21/09/2001 and 20/09/2002 and included 95.9% (71/74) of children presenting with exanthema at the emergency room of the Hospital Universitário de Campo Grande, MS (4.1% refusals). After the children had had their details taken and entered on the study protocol, they were subjected to physical examination followed by collection of blood samples for blood testing with platelet counts and serology (IgM and IgG); initially for dengue, rubella and toxoplasmosis and then, in negative cases, serology was also run for parvovirus, herpes virus type 6 and measles. RESULTS: Laboratory diagnoses were confirmed by means of IgM antibody assay in 88.7% of the cases investigated: dengue (77.5%), herpes virus type 6 (8.4%), parvovirus (2.8%) and in eight patients diagnosis was inconclusive (11.3%). On this occasion no positive serology (IgM) was observed for measles, rubella or toxoplasmosis. The most common clinical manifestations among the dengue patients were: fever, itching, prostration, myalgia and positive tourniquet test results. In 58.4% (32/55) of those cases diagnosed with dengue, the tourniquet test was positive, which was a statistically significant difference when compared with the remainder of the sample. CONCLUSIONS: When children present with exanthema, it is possible that dengue is the primary causative disease, depending on the epidemiology of the location. Constant control of epidemiological and serological surveillance of exanthematous diseases is necessary.

Adolescent↗

PubMed1976

Dengue virus replication enhancement in peripheral blood leukocytes from immune human beings.

Dengue 2 virus replication in peripheral blood leukocyte cultures from 10 of 13 Asian or Polynesian subjects with actively acquired dengue immunity and two of three infants with passively acquired dengue antibody. Only 2 of 11 cultures from nonimmune infants or children supported viral replication. This study establishes a parallel in the biological behavior of human and simian PBL with respect to the immunological dependence of dengue virus replication in vitro. Elucidation of the mechanism (or mechamisms) regulating growth of dengue virus in leukocytes from immune hosts may contribute to an understanding of the role that virus-leukocyte interactions play in the pathogenesis of human dengue illness.

Adolescent↗

PubMedDate not supplied

Dengue surveillance in Florida, 1997-98.

Recent dengue outbreaks in the Caribbean and Central and South America and the presence of competent mosquito vectors increase the likelihood of future autochthonous transmission in Florida. During April 1997 to March 1998, a laboratory-based active surveillance program detected 18 cases of dengue involving all four dengue serotypes. All patients reported recent travel to countries with indigenous dengue transmission. These results demonstrate that dengue infections are imported into Florida at a much higher rate than reflected by previous passive surveillance; therefore, the risk for local dengue transmission may be increasing.

Adolescent↗

PubMed1982

Identification of distinct antigenic determinants on dengue-2 virus using monoclonal antibodies.

Monoclonal antibodies directed against antigenic determinants of the New Guinea C strain of dengue-2 virus were obtained from lymphocyte hybridomas produced by fusing immune mouse lymphocytes with mouse myeloma cells. Hybridoma cell culture supernatants were screened by using a radioimmunoassay employing detergent-solubilized dengue-2 infected cell antigens. Monoclonal antibodies in ascitic fluids induced by 22 selected hybridomas were characterized by the hemagglutination-inhibition, plaque reduction neutralization, immunofluorescence, and complement-fixation tests. Both type-specific and broadly cross-reactive antibodies were observed, and immunoglobulin subclasses IgG1 and IgG2a were represented in both groups. At least three distinct antigenic determinants on the virion were defined using these antibodies. A single hybridoma produced antibody which recognized a dengue-2 virus type-specific determinant and exhibited high titered neutralization but had a low titer by hemagglutination inhibition. Four preparations reacted with a type-specific determinant and exhibited hemagglutination inhibition but did not neutralize. Seventeen hybridomas produced antibodies which were broadly cross reactive in all tests. Only two preparations reacted by complement fixation with dengue-2 antigens; both were cross reactive. Immunofluorescence specificity or cross reactivity correlated with neutralization and/or hemagglutination-inhibition. The dengue-2 virus type-specific antibody useful for identification of dengue-2 infected cells by immunofluorescence has been deposited in the Hybridoma Cell Bank of the American Type Culture Collection.

Animals↗

PubMed1985

Comparative susceptibility of mosquito species and strains to oral and parenteral infection with dengue and Japanese encephalitis viruses.

Thirty-four strains of Asian and Pacific mosquitoes belonging to 22 species of 7 genera were compared for oral and/or parenteral susceptibility to infection with 1 or more strains of each of the 4 dengue serotypes. Surprisingly, several species of common man-biting Aedes were much more susceptible to oral infection with each of the 4 dengue serotypes than was Aedes aegypti. These species included Aedes albopictus and members of the scutellaris group of the subgenus Stegomyia found on South Pacific islands. Mosquito strains and species relatively susceptible to 1 dengue serotype usually were relatively susceptible to the others also. Almost all species of Aedes tested were uniformly susceptible to parenteral infection with the dengue viruses but, with the exception of a species of Tripteroides, species of all other genera were comparatively resistant to that mode of infection. Dengue viruses usually replicated to about the same extent in orally-infected mosquitoes as they did in parenterally-infected specimens of the same species. Seventeen species of mosquitoes of 7 genera also were tested for parenteral susceptibility to infection with Japanese encephalitis virus. With the possible exception of 2 species of Anopheles, the virus replicated to about the same degree in all species tested and achieved levels considerably higher than did any of the dengue viruses in the same mosquito strain and species held under the same conditions.

Aedes↗

PubMed1997

Rapid characterization of genetic diversity among twelve dengue-2 virus isolates by single-strand conformation polymorphism analysis.

Single-strand conformation polymorphism (SSCP) analysis was used to characterize genetic polymorphisms among 12 isolates of dengue-2 virus, which were previously genetically characterized by RNase T1 oligonucleotide mapping and by sequencing the viral envelope (E) gene. Specific cDNA fragments from the dengue-2 isolates were amplified by the reverse transcriptase-polymerase chain reaction. The viral E, premembrane (prM), and nonstructural 5 (NS5) gene cDNAs of 291 basepairs (bp), 291 bp, and 201 bp, respectively, were denatured, rapidly chilled to promote intrastrand reassociation, electrophoretically separated on nondenaturing polyacrylamide gels, and SSCP patterns were observed by silver staining. The SSCP analysis revealed polymorphisms among a number of dengue-2 isolates from the same topotype, and these were markedly different between isolates of different topotype (distinct genetic group). Comparison of nucleotide sequence and SSCP analyses of the 291-bp E cDNA demonstrated that virus isolates that produced identical SSCP patterns contained 0-7 nucleotide substitutions, whereas isolates that showed different SSCP patterns contained 4-25 nucleotide substitutions. Positive predictive value and negative predictive value as measures of certainty for predicting identical and different sequences were 26% and 100%, respectively. The SSCP patterns of the 12 dengue-2 isolates suggested greater genetic variation in the prM gene region than in either the E or NS5 gene regions. The SSCP analyses should allow easy, sensitive, and rapid screening of dengue viruses isolates and the assessment of variation at a number of sites in the virus genome. Additionally, SSCP screening of dengue-2 virus for genetic variability may reveal the introduction of new viral genotypes in a given geographic area. These genetic variants of the virus could serve as markers of the epidemic potential of the virus strain.

DNA, Complementary↗

PubMed1998

The early use of break-bone fever (Quebranta huesos, 1771) and dengue (1801) in Spanish.

The appellation of break-bone fever for dengue is attributed to the popular name given to the disease in the first described epidemic of an illness that is clinically compatible with dengue in Philadelphia in 1780. The origin of the word dengue as the name of an illness is currently attributed to the Swahili phrase Ka dinga pepo, thought to have crossed from Africa to the Caribbean in 1827; in Cuba this phrase was popularly identified with the Spanish word dengue. This article presents documents from Spanish archives that indicate the use of quebranta huesos (break-bone) by a physician in Puerto Rico to describe a febrile illness in 1771, and the use of the term dengue by the Queen of Spain in 1801 to describe an acute febrile illness with bone and joint pains, hemorrhage, and jaundice. These texts are evidence of a more generalized use of the term break-bone than previously recognized, and conversely, the specifically Spanish origin of dengue as the name for an illness.

Dengue↗