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PubMed2006

A dominant role for FcgammaRII in antibody-enhanced dengue virus infection of human mast cells and associated CCL5 release.

Dengue virus is a major mosquito-borne human pathogen with four known serotypes. The presence of antidengue virus antibodies in the serum of individuals prior to dengue virus infection is believed to be an important risk factor for severe dengue virus disease as a result of the phenomenon of antibody-dependent enhancement operating on Fc receptor (FcR)-bearing cells. In addition to blood monocytes, mast cells are susceptible to antibody-enhanced dengue virus infection, producing a number of inflammatory mediators including IL-1, IL-6, and CCL5. Using the human mast cell-like lines KU812 and HMC-1 as well as primary cultures of human cord blood-derived mast cells (CBMC), we aimed to identify the participating FcRs in antibody-enhanced mast cell dengue virus infection, as FcRs represent a potential site for therapeutic intervention. CBMC expressed significant levels of FcgammaRI, FcgammaRII, and FcgammaRIII, and mast cell-like HMC-1 and KU812 cells expressed predominantly FcgammaRII. All four serotypes of dengue virus showed antibody-enhanced binding to KU812 cells. Specific FcgammaRII blockade with mAb IV.3 was found to significantly abrogate dengue virus binding to KU812 cells and CBMC in the presence of dengue-specific antibody. Dengue virus infection and the production of CCL5 by KU812 cells were also inhibited by FcgammaRII blockade.

Antibodies, Monoclonal↗
PubMed2006

[Reliability of the final dengue diagnosis in the epidemic occurring in Rio de Janeiro, Brazil, 2001-2002].

This study analyzed the reliability of the final diagnosis in the 155,242 dengue reports during the 2001-2002 epidemic in the city of Rio de Janeiro, Brazil, using the official information system on communicable diseases (SINAN). The system allows the following options for the final diagnosis: classic dengue, dengue hemorrhagic fever, discarded, inconclusive, and unknown. We built a classification routine in Epi Info to compare the final diagnosis from SINAN with Ministry of Health criteria. According to the final diagnosis, the case breakdown was: 52.4% classic dengue; 0.6% dengue hemorrhagic fever; 0.9% discarded; 46% inconclusive and unknown. The revised diagnosis showed that 78% of classic dengue, 69% of dengue hemorrhagic fever, and 21.1% of discarded cases met the classification criteria. Although the reliability of the SINAN final diagnosis was generally satisfactory (kappa = 0.681; 95%CI: 0.685-0.677), it was worse for fatal cases (kappa = 0.152; 95%CI: 0.046-0.258). Considering the epidemic's magnitude, the final diagnosis of classic dengue and dengue hemorrhagic fever was satisfactory, but the high proportion of inconclusive or unknown cases and the poor quality of information for fatal cases limit the usefulness of SINAN in this context.

Brazil↗
PubMed2000

[Development of immunizing agents against dengue].

The four serotypes of dengue flaviviruses are transmitted mainly by the Aedes aegypti mosquito, and some epidemics have been attributed to Ae. albopictus, Ae. polynesiensis, and various species of the Ae. scutellaris complex. The risk factors involved in dengue mortality and morbidity are related to the human host (genetic characteristics of infected persons; lifestyles, immune status, and health conditions of people; basic sanitation of dwellings; and water supply) and to the virus (genetic variability between and among serotypes, different pathogenicities, and geographic distribution). Notwithstanding the lack of knowledge of the immunopathobiology of dengue fever, important advances have been made in terms of a protective immune response, using attenuated dengue viruses or antigens produced by means of recombinant technologies. Efforts have been made since the 1940s to develop dengue vaccines. Immunity acquired from natural infection is specific for each serotype, and as many as three different serotype infections have been reported in one individual. For this reason, a tetravalent vaccine may likely be needed. Candidate vaccines against the four serotypes have been tested in volunteers and have proven to be immunogenic and safe. Although attenuated live virus vaccines are promising, more study is needed regarding their effectiveness and safety. Currently, several studies are ongoing to develop dengue vaccines using antigens from structural proteins (particularly E glycoprotein) and nonstructural proteins, with recombinant DNA technology and other biomolecular technologies. With the same goal, various expression vectors are being used, including Escherichia coli, baculovirus, vaccinia virus, and yellow fever virus. Unfortunately, no satisfactory results have been obtained in humans. The need for effective dengue vaccines is great, given the serious worldwide problem of the transmission of the four serotypes. Effective immunization against dengue would contribute to its prevention, with a positive cost-benefit relationship. Endemic dengue affects young children, and they should be immunized through the Expanded Program on Immunization.

Antigens, Viral↗
PubMed2006

Seroprevalence of dengue virus antibodies in asymptomatic Costa Rican children, 2002-2003: a pilot study.

OBJECTIVES: Since 1993 dengue has become more frequent in Costa Rica. Adults have been the most affected population, while children have remained virtually unharmed. So far no studies have investigated how many asymptomatic children have been affected by this virus. This pilot study documents the seroprevalence, measured as the presence of IgG antibodies, of dengue virus in asymptomatic children from two different geographical areas. METHODS: This descriptive, prospective epidemiologic study compared the presence of antibodies in children who live in a coastal region of a tropical country where dengue is endemic, and an inland area where dengue is not endemic. An enzyme-linked immunosorbent assay was used to test the serum for dengue virus IgG antibodies. None of the children had a prior history of dengue, fever, immunosuppressive therapy or underlying disease. RESULTS: During the period from July 2002 to July 2003, 103 children were recruited from each area. In the costal region we found a seroprevalence of 36.9%. In the inland area seroprevalence was 2.9%. CONCLUSIONS: We found a substantial number of asymptomatic infections in Costa Rican children. This greatly increases the risk of dengue hemorrhagic fever or dengue shock syndrome in these children, in whom previous dengue infection had gone undetected. Preventive efforts should be targeted at the costal region due to the higher prevalence in this area.

Antibodies, Viral↗
PubMed2006

Dengue virus induces expression of CXC chemokine ligand 10/IFN-gamma-inducible protein 10, which competitively inhibits viral binding to cell surface heparan sulfate.

Dengue virus is an arthropod-borne flavivirus that causes a mild febrile illness, dengue fever, or a potentially fatal syndrome, dengue hemorrhagic fever/dengue shock syndrome. Chemokines primarily orchestrate leukocyte recruitment to the areas of viral infection, which makes them critical mediators of immune and inflammatory responses. In the present study, we investigated the induction and function of chemokines in mice early after infection with dengue virus in vivo. We found that CXCL10/IFN-gamma-inducible protein 10 (IP-10) expression was rapidly and transiently induced in liver following infection. The expressed CXCL10/IP-10 likely mediates the recruitment of activated NK cells, given that anti-CXCL10/IP-10-treated mice showed diminished NK cell infiltration and reduced hepatic expression of effector molecules in activated NK cells after dengue virus infection. Of particular interest, we found that CXCL10/IP-10 also was able to inhibit viral binding to target cells in vitro. Further investigation revealed that various CXCL10/IP-10 mutants, in which the residues that mediate the interaction between the chemokine and heparan sulfate were substituted, failed to exert the inhibitory effect on dengue binding, which suggests that CXCL10/IP-10 competes with dengue virus for binding to heparan sulfate on the cell surface. Moreover, subsequent plaque assays showed that this inhibition of dengue binding blocked viral uptake and replication. The inhibitory effect of CXCL10/IP-10 on the binding of dengue virus to cells may represent a novel contribution of this chemokine to the host defense against viral infection.

Animals↗
PubMed1983

Original antigenic sin in dengue.

Sequential blood samples were obtained from eight Thai children before, during and 3-5 months after hospitalization for dengue shock syndrome. All patients experienced a secondary-type antibody response as evidenced by hemagglutination-inhibition antibody responses in acute and convalescent sera. Dengue 2 viruses were recovered from two patients. In their pre-illness blood sample, all children had monotypic neutralizing antibodies; five to dengue 1, two to dengue 3 and one to dengue 4. The highest neutralizing antibody titers in acute phase and late convalescent sera were to the initial infecting virus type. This report documents for sequential dengue infections the existence of an original antigenic sin antibody response. It may be possible to apply this phenomenon to identify initial dengue serotype infection in individuals experiencing secondary dengue infections, thus helping to clarify the antecedents to dengue shock syndrome.

Adolescent↗
PubMed1984

Mosquito cell cultures and specific monoclonal antibodies in surveillance for dengue viruses.

During the fall of 1981, a new method for the routine isolation and identification of dengue viruses in Puerto Rico was implemented utilizing C6/36 cell cultures and serotype specific antidengue monoclonal antibodies. A blind comparison of the monoclonal antibody indirect fluorescent antibody test (IFAT) and the complement fixation (CF) test for identification of 89 newly isolated dengue viruses of all four serotypes from the Caribbean, Asia and Africa showed 100% agreement. Although virus isolation rates were slightly lower than with the mosquito inoculation technique, use of the C6/36 cell culture system was much less time-consuming and allowed the processing of larger numbers of sera. Beginning in November 1981, a new virologic surveillance system was begun in Puerto Rico. Acute sera from persons with suspected dengue were selected for virus isolation attempts on the basis of geographic area of residence on the island, day after onset the blood was taken and clinical signs and symptoms. These sera were processed for virus isolation in C6/36 cell cultures, and virus isolates were identified by the IFAT using the monoclonal antibodies. Using this system, 2,702 sera were tested from November 1981 through August 1982. Dengue virus was isolated from 518, for an isolation rate of 19.2%. Dengue 1 was the predominant virus until December 1981, when dengue 4 became dominant. The changing patterns of dengue 1 and 4 distribution by time and geographic location on Puerto Rico were followed. This system allows the dengue viruses being transmitted in an area to be monitored with a minimal amount of effort and provides the early warning capability necessary to predict epidemic dengue.

Aedes↗
PubMed1996

Dengue encephalitis: a true entity?

Involvement of the central nervous system in dengue fever and dengue hemorrhagic fever has always been thought to be secondary to vasculitis with resultant fluid extravasation, cerebral edema, hypoperfusion, hyponatremia, liver failure, and/or renal failure. Thus, the condition has been referred to as dengue encephalopathy. Encephalitis or direct involvement of the brain by the virus was thought to be unlikely. This paper reports on six children who were seen over a period of two years presenting on the second or third day of illness with dengue encephalitis. The diagnosis was based upon a clinical picture of encephalitis and confirmed by cerebrospinal fluid (CSF) microscopy and electroencephalography changes. All six cases were confirmed dengue infections. Dengue 3 virus was isolated from the CSF of four cases and in one case, dengue 2 was detected by the polymerase chain reaction in both the CSF and blood. In the sixth case, virologic evidence was negative but dengue immunoglobulin M was detected in the CSF and blood. Since the onset of encephalitis appears early in the course of illness coinciding with the viremic phase, we postulate that the virus crosses the blood-brain barrier and directly invades the brain causing encephalitis. This study provides strong evidence that dengue 2 and 3 viruses have neurovirulent properties and behave similarly to other members of the Flaviviridae.

Antibodies, Viral↗
PubMed1999

Hospitalizations for suspected dengue in Puerto Rico, 1991-1995: estimation by capture-recapture methods. The Puerto Rico Association of Epidemiologists.

Capture-recapture estimations compare the results of 2 or more independent surveillance systems for the same event, and by measuring the degree of overlap between them, provide an estimate of the total number of events, and therefore the completeness of ascertainment in each system. The Puerto Rico Department of Health and the Dengue Branch of the Centers for Disease Control and Prevention (CDC) monitor dengue activity in Puerto Rico through 2 distinct surveillance systems: diagnostic specimens from patients with suspected dengue and infection control nurses' reports on patients hospitalized for suspected dengue. The patient listings from these systems were used in a 2-sample, capture-recapture calculation to estimate the total number of persons with suspected dengue hospitalized from 1991 to 1995. The laboratory positivity rate for suspected dengue cases who submitted appropriately timed serum samples in those years ranged from 72.1% to 81.2%. The laboratory-based (diagnostic sample) surveillance system (routinely used to monitor hospitalizations for suspected dengue) detected an average of 1,197 hospitalized cases during non-epidemic years, and 4,329 cases during the epidemic year of 1994. The detection rate of this system averaged 42% of the numbers derived by the capture-recapture method. In non-epidemic years, an estimated average of 2,791 patients (range = 1,553-3,481) was estimated to have been hospitalized with a clinical diagnosis of dengue, compared with 9,479 during 1994. These results demonstrate the under-detection inherent in passive surveillance systems for hospitalized cases of suspected dengue, and illustrate the value of capture-recapture techniques to better estimate the true incidence of hospitalizations for this disease.

Adult↗
PubMed1999

Application of molecular typing techniques in the 1998 dengue epidemic in Nicaragua.

This report presents the results of applying the reverse transcriptase-polymerase chain reaction (RT-PCR) to the analysis of clinical specimens during the 1998 dengue epidemic in Nicaragua. The RT-PCR was validated through comparison with viral isolation, resulting in a sensitivity of 100% and a specificity of 90%. In-country application of the RT-PCR permitted the rapid identification of dengue-3 virus as the cause of the epidemic at the beginning of 1998 and the detection of the reintroduction of dengue-2 virus in the middle of the year. Nineteen isolates of dengue-3 and one of dengue-2 were characterized using the restriction site-specific (RSS)-PCR technique. This showed that the dengue-3 strain belonged to the "Sri Lanka" subtype and that the dengue-2 strain belonged to the "Jamaica" subtype, both of which have been associated with hemorrhagic dengue in the Americas. The application of these simple PCR-based strain typing methods in a country endemic for dengue virus infections can help to characterize the transmission dynamics of this important emerging infectious disease problem and provide this information to local health authorities in a timely manner so that appropriate control measures can be implemented.

Dengue↗
PubMed2005

Rapid identification of dengue virus by reverse transcription-polymerase chain reaction using field-deployable instrumentation.

Dengue virus universal and dengue serotype 1 to 4, fluorogenic probe hydrolysis (TaqMan), reverse transcription-polymerase chain reaction assays were developed for screening and serotype identification of infected mosquito vectors and human sera using a field-deployable, fluorometric thermocycler. Dengue universal and dengue 1 to 4 serotype assay in vitro sensitivity and specificity results were 100% concordant when tested with total nucleic acid extracts of multiple strains of dengue serotype 1 to 4, yellow fever, Japanese encephalitis, West Nile, and St. Louis encephalitis viruses. The in vitro sensitivity and specificity results for all five assays were concordant when tested with a blind panel of 27 dengue virus-infected mosquitoes, 21 non-dengue (yellow fever, West Nile, or St. Louis encephalitis) flavivirus-infected mosquitoes, and 11 uninfected mosquitoes and with clinical specimens consisting of a human serum panel of eight dengue viremic and 31 non-dengue-infected febrile patient serum samples. No cross-reaction occurred with vector species or human genomic DNA. Sample processing and polymerase chain reaction required <2 hours.

Animals↗
PubMed2002

[Aedes aegypti, vector of the dengue virus: spatio-temporal structure of its genetic variation].

Aedes aegypti is the main vector of dengue viruses. Methods for limiting the spread of dengue outbreaks are currently based on vector control. Estimates of population genetic organization and gene flow with respect to vector capacity have provided great insights into dengue epidemiology. In Vietnam, dengue hemorrhagic fever was detected in the 1950's and becomes today the major problem of public health. Among factors influencing dengue epidemiology, ecological disturbances have a direct impact on vectorial system functioning. Human activities through urbanization creating sanitary conditions are convenient to the vector proliferation and then, to dengue endemisation. To assess the role of the vector in the changing pattern of dengue in South-East Asia, we studied the genetic differentiation and the vector competence towards dengue 2 virus at two scales: at a spatial level (a local scale (i.e., Ho Chi Minh City) and a regional scale (i.e., Cambodia, Thailand and South Vietnam) and at a temporal scale. This study allows to propose a model of Ae. aegypti population functioning according space and time. Dynamics of dengue virus diffusion in relation with the vector, depend on the population genetic composition and its evolution.

Aedes↗
PubMed1992

Immunity and immunopathology in dengue virus infections.

Dengue virus infections are a serious public health problem in tropical and subtropical areas of the world. Based on epidemiological data, it has been postulated that immune responses to dengue virus contribute to the pathogenesis of severe dengue illness, dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Host immune responses are also important for controlling dengue virus infection. Therefore, dengue virus infections are an interesting model to explore the interactions between viruses and the immune system which result in immunopathology or recovery from infection. In this paper, we review immune responses to dengue viruses with an emphasis on the human T cell responses, and discuss possible roles of these immune responses in the control of dengue virus infection and in the pathogenesis of DHF/DSS.

Adolescent↗
PubMed2006

Molecular characterization and clinical evaluation of dengue outbreak in 2002 in Bangladesh.

During the febrile illness epidemic in Bangladesh in 2002, 58 people died out of the 6,132 affected. Two hundred hospitalized patients were analyzed clinically, serologically and virologically to determine the features of this dengue infection. Among the 10- to 70-year-old age group of the 200 clinically suspected dengue patients, 100 (50%) were confirmed as dengue cases by virus isolation and dengue IgM-capture ELISA. Of the 100 dengue-confirmed cases, the mean age was 29.0 (+/-12.4). The possible dengue secondary infection rate determined by Flavivirus IgG-indirect ELISA was 78% in 2002. Eight dengue virus strains were isolated, representing the first dengue virus isolation in the country, and all of the strains were dengue virus type-3 (DEN-3). Sequence data for the envelope gene of the DEN-3 Bangladeshi isolates were used in a phylogenetic comparison with DEN-3 from other countries. A phylogenetic analysis revealed that all 8 strains of DEN-3 were clustered within a well-supported independent sub-cluster of genotype II and were closely related to the Thai isolates from the 1990s. Therefore, it is likely that the currently circulating DEN-3 viruses entered Bangladesh from neighboring countries.

Adolescent↗
PubMed2006

[The clinical behavior of dengue in patients hospitalized in the Antonio Roldán Betancur Hospital of Apartadó, Antioquia, 2000].

INTRODUCTION: Dengue has exhibited an epidemic behaviour in Apartad6 since 1998. OBJECTIVE: To describe the clinical behavior of dengue in patients hospitalized with this diagnosis in the Antonio Roldán Betancur Hospital of Apartadó in the year 2000. MATERIALS AND METHODS: Descriptive retrospective study with variables obtained from clinical records of patients hospitalized with dengue in the Antonio Roldán Betancur Hospital of Apartadó. RESULTS: 45 patients with the diagnosis of dengue were hospitalized during the study period, all of them with IgM antibodies against dengue virus. Three (6.7%) patients were classified as classic dengue fever, 22 (48.9%) as dengue fever with haemorrhagic manifestations, and 20 patients (44.4%) were classified as dengue haemorrhagic fever. Patient age ranged from 4 months to 68 years, with a mean of 22.1 years. The great majority of patients (82.8%) came from urban areas. The most frequent clinical manifestations were fever (100%), myalgia (71.1%), joint pain (71.1%), headache (70%) and abdominal pain (67.5%). The most frequent haemorrhagic manifestations were petechiae (60%), hematuria (31.7%), ecchymoses on venipuncture sites (24.4%), gastric bleeding (22.2%) and abnormal vaginal bleeding (15.5%). Platelet count ranged from 11.000 to 186.000/mm3, with a mean of 48.688. Hematocrit ranged from 26% to 59% with a mean of 41.5%. Four of the 45 patients died. 40% of patients had other co-morbidities; coinfection with malaria was the most frequent amongst them (8 patients, 17.7%). CONCLUSION: Two remarkable findings of our descriptive retrospective study were the high mortality of dengue in our population (8.8%) 4/45, specially in children under 1 year of age (3/4) and the high incidence of simultaneous infection with malaria parasites.

Adolescent↗
PubMed1995

Dengue serologic survey in Ribeirão Preto, São Paulo, Brazil.

The city of Ribeirão Preto suffered a dengue 1 epidemic that began in November 1990 and ended in March 1991. A serologic survey designed to detect IgG antibodies to the four dengue serotypes and other flaviviruses was carried out in Ribeirão Preto during September and October of 1992. Dengue 1 antibodies were detected in 5.4% of the survey participants. Significantly higher seropositivity (9.3%) was found among subjects residing in the Northwest Sector of Ribeirão Preto than among those living in the city's other three sectors. The Northwest Sector also exhibited relatively high levels of breeding sites used by the Aedes aegypti vector, the highest number of reported dengue cases of any sector, and relatively poor socioeconomic conditions. The fact that the epidemic was limited mainly to the Northwest Sector probably resulted mainly from intense vector control and educational measures undertaken in response to the outbreak. As of the 1992 survey, most of the city's population remained vulnerable to dengue 1 infection; however, an estimated 23,000 with dengue 1 antibodies appeared to be at relatively high risk of developing dengue hemorrhagic fever/dengue shock syndrome in the event of infection with dengue 2. Both of these considerations indicate an ongoing need to maintain dengue education and vector control measures.

Adolescent↗
PubMed1994

Storage stability of dengue IgM and IgG antibodies in whole blood and serum dried on filter paper strips detected by ELISA.

Blood specimens from 133 patients clinically diagnosed as dengue virus infection by physicians in Nakhon Phanom Hospital, Thailand, were examined to detect anti-dengue IgM and IgG antibodies by antibody capture ELISA. The blood specimens were divided into 3 types of storage; (1) frozen serum aliquots, (2) whole blood dried on filter paper strips, and (3) sera dried on filter paper strips. These specimens were stored for the periods of 1, 3, 4, and 5 months, at -20 degrees C in the case of frozen serum aliquots, or at room temperature in the case of specimens dried on filter paper strips, before examined in paralleled by the ELISA. Anti-dengue IgG antibodies were stable for at least 5 months of storage as dried whole blood or serum on filter paper strips. So were the anti-dengue IgM antibodies in the dried whole blood from secondary dengue cases. Anti-dengue IgM antibodies from primary dengue cases declined slowly in whole blood and more rapidly in serum, both dried on filter paper strips. In the serum dried on filter paper strips, even anti-dengue IgM antibodies from secondary cases decreased significantly on storage. We suggest that diagnosis on dengue infections by IgM-capture ELISA should be performed within 1 month after the test specimens are collected as whole blood, not as serum, when the filter paper method is used for sample collection.

Antibodies, Viral↗
PubMed1994

The first epidemic of dengue hemorrhagic fever in French Guiana.

From July 1991 to October 1992, an outbreak of dengue spread into the main urban areas of French Guiana, where 90% of the country's 114,808 inhabitants live. In mid-July 1991 dengue-2 virus was identified as being responsible for most cases, while dengue-1 virus was rarely isolated and circulated at a low level. The number of dengue cases during this period was unknown because there was no clinically based dengue surveillance system. The only available data were for the number of suspected cases as indicated by the number of patients for whom blood samples were submitted to a laboratory for dengue diagnosis. Eight hundred forty-seven of the 2,948 suspected cases were diagnosed in the laboratory as dengue cases. Six fatal cases were reported. This outbreak was marked by the appearance of the first clinical cases of dengue hemorrhagic fever (DHF) in French Guiana. Forty cases met the World Health Organization definition of clinical DHF: 32 were grade II, seven were grade III, and one was grade IV and fatal. Eighteen cases were confirmed in the laboratory and 12 were probable; there was no proof of the dengue etiology for the remaining patients.

Antibodies, Viral↗