Dengue Virus Antigen Research Articles

Immuno-proteomic interrogation of antigen presentation during Dengue and Zika infections reveals novel targets for the design of broad acting T-cell vaccines

Abstract The recent outbreak of Zika virus (ZIKV) infection across the Americas has raised dire concerns about its potential long-term health effects including its negative impact on neuronal, psychological and motor development in affected individuals. Emerging evidence pointing to Alzheimer’s-like damage to the adult brain further highlights the urgent need for efficacious prophylactic measures. Zika-affected regions are often endemic to the related Dengue virus (DENV). Antibodies against the two viruses are crossreactive and cause antibody-mediated enhancement (ADE) of infection. This confounds the design of traditional vaccines that elicit humoral antibody-mediated responses. T cell vaccines mediated by cytotoxic CD8+ T cells that recognize conserved viral epitopes presented on the major histocompatibility complex (MHC-I) of infected cells are unencumbered by ADE and can supplement or substitute traditional vaccines. In order to identify robust T cell vaccine candidates, we first used an immunoproteomic approach and isolated over 100 novel DENV and ZIKA epitopes presented by infected B lymphocytes in vitro. These viral peptides were not predicted to be epitopes by computational programs typically used for epitope discovery, which discard >95% of all predictions. Focussing on epitopes derived from polyprotein regions strictly conserved between the two viruses, we validated the presentation of these epitopes in primary dendritic cells and assayed their ability to stimulate robust T cell responses in ex vivo ELISpot assays. Together these discoveries let us assay the immunogenic potentials of an unprecedented range of DENV and ZIKV antigens which have the potential to serve as broad acting and effective Zika vaccine candidates.

Acute phase human B cell response after ZIKV infection

Abstract Prior to its emergence in Brazil in 2015, Zika virus (ZIKV) was mainly reported to cause either asymptomatic infections or mild, self-limiting illness. However, the disease pathology of recent epidemics includes neurological complications such as Guillain-Barré Syndrome in adults and congenital microcephaly in neonates. Consequently, there is a pressing need to understand ZIKV pathogenesis and the immunology of ZIKV infections. To this end, we have studied the B cell response in acutely-infected ZIKV patients, both at the serum and at the monoclonal level. Of the several plasmablast-derived mAbs generated, most were specific to the ZIKV envelope. Given the immunological cross-reactivity between ZIKV and dengue virus (DENV), we tested the panel for reactivity against DENV1-4 and found that all ZIKV-reactive mAbs also bound DENV antigens. Interestingly, even though the patients was infected with ZIKV, more mAbs neutralized DENV than ZIKV in vitro. Patient serum showed high ZIKV and DENV IgG binding titers, illustrating that this cross-reactivity was evident at the serum level as well. At the cellular level, the plasmablasts isolated from these patients exhibited high levels of somatic hypermutation, comparable to frequencies seen in secondary dengue cases. Together, these data suggest that the patients may have previously been exposed to DENV, and that the poorly neutralizing mAbs generated after ZIKV infection could be the product of a DENV-derived memory response, suggesting evidence of original antigenic sin (OAS). These findings have important implications from a public health viewpoint, given that a majority of areas currently experiencing ZIKV outbreaks are also DENV endemic.

Pathogen-specific deep sequence-coupled biopanning: A method for surveying human antibody responses.

Identifying the targets of antibody responses during infection is important for designing vaccines, developing diagnostic and prognostic tools, and understanding pathogenesis. We developed a novel deep sequence-coupled biopanning approach capable of identifying the protein epitopes of antibodies present in human polyclonal serum. Here, we report the adaptation of this approach for the identification of pathogen-specific epitopes recognized by antibodies elicited during acute infection. As a proof-of-principle, we applied this approach to assessing antibodies to Dengue virus (DENV). Using a panel of sera from patients with acute secondary DENV infection, we panned a DENV antigen fragment library displayed on the surface of bacteriophage MS2 virus-like particles and characterized the population of affinity-selected peptide epitopes by deep sequence analysis. Although there was considerable variation in the responses of individuals, we found several epitopes within the Envelope glycoprotein and Non-Structural Protein 1 that were commonly enriched. This report establishes a novel approach for characterizing pathogen-specific antibody responses in human sera, and has future utility in identifying novel diagnostic and vaccine targets.

Evaluation of the safety and adjuvant effect of a detoxified listeriolysin O mutant on the humoral response to dengue virus antigens.

Listeriolysin O (LLO) has been proposed as a potential carrier or adjuvant molecule in the vaccination field. However, the cytotoxic and pro-apoptotic effects of LLO are the major limitations for this purpose. Here, we have performed a preclinical safety evaluation and characterized a new potential adjuvant application for a non-cytolytic LLO mutant (dtLLO) to enhance and modulate the immune response against the envelope (E) protein from dengue virus. In addition, we have studied the adjuvant effects of dtLLO on human immune cells and the role of membrane cholesterol for the binding and proinflammatory property of the toxoid. Our in-vivo results in the murine model confirmed that dtLLO is a safer molecule than wild-type LLO (wtLLO), with a significantly increased survival rate for mice challenged with dtLLO compared with mice challenged with wtLLO (P< 0·001). Histopathological analysis showed non-toxic effects in key target organs such as brain, heart, liver, spleen, kidney and lung after challenge with dtLLO. In vitro, dtLLO retained the capacity of binding to plasma membrane cholesterol on the surface of murine and human immune cells. Immunization of 6-8-week-old female BALB/c mice with a combination of dtLLO mixed with E protein elicited a robust specific humoral response with isotype diversification of immunoglobulin (Ig)G antibodies (IgG1 and IgG2a). Finally, we demonstrated that cholesterol and lipid raft integrity are required to induce a proinflammatory response by human cells. Taken together, these findings support a potential use of the dtLLO mutant as a safe and effective adjuvant molecule in vaccination.

The Analysis of Yellow Fever Virus Antigen in Human Serum from Epidemic Areas of Tianjin Port, 2012

Objective: To investigate the prevalence and distribution characteristics of yellow fever virus (YFV) antigen in human serum from epidemic areas of Tianjin port in 2012. Methods: The people from the yellow fever epidemic areas of Tianjin port were selected as study object. 172 samples were collected together with detailed personal information. And each sample contained 5 ml venous blood. Indirect ELISA was used to detect YFV antigen. The dengue virus antigen and west nile virus antigen were also detected in positive samples to reduced cross reactivity. Positive rate was calculated. Statistical methods were used to compare the differences of the positive rates between different countries, genders, ages, occupations and entry time. Results: All respondents came from Africa and South America. The total positive rate of serum antigen of YFV was 11.63% (20/172). Of which, the positive rates of African and South American people were 10.96% and 15.38%, respectively. The positive rates of male and female were 11.68% and 11.43%, respectively. The positive rate of >40 year old age group was the highest, up to 17.24%. In the time distribution, the positive rate of third-quarter entry personnel was up to 14.94%. There was no significant difference in positive rate between different countries, genders, ages, and entry time, except occupations. Workers engaged in labor service positive rate was 29.41%. Conclusion: The YFV antigen positive rate of people from epidemic areas in 2012 was high. These people carrying pathogens pose a threat to public health security of China as a potential source of infection. There was a significant difference in the detection rate of YFV antibody among people with different occupations.

Maintenance of demographic and hematological profiles in a long-lasting dengue fever outbreak: implications for management.

BackgroundDengue fever (DF) outbreaks present regionally specific epidemiological and clinical characteristics. In certain medium-sized cities (100 000–250 000 inhabitants) of São Paulo State, Brazil, and after reaching an incidence of 150 cases/100 000 inhabitants (“epidemiological threshold”), clinical diagnosis indicated dengue virus (DENV) infection. During this period, other seasonally infectious diseases with symptoms and physical signs mimicking DF can simultaneously occur, with the consequential overcrowding of health care facilities as the principal drawbacks. Confirmation of clinical diagnosis of DF with serological tests may help in avoiding faulty diagnosis in patients, who might later undergo dengue hemorrhagic fever (DHF) and the dengue-shock syndrome (DSS). Furthermore, demographic and hematological profiles of patients are useful in detecting specific early characteristics associated to DF, DHF and DSS.MethodsFrom March to June, 2007, 456 patients from Marilia in northwest São Paulo State who had only been diagnosed for DF by clinical criteria, underwent serologic testing for non-structural 1 (NS1) DENV antigens. Individual results were used in comparative analysis according to demographic (gender, age) and hematological (leukocyte and platelet counts, percentage of atypical lymphocytes) profiles. Temporal patterns were evaluated by subdividing data according to time of initial attendance, using recorded variables as predictors of DENV infection in logistic regression models and ROC curves.ResultsSerologic DENV detection was positive in 70.6 % of the patients. Lower leukocyte and platelet counts were the most important factors in predicting DENV infection (respective medians DENV + = 3 715 cells/ml and DENV- = 6 760 cells/ml, and DENV + = 134 896 cells/ml and DENV- = 223 872 cells/ml). Furthermore, all demographic and hematological profiles presented a conservative temporal pattern throughout this long-lasting outbreak.ConclusionsAs consistency throughout the epidemic facilitated defining the conservation pattern throughout the early stages, this was useful for improving management during the remaining period.Electronic supplementary materialThe online version of this article (doi:10.1186/s40249-016-0177-y) contains supplementary material, which is available to authorized users.

Measuring Haitian children's exposure to chikungunya, dengue and malaria.

ObjectiveTo differentiate exposure to the newly introduced chikungunya virus from exposure to endemic dengue virus and other pathogens in Haiti.MethodsWe used a multiplex bead assay to detect immunoglobulin G (IgG) responses to a recombinant chikungunya virus antigen, two dengue virus-like particles and three recombinant Plasmodium falciparum antigens. Most (217) of the blood samples investigated were collected longitudinally, from each of 61 children, between 2011 and 2014 but another 127 were collected from a cross-sectional sample of children in 2014.FindingsOf the samples from the longitudinal cohort, none of the 153 collected between 2011 and 2013 but 78.7% (48/61) of those collected in 2014 were positive for IgG responses to the chikungunya virus antigen. In the cross-sectional sample, such responses were detected in 96 (75.6%) of the children and occurred at similar prevalence across all age groups. In the same sample, responses to malarial antigen were only detected in eight children (6.3%) but the prevalence of IgG responses to dengue virus antigens was 60.6% (77/127) overall and increased steadily with age. Spatial analysis indicated that the prevalence of IgG responses to the chikungunya virus and one of the dengue virus-like particles decreased as the sampling site moved away from the city of Léogâne and towards the ocean.ConclusionSerological evidence indicates that there had been a rapid and intense dissemination of chikungunya virus in Haiti. The multiplex bead assay appears to be an appropriate serological platform to monitor the seroprevalence of multiple pathogens simultaneously.

Expression and assembly of cholera toxin B subunit and domain III of dengue virus 2 envelope fusion protein in transgenic potatoes

The rates of mosquito-transmitted dengue virus infection in humans have increased in tropical and sub-tropical areas. Domain III of dengue envelope protein (EDIII) is involved in cellular receptor binding and induces serotype-specific neutralizing antibodies. EDIII fused to the B subunit of Vibrio cholera (CTB–EDIII) was expressed in potatoes to develop a plant-based vaccine against dengue virus type 2. CTB–EDIII fused to an endoplasmic reticulum (ER) retention signal, SEKDEL, was introduced into potatoes by A. tumefaciens-mediated gene transformation. The integration of the CTB–EDIII fusion gene into the nuclear genome of transgenic plants was confirmed by genomic DNA polymerase chain reaction (PCR), and mRNA transcripts of CTB–EDIII were detected. CTB–EDIII fusion protein was expressed in potato tubers and assembled into a pentameric form capable of binding monosialotetrahexosylganglioside (GM1). The level of expression was determined to be ∼0.005% of total soluble protein in potato tubers. These results suggest that dengue virus antigen could be produced in potatoes, raising the possibility that edible plants are employed in mucosal vaccines for protection against dengue infection.

Antiviral and Immunomodulatory Effects of Norantea brasiliensis Choisy on Dengue Virus-2

Background/Aims: Severe dengue fever is a result of exacerbated immune responses and no specific treatments are available. We evaluated the antiviral and immunomodulatory effects of Norantea brasiliensis Choisy. Methods: Human adherent monocytes infected in vitro with dengue virus (DENV)-2 were incubated with the crude ethanol extract from leaves (NB1) or 3 derived fractions: dichloromethane (NB3), ethyl acetate (NB5), and butanolic (NB6) partitions. The antiviral and immunomodulatory activities were determined by intracellular detection of DENV antigen within monocytes and by secreted NS1 viral protein and cytokines. Results: The crude extract alone exhibited both antiviral activities (intracellular and secreted antigens) and all fractions derived from this extract modulated NS1 production. Regarding the immunomodulatory effect, among the secreted factors, TNF-α was inhibited by NB3 and NB6; IL-6 was inhibited by NB1, NB3, and NB6; IL-10 by NB1 and NB3; and IFN-α by NB6. The crude extract (NB1) presented the best antiviral effect, whereas the dichloromethane fraction (NB3) presented an immunomodulatory effect in the inflammatory and anti-inflammatory cytokines. Conclusion: During in vitro DENV infection, N. brasiliensis Choisy exerts both antiviral and immunomodulatory effects that are likely associated, considering that less viral load may lead to less immunostimulation.

An epidemiological study of dengue in Delhi, India

Delhi, the capital of India, is an important metropolitan hub for major financial and sociocultural exchanges, offering challenging threats to current public health infrastructure. In recent past, an upsurge of dengue cases in Delhi posed a significant menace to the existing dengue control policies. To reform the control strategies and take timely intervention to prevent future epidemics, an epidemiological study on the proportion of both asymptomatic and symptomatic dengue infections in selected population was conducted. The aim of the study was to investigate and assess the epidemiology of dengue infection and to estimate the proportion of asymptomatic and symptomatic dengue infections in Delhi. In this study, around 50 confirmed dengue cases, a total of 2125 individuals as household and neighbourhood contacts, with or without dengue febrile illness, were finger pricked and serologically detected as dengue positive or negative using SD Duo Bioline Rapid Diagnostic Test (SD Inc, Korea) with NS1, IgM & IgG combo test, which detected dengue virus antigen and antibodies to dengue virus in human blood. Out of 2125 individuals, 768 (36.1%) individuals showed positive dengue test with past (25.5%), primary (1.88%) or secondary (8.8%) dengue infections. Higher percentage of IgG was found in age groups 15–24 years and 25–50 years (36% each). Infants (<1 year) presented higher incidence of new infections (22% of NS1+IgM positives) as compared to adults. Further analysis revealed that out of the 226 newly infected cases (including NS1 and IgM positives), 142 (63%) were asymptomatic and 84 (37%) were symptomatic, as per WHO guidelines. Our findings also suggest that out of the total population screened, 10.6% dengue infection was either primary or secondary. On the basis of these results, it may be hypothesized that there are large number of asymptomatic dengue infections in the community as compared to reported symptomatic cases in Delhi. For the effective control of dengue transmission in such community like Delhi where dengue epidemics have frequently been encountered, it is essential to ascertain the proportion of asymptomatic dengue infections which may act as a reservoir for dengue transmission, as well as threat for developing dengue haemorrhagic fever (DHF).

Hepatobiliary Quiz (Answers)—15 (2015)

Hepatobiliary Quiz (Answers)—15 (2015)

The immunogenicity of tetravalent dengue DNA vaccine in mice pre-exposed to Japanese encephalitis or Dengue virus antigens.

Asian countries are an endemic area for both dengue (DENV) and Japanese encephalitis viruses (JEV). While JEV vaccines have been used extensively in this region, DENV vaccines remains under development. Whether preexisting naturally acquired or vaccination-induced immunity against JEV may affect the immune response to dengue vaccine candidate is unclear. In this study we used mice previously immunized with JEV vaccines to evaluate the impact on dengue-specific neutralizing antibody responses to a tetravalent dengue DNA vaccine candidate (TDNA). A tetravalent cocktail of plasmids encoding pre-membrane and envelope proteins from each dengue serotype was administered into mice which had been previously primed with inactivated or live-attenuated JEV vaccines, or dengue serotype2 virus (DENV-2). Neutralizing antibody response was measured employing a plaque reduction neutralization test at two weeks after the priming and at four weeks after the second dose of the dengue tetravalent plasmids. Inactivated or live-attenuated JEV vaccines, or DENV-2 induced low levels of neutralizing antibodies against the homologous viruses (JE and dengue virus, respectively). DENV-2 injection induced also low levels of cross-reactive antibodies against DENV-1, -3 and -4. JEV vaccines have no effect on the dengue-specific neutralizing antibody responses to the subsequent TDNA immunization. Pre-exposure to DENV-2 infection increased DENV-2 specific response neutralizing antibody to two doses of TDNA plasmids by six folds, but did not affect antibody response to other serotypes. Priming with JEV vaccines did not impact on dengue virus-specific neutralizing antibody response to a dengue TDNA vaccine candidate in mice.

Vascular leakage in dengue hemorrhagic Fever is associated with dengue infected monocytes, monocyte activation/exhaustion, and cytokines production.

The vascular leakage was shown by the increment of hematocrit (Hct), dengue viral infected monocyte, monocyte status, and cytokines production in patients infected with dengue virus. Dengue viral antigens were demonstrated in monocytes (CD14+) from peripheral blood mononuclear cells. The increased levels of Hct, interleukin- (IL-) 10, and tumor necrosis factor-alpha (TNF-α) were detected in dengue fever (DF), dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) patients as compared with other febrile illnesses (OFIs). The highest levels of Hct and IL-10 were detected in DSS patients as compared with other groups (P < 0.05) especially on one day before and after defervescence. The unstimulated and lipopolysaccharide- (LPS-) stimulated monocytes from DSS patients showed the significantly decreased of intracellular IL-1β and TNF-α. In addition, the lowest level of mean fluorescence intensity (MFI) of CD11b expression on monocytes surface in DSS patients was also demonstrated. Furthermore, the negative correlations between IL-10 levels and intracellular IL-1β and MFI of CD11b expression in unstimulated and LPS-stimulated monocytes were also detected. Nevertheless, not only were the relationships between the prominent IL-10 and the suppression of intracellular monocyte secretion, namely, IL-1β, TNF-α, demonstrated but also the effect of vascular leakage was observed.

Comparison of E and NS1 antigens capture ELISA to detect dengue viral antigens from mosquitoes

Comparison of E and NS1 antigens capture ELISA to detect dengue viral antigens from mosquitoes

Atrial Fibrillation Due to Acute Myocarditis During Dengue Haemorrhagic Fever.

Atrial Fibrillation Due to Acute Myocarditis During Dengue Haemorrhagic Fever.

Screening of blood serum for markers of infection with measles, rubella, cytomegalovirus and Dengue Fever in Russian tourists

The aim of the study was in the differential specific diagnosis of viral infections of unknown etiology in patients who have recently returned from a trip to the countries of Southeast Asia and India. In 2013, SRC VB "Vector" examined single serum samples from 131 patients with suspected Dengue Fever. Dengue viral RNA was detected by means of polymerase chain reaction, NS dengue virus antigen and antibody classes M and G to Dengue virus - with the use of immunochromatographic methods and enzyme immunoassay. In the same sera the presence of specific IgM to measles, rubella and cytomegalovirus virus antigen was revealed by enzyme immunoassay and enzyme-enhanced chemiluminescence. As a result of examination 87 (66.4%) ofsamples were diagnosed as "Dengue fever". In seven out of the 87 samples there were detected specific class-M antibodies to cytomegalovirus. 44 sera (33.6%) out of 131 did not contain markers for Dengue fever. But 4 out of them contained IgM to measles virus, the other 5 sera contained IgM to rubella virus. Regional Centre of WHO laboratory confirmed three cases of measles, the one positive result for the content of IgM to rubella virus antigens results was interpreted as "equivalent." Identified in 2013 cases of importation ofmeasles and rubella by tourists from Southeast Asia and India have not resulted in the transmission of the disease through contact with others persons in Russia. In seven detected Dengue Fever cases opportunistic cytomegalovirus infection was revealed.

Preliminary evaluation on the efficiency of the kit Platelia Dengue NS1 Ag-ELISA to detect dengue virus in dried Aedes aegypti: a potential tool to improve dengue surveillance

BackgroundSurveillance is a critical component of any dengue prevention and control programme. Herein, we investigate the efficiency of the commercial kit Platelia Dengue NS1 Ag-ELISA to detect dengue virus (DENV) antigens in Aedes aegypti mosquitoes infected under laboratory conditions.MethodsUnder insectary conditions, four to five day-old mosquitoes were orally challenged with DENV-2 titer of 3.6 x 105 PFU equivalent/ml, incubated for 14 days and then killed. At ten time-points following mosquito death (0, 6, 12, 24, 72, 96, 120, 144 and 168 h), i.e., during a one-week period, dried mosquitoes were comparatively tested for the detection of the NS1 antigen with other methods of detection, such as qRT-PCR and virus isolation in C6/36 cells.ResultsWe first observed that the NS1 antigen was more effective in detecting DENV-2 in Ae. aegypti between 12 and 72 h after mosquito death when compared with qRT-PCR. A second round involved comparing the sensitivity of detection of the NS1 antigen and virus isolation in C6/36 cells. The NS1 antigen was also more effective than virus isolation, detecting DENV-2 at all time-points, i.e., up to 168 h after mosquito death. Meanwhile, virus isolation was successful up to 96 h after Ae. aegypti death, but the number of positive samples per time period presented a tendency to decline progressively over time. From the 43 samples positive by the virus isolation technique, 38 (88.4%) were also positive by the NS1 test.ConclusionTaken together, these results are the first to indicate that the NS1 antigen might be an interesting complementary tool to improve dengue surveillance through DENV detection in dried Ae. aegypti females.

Dengue virus localization in human tissues (case study)

Dengue is an arboviral disease which is present in tropical and subtropical regions of the world. Dengue virus (DENV) can cause benign dengue fever (DF) to the most severe forms, dengue hemorrhagic fever and dengue shock syndrome (DHF/DSS). Many studies have been confirmed the dengue viral antigens in various types of naturally infected human tissues but little work has been done and is not certain the localization of viral RNA in human tissues. We studied tissue specimens of two patients who died from clinically diagnosed DHF to confirm serologically or virologically. DENV was detected by dengue specific Reverse transcription polymerase chain reaction (RT-PCR) for the detection of dengue viral RNA. From the RT-PCR, dengue viral RNA was detected in liver, kidney and lungs tissues while the DENV RNA absence in brain, heart, spleen and pancreas tissues. DENV can infect a number of different tissue types.

Epidemiology and seropositivity of dengue fever cases in a rural tertiary care hospital of western Maharashtra, India

Introduction: Dengue fever is rapidly emerging in India, even in non endemic areas. Dengue fever is more commonly seen in adults and older children. It was earlier confined to urban areas and now has penetrated into rural setup. Clinical presentation varies from mild to severe fatal form; therefore laboratory diagnosis is very essential. Objective: To study the seropositivity of clinically suspected dengue fever cases Material and method: Blood samples from clinically suspected dengue fever cases were screened for NS1 dengue virus antigen and IgM and IgG dengue specific antibodies by rapid immuno chromatographic test (ICT) Dengue Day 1 kit (Mfd by J. Mitra & Co. Pvt. Ltd). Result: Out of total 917 blood samples tested, 281(30.6%) were positive for one or more of three markers. Of 281 blood samples NS1 was positive in 198 cases while NS1 with either IgM or IgG was positive in 16 cases. Only IgG in 25cases and only IgM in 28 cases was observed. All the three parameters were positive in 3 cases. Conclusion: From the present study it can be concluded that the dengue cases are on rise and this underlines the need for continuous surveillance and rapid implementation of dengue control programme.

C5a receptor-targeting ligand-mediated delivery of dengue virus antigen to M cells evokes antigen-specific systemic and mucosal immune responses in oral immunization

Oral mucosal immunization is a feasible and economic vaccination strategy. In order to achieve a successful oral mucosal vaccination, antigen delivery to gut immune inductive site and avoidance of oral tolerance induction should be secured. One promising approach is exploring the specific molecules expressed on the apical surfaces of M cells that have potential for antigen uptake and immune stimulation. We previously identified complement 5a receptor (C5aR) expression on human M-like cells and mouse M cells and confirmed its non-redundant role as a target receptor for antigen delivery to M cells using a model antigen. Here, we applied the OmpH ligand, which is capable of targeting the ligand-conjugated antigen to M cells to induce specific mucosal and systemic immunities against the EDIII of dengue virus (DENV). Oral immunization with the EDIII–OmpH efficiently targeted the EDIII to M cells and induced EDIII-specific immune responses comparable to those induced by co-administration of EDIII with cholera toxin (CT). Also, the enhanced responses by OmpH were characterized as Th2-skewed responses. Moreover, oral immunization using EDIII–OmpH did not induce systemic tolerance against EDIII. Collectively, we suggest that OmpH-mediated targeting of antigens to M cells could be used for an efficient oral vaccination against DENV infection.