The PAGODAS protocol: pediatric assessment group of dengue and Aedes saliva protocol to investigate vector-borne determinants of Aedes-transmitted arboviral infections in Cambodia

Jessica E Manning, Fabiano Oliveira, Daniel M Parker, Chanaki Amaratunga, Dara Kong, Somnang Man, Sokunthea Sreng, Sreyngim Lay, Kimsour Nang, Soun Kimsan, Ly Sokha, Shaden Kamhawi, Michael P Fay, Seila Suon, Parker Ruhl, Hans Ackerman, Rekol Huy, Thomas E Wellems, Jesus G Valenzuela, Rithea Leang, Jessica E Manning, Fabiano Oliveira, Daniel M Parker, Chanaki Amaratunga, Dara Kong, Somnang Man, Sokunthea Sreng, Sreyngim Lay, Kimsour Nang, Soun Kimsan, Ly Sokha, Shaden Kamhawi, Michael P Fay, Seila Suon, Parker Ruhl, Hans Ackerman, Rekol Huy, Thomas E Wellems, Jesus G Valenzuela, Rithea Leang

Abstract

Background: Mosquito-borne arboviruses, like dengue virus, continue to cause significant global morbidity and mortality, particularly in Southeast Asia. When the infectious mosquitoes probe into human skin for a blood meal, they deposit saliva containing a myriad of pharmacologically active compounds, some of which alter the immune response and influence host receptivity to infection, and consequently, the establishment of the virus. Previous reports have highlighted the complexity of mosquito vector-derived factors and immunity in the success of infection. Cumulative evidence from animal models and limited data from humans have identified various vector-derived components, including salivary components, that are co-delivered with the pathogen and play an important role in the dissemination of infection. Much about the roles and effects of these vector-derived factors remain to be discovered.

Methods/design: We describe a longitudinal, pagoda (community)-based pediatric cohort study to evaluate the burden of dengue virus infection and document the immune responses to salivary proteins of Aedes aegypti, the mosquito vector of dengue, Zika, and chikungunya viruses. The study includes community-based seroprevalence assessments in the peri-urban town of Chbar Mon in Kampong Speu Province, Cambodia. The study aims to recruit 771 children between the ages of 2 and 9 years for a three year period of longitudinal follow-up, including twice per year (rainy and dry season) serosurveillance for dengue seroconversion and Ae. aegypti salivary gland homogenate antibody intensity determinations by ELISA assays. Diagnostic tests for acute dengue, Zika and chikungunya viral infections will be performed by RT-PCR.

Discussion: This study will serve as a foundation for further understanding of mosquito saliva immunity and its impact on Aedes-transmitted arboviral diseases endemic to Cambodia.

Trial registration: NCT03534245 registered on 23 May 2018.

Keywords: Arbovirus; Cohort; Mosquito; Saliva; Seroprevalence; Vaccines; Vector.

Conflict of interest statement

Ethics approval and consent to participate

This study protocol and informed consent form in both English and Khmer were reviewed and approved by the NIAID Institutional Review Board and the Cambodian National Ethics Committee on Human Research (FWA #10451/IRB #3143). Eligible participants have an equal opportunity to join the research. Information sessions are held in advance of a pagoda community’s designated recruitment week. On arrival to the pagoda, participants and their guardians receive an additional “picture book” containing detailed information about the research in Khmer text with pictorial aids to enhance understanding and support decision making. For children and their guardians who wish to participate, a signed written formal consent in Khmer is collected by a study team physician and witness after all questions are answered. Collected data will be presented in an aggregated form; respondents’ identity will remain anonymous in any publication generated from this study.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Primary study area. The inset map (bottom right) indicates the location of Chbar Mon town, Kampong Speu (red cross) in Cambodia. The main map indicates the study area (with a green outline) and recruitment sites (green triangles) where participants are recruited for immunological indicators of mosquito exposure

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