Dengue: A review of laboratory diagnostics in the vaccine age.

Background. Dengue is an important arboviral infection of considerable public health significance. It occurs in a wide global belt within a variety of tropical regions. The timely laboratory diagnosis of Dengue infection is critical to inform both clinical management and an appropriate public health response. Vaccination against Dengue virus is being introduced in some areas.Discussion. Appropriate diagnostic strategies will vary between laboratories depending on the available resources and skills. Diagnostic methods available include viral culture, the serological detection of Dengue-specific antibodies in using enzyme immunoassays (EIAs), microsphere immunoassays, haemagglutination inhibition or in lateral flow point of care tests. The results of antibody tests may be influenced by prior vaccination and exposure to other flaviviruses. The detection of non-structural protein 1 in serum (NS1) has improved the early diagnosis of Dengue and is available in point-of-care assays in addition to EIAs. Direct detection of viral RNA from blood by PCR is more sensitive than NS1 antigen detection but requires molecular skills and resources. An increasing variety of isothermal nucleic acid detection methods are in development. Timing of specimen collection and choice of test is critical to optimize diagnostic accuracy. Metagenomics and the direct detection by sequencing of viral RNA from blood offers the ability to rapidly type isolates for epidemiologic purposes.Conclusion. The impact of vaccination on immune response must be recognized as it will impact test interpretation and diagnostic algorithms.

[Dengue vaccines, a time for cautious optimism?] / Vaccins contre la dengue, le temps de l'optimisme prudent ?

Vaccine could take a central role in the strategy to reduce the burden of dengue. The development of an effective and safe vaccine must address various immunological challenges. Several vaccines are currently in development. To date, two live-attenuated vaccines have been deployed. Both have an effectiveness that varies depending on the serotypes. The deployment of the Dengvaxia vaccine, which began in 2015, was marked by a major safety alert leading to its use being restricted to previously dengue-seropositive people over 9 years old. The Qdenga vaccine is currently being deployed. There is for now insufficient data to ensure its safety in seronegative people. Some travelers, who have previously been infected with dengue, are a group for whom a vaccination recommendation applies.

Les vaccins pourraient occuper une place centrale dans la stratégie de réduction du fardeau de la dengue. Le développement d'un vaccin efficace et sûr est complexe car il doit relever plusieurs défis immunologiques. Différents vaccins sont en développement. À ce jour, deux vaccins vivants atténués ont été déployés. Tous deux ont une efficacité qui varie selon les sérotypes. Le déploiement du vaccin Dengvaxia, débuté en 2015, a été marqué par une alerte de sécurité majeure conduisant à restreindre son usage aux personnes de plus de 9 ans, préalablement séropositives pour la dengue. Le vaccin Qdenga est en cours de déploiement. Le recul est insuffisant pour assurer son innocuité chez les séronégatifs. Certains voyageurs, ayant déjà été infectés par la dengue, constituent un groupe pour lequel une recommandation vaccinale s'applique.

Regional Variation of the CD4 and CD8 T Cell Epitopes Conserved in Circulating Dengue Viruses and Shared with Potential Vaccine Candidates.

As dengue expands globally and many vaccines are under trials, there is a growing recognition of the need for assessing T cell immunity in addition to assessing the functions of neutralizing antibodies during these endeavors. While several dengue-specific experimentally validated T cell epitopes are known, less is understood about which of these epitopes are conserved among circulating dengue viruses and also shared by potential vaccine candidates. As India emerges as the epicenter of the dengue disease burden and vaccine trials commence in this region, we have here aligned known dengue specific T cell epitopes, reported from other parts of the world with published polyprotein sequences of 107 dengue virus isolates available from India. Of the 1305 CD4 and 584 CD8 epitopes, we found that 24% and 41%, respectively, were conserved universally, whereas 27% and 13% were absent in any viral isolates. With these data, we catalogued epitopes conserved in circulating dengue viruses from India and matched them with each of the six vaccine candidates under consideration (TV003, TDEN, DPIV, CYD-TDV, DENVax and TVDV). Similar analyses with viruses from Thailand, Brazil and Mexico revealed regional overlaps and variations in these patterns. Thus, our study provides detailed and nuanced insights into regional variation that should be considered for itemization of T cell responses during dengue natural infection and vaccine design, testing and evaluation.

Analysis of a fractional-order model for dengue transmission dynamics with quarantine and vaccination measures.

A comprehensive mathematical model is proposed to study two strains of dengue virus with saturated incidence rates and quarantine measures. Imperfect dengue vaccination is also assumed in the model. Existence, uniqueness and stability of the proposed model are proved using the results from fixed point and degree theory. Additionally, well constructed Lyapunov function candidates are also applied to prove the global stability of infection-free equilibria. It is also demonstrated that the model is generalized Ulam-Hyers stable under some appropriate conditions. The model is fitted to the real data of dengue epidemic taken from the city of Espirito Santo in Brazil. For the approximate solution of the model, a non-standard finite difference(NSFD) approach is applied. Sensitivity analysis is also carried out to show the influence of different parameters involved in the model. The behaviour of the NSFD is also assessed under different denominator functions and it is observed that the choice of the denominator function could influence the solution trajectories. Different scenario analysis are also assessed when the reproduction number is below or above one. Furthermore, simulations are also presented to assess the epidemiological impact of dengue vaccination and quarantine measures for infected individuals.

Molecular mechanisms in the pathogenesis of dengue infections.

Dengue is the most rapidly emerging climate-sensitive infection, and morbidity/mortality and disease incidence are rising markedly, leading to healthcare systems being overwhelmed. There are currently no specific treatments for dengue or prognostic markers to identify those who will progress to severe disease. Owing to an increase in the burden of illness and a change in epidemiology, many patients experience severe disease. Our limited understanding of the complex mechanisms of disease pathogenesis has significantly hampered the development of safe and effective treatments, vaccines, and biomarkers. We discuss the molecular mechanisms of dengue pathogenesis, the gaps in our knowledge, and recent advances, as well as the most crucial questions to be answered to enable the development of therapeutics, biomarkers, and vaccines.

XI Reunión Ad Hoc del Grupo Técnico Asesor (GTA) sobre Enfermedades Prevenibles por Vacunación. 21 de noviembre del 2023. Reunión virtual

La Región de las Américas está recuperando sus tasas de cobertura de inmunización para la mayoría de los antígenos. En el 2022, la tasa de cobertura de la tercera dosis de la vacuna contra la difteria, el tétanos y la tos ferina (DTP3) fue del 90%, frente al 86% en 2021. En total, 1,3 millones de niños menores de 1 año siguen sin vacunar, frente a los 1,9 millones de 2021. Por supuesto, el camino hacia la recuperación de la pandemia de COVID-19 es largo, pero las Américas están mostrando signos de progreso. En 2002, los casos de dengue superaron el millón, mientras que en 2013 se registraron más de 2 millones, y más de 3 millones en 2019. Aunque la tasa regional de letalidad por dengue se mantiene por debajo del 0,05%, el aumento de la transmisión está socavando los esfuerzos de recuperación social y económica de los países. En septiembre de 2023, el Grupo de Expertos en Asesoramiento Estratégico (SAGE) sobre Inmunización de la Organización Mundial de la Salud (OMS) recomendó el uso de la serie de dos dosis de la vacuna contra el dengue TAK-003 producida por Takeda para niños de 6 a 16 años que viven en entornos con alta carga de enfermedad por dengue y alta intensidad de transmisión. Durante esta XI reunión del Grupo Técnico Asesor (GTA), la Secretaría de la Organización Panamericana de la Salud (OPS) pidió a los miembros del GTA que consideraran la evidencia sobre la seguridad y eficacia de esta vacuna y propusieran recomendaciones para su uso en las Américas. Asimismo, el virus sincitial respiratorio (VSR) es motivo de gran preocupación en las Américas. Los datos notificados por los Estados Miembros a la red integrada de vigilancia respiratoria SARInet Plus de la OPS indican que el VSR contribuye significativamente a la carga de enfermedades respiratorias en la Región. Por grupos de edad, los casos y hospitalizaciones asociados al VSR se han notificado principalmente entre lactantes menores de 2 años. En los últimos meses, tanto la Administración de Alimentos y Medicamentos de EE.UU. (FDA) como la Agencia Europea del Medicamento (EMA) han aprobado la vacuna Abrysvo contra el VRSpreF producida por Pfizer para mujeres embarazadas, con el objetivo de reducir la incidencia del VRS entre los recién nacidos menores de 6 meses. Una vez más, los miembros del GAT fueron convocados para proporcionar sus recomendaciones a la OPS sobre el uso de esta vacuna en las Américas.

Atenção primária à saúde: a maior aliada na resposta à epidemia da dengue no Brasil

Em resposta ao cenário epidemiológico crítico de incidên- cia de casos, hospitalizações e óbitos por dengue, o Ministério da Saúde do Brasil incorporou, ainda em dezembro de 2023, a vacina contra a dengue no Calendário Nacional de Vacinação. Inicialmente, a vacina foi incorporada para crianças e adolescen- tes de 10 a 14 anos (1), faixa etária que concentra o maior número de hospitalizações pela doença depois das pessoas idosas — para quem, no entanto, a vacina não foi liberada pela Agência Nacional de Vigilância Sanitária (Anvisa). Nesse sentindo, o Brasil tornou-se o primeiro país do mundo a disponibilizar a vacina contra a dengue de forma gratuita no serviço público de saúde, juntamente com diversos métodos de controle vetorial.

A tetravalent dengue virus-like particle vaccine induces high levels of neutralizing antibodies and reduces dengue replication in non-human primates.

Dengue virus (DENV) represents a significant global health burden, with 50% of the world's population at risk of infection, and there is an urgent need for next-generation vaccines. Virus-like particle (VLP)-based vaccines, which mimic the antigenic structure of the virus but lack the viral genome, are an attractive approach. Here, we describe a dengue VLP (DENVLP) vaccine which generates a neutralizing antibody response against all four DENV serotypes in 100% of immunized non-human primates for up to 1 year. Additionally, DENVLP vaccination produced no ADE response against any of four DENV serotypes in vitro. DENVLP vaccination reduces viral replication in a non-human primate challenge model. We also show that transfer of purified IgG from immunized monkeys into immunodeficient mice protects against subsequent lethal DENV challenge, indicating a humoral mechanism of protection. These results indicate that this DENVLP vaccine is immunogenic and can be considered for clinical evaluation. Immunization of non-human primates with a tetravalent DENVLP vaccine induces high levels of neutralizing antibodies and reduces the severity of infection for all four dengue serotypes.IMPORTANCEDengue is a viral disease that infects nearly 400 million people worldwide and causes dengue hemorrhagic fever, which is responsible for 10,000 deaths each year. Currently, there is no therapeutic drug licensed to treat dengue infection, which makes the development of an effective vaccine essential. Virus-like particles (VLPs) are a safe and highly immunogenic platform that can be used in young children, immunocompromised individuals, as well as healthy adults. In this study, we describe the development of a dengue VLP vaccine and demonstrate that it induces a robust immune response against the dengue virus for over 1 year in monkeys. The immunity induced by this vaccine reduced live dengue infection in both murine and non-human primate models. These results indicate that our dengue VLP vaccine is a promising vaccine candidate.

Low-dose dengue virus 3 human challenge model: a phase 1 open-label study.

Dengue human infection models present an opportunity to explore the potential of a vaccine, anti-viral or immuno-compound for clinical benefit in a controlled setting. Here we report the outcome of a phase 1 open-label assessment of a low-dose dengue virus 3 (DENV-3) challenge model (NCT04298138), in which nine participants received a subcutaneous inoculation with 0.5 ml of a 1.4 × 103 plaque-forming unit per ml suspension of the attenuated DENV-3 strain CH53489. The primary and secondary endpoints of the study were to assess the safety of this DENV-3 strain in healthy flavivirus-seronegative individuals. All participants developed RNAaemia within 7 days after inoculation with peak titre ranging from 3.13 × 104 to 7.02 × 108 genome equivalents per ml. Solicited symptoms such as fever and rash, clinical laboratory abnormalities such as lymphopenia and thrombocytopenia, and self-reported symptoms such as myalgia were consistent with mild-to-moderate dengue in all volunteers. DENV-3-specific seroconversion and memory T cell responses were observed within 14 days after inoculation as assessed by enzyme-linked immunosorbent assay and interferon-gamma-based enzyme-linked immunospot. RNA sequencing and serum cytokine analysis revealed anti-viral responses that overlapped with the period of viraemia. The magnitude and frequency of clinical and immunologic endpoints correlated with an individual's peak viral titre.

Perceptions of dengue risk and acceptability of a dengue vaccine in residents of Puerto Rico.

Dengvaxia is the first dengue vaccine recommended in the United States (U.S.). It is recommended for children aged 9-16 y with laboratory-confirmed previous dengue infection and living in areas where dengue is endemic. We conducted focus groups with parents and in-depth interviews with key informants (i.e. practicing pediatricians, physicians from immunization clinics, university researchers, and school officials) in Puerto Rico (P.R.) to examine acceptability, barriers, and motivators to vaccinate with Dengvaxia. We also carried out informal meetings and semi-structured interviews to evaluate key messages and educational materials with pediatricians and parents. Barriers to vaccination included lack of information, distrust toward new vaccines, vaccine side effects and risks, and high cost of/lack of insurance coverage for laboratory tests and vaccines. Motivators included clear information about the vaccine, a desire to prevent future dengue infections, the experience of a previous dengue infection or awareness of dengue fatality, vaccine and laboratory tests covered by health insurance, availability of rapid test results and vaccine appointments. School officials and parents agreed parents would pay a deductible of $5-20 for Dengvaxia. For vaccine information dissemination, parents preferred an educational campaign through traditional media and social media, and one-on-one counseling of parents by healthcare providers. Education about this vaccine to healthcare providers will help them answer parents' questions. Dengvaxia acceptability in P.R. will increase by addressing motivators and barriers to vaccination and by disseminating vaccine information in plain language through spokespersons from health institutions in P.R.

A systematic review on malaria and dengue vaccines for the effective management of these mosquito borne diseases: Improving public health.

Insect vector-borne diseases (VBDs) pose significant global health challenges, particularly in tropical and subtropical regions. The WHO has launched the "Global Vector Control Response (GVCR) 2017-2030" to address these diseases, emphasizing a comprehensive approach to vector control. This systematic review investigates the potential of malaria and dengue vaccines in controlling mosquito-borne VBDs, aiming to alleviate disease burdens and enhance public health. Following PRISMA 2020 guidelines, the review incorporated 39 new studies out of 934 identified records. It encompasses various studies assessing malaria and dengue vaccines, emphasizing the significance of vaccination as a preventive measure. The findings indicate variations in vaccine efficacy, duration of protection, and safety considerations for each disease, influencing public health strategies. The review underscores the urgent need for vaccines to combat the increasing burden of VBDs like malaria and dengue, advocating for ongoing research and investment in vaccine development.

Next-generation vaccines for tropical infectious diseases.

Tropical infectious diseases inflict an unacceptable burden of disease on humans living in developing countries. Although anti-pathogenic drugs have been widely used, they carry a constant threat of selecting for resistance. Vaccines offer a promising means by which to enhance the global control of tropical infectious diseases; however, these have been difficult to develop, mostly because of the complex nature of the pathogen lifecycles. Here, we present recently developed vaccine candidates for five tropical infectious diseases in the form of a catalog that have either entered clinical trials or have been licensed for use. We deliberate on recently licensed dengue vaccines, provide evidence why combination vaccination could have a synergistic impact on schistosomiasis, critically appraise the value of typhoid conjugate vaccines, and discuss the potential of vaccines in the efforts to eliminate vivax malaria and hookworms.

Brazil is hoping and waiting for a new vaccine as dengue rages.

A locally produced vaccine did well in a phase 3 clinical trial but won't be available until at least 2025.

Approaches of dengue control: vaccine strategies and future aspects.

Dengue, caused by the dengue virus (DENV), affects millions of people worldwide every year. This virus has two distinct life cycles, one in the human and another in the mosquito, and both cycles are crucial to be controlled. To control the vector of DENV, the mosquito Aedes aegypti, scientists employed many techniques, which were later proved ineffective and harmful in many ways. Consequently, the attention shifted to the development of a vaccine; researchers have targeted the E protein, a surface protein of the virus and the NS1 protein, an extracellular protein. There are several types of vaccines developed so far, such as live attenuated vaccines, recombinant subunit vaccines, inactivated virus vaccines, viral vectored vaccines, DNA vaccines, and mRNA vaccines. Along with these, scientists are exploring new strategies of developing improved version of the vaccine by employing recombinant DNA plasmid against NS1 and also aiming to prevent the infection by blocking the DENV life cycle inside the mosquitoes. Here, we discussed the aspects of research in the field of vaccines until now and identified some prospects for future vaccine developments.