The goal of this research study is to help the investigators better understand serious dengue disease. The investigators will collect clinical information and up to four blood samples from participants with dengue, sepsis, or healthy people. The investigators will perform multiple analyses on the blood samples and compare results between dengue patients and those with sepsis and healthy people.
This is an observational study enrolling n=200 hospitalized individuals with dengue-like illness, n=30 individuals with sepsis, and n=10 healthy controls to evaluate immune responses in plasma leak syndromes. Sepsis and dengue patients will have blood collected at enrollment, day 1, and day 3, and dengue patients will have an additional blood draw at day 21. Healthy participants will have a single blood draw at enrollment. The blood collected will enable the evaluation of immune kinetics, including changes in neutrophils, cytokines, and adaptive responses, as dengue progresses or improves. Overall, this study will facilitate the evaluation of an understudied cell type and will provide unique data for understanding the temporal changes in the immune response to dengue virus. A better understanding of dengue pathophysiology will help identify potential therapeutic targets to improve outcomes in this disease.
Study Type
OBSERVATIONAL
Enrollment
240
Kampong Speu District Referral Hospital
Kampong Speu, Cambodia
RECRUITINGInternational Center of Excellence in Research Cambodia
Phnom Penh, Cambodia
RECRUITINGPreah Kossamak Hospital
Phnom Penh, Cambodia
RECRUITINGPercentage of low density neutrophils in mild/moderate vs severe dengue
Low density neutrophils will be isolated from the PBMC layer of centrifuged blood and characterized via flow cytometry. The percentages of low density neutrophils will be compared between dengue groups.
Time frame: From enrollment to study day 3
Mean florescence intensity of neutrophils expressing spleen tyrosine kinase
The mean florescence intensity (MFI) of neutrophils expressing spleen tyrosine kinase will be measured via flow cytometry and compared between mild/moderate vs severe dengue.
Time frame: From enrollment to study day 3.
Serum level of myeloperoxidase in mild/moderate versus severe dengue.
The serum level of myeloperoxidase will be measured in pg/mL and compared between participants with mild/moderate versus severe dengue.
Time frame: Enrollment to study day 3
Percentage of low density neutrophils in dengue versus sepsis
Low density neutrophils will be isolated from the PBMC layer of centrifuged blood and characterized via flow cytometry. The percentage of low density neutrophils will be compared in dengue versus sepsis.
Time frame: From enrollment to study day 3
Mean florescence intensity of neutrophils expressing spleen tyrosine kinase in dengue versus sepsis
Mean florescence intensity (MFI) of neutrophils expressing spleen tyrosine kinase will be measured via flow cytometry and compared between those with dengue versus sepsis. Other markers of neutrophil maturation, activation, and degranulation will be measured by flow cytometry and compared among dengue groups and between dengue and sepsis.
Time frame: From enrollment to study day 3.
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Serum level of myeloperoxidase in dengue versus sepsis
Serum level of myeloperoxidase will be measured in pg/mL and compared between participants with dengue versus sepsis. Other biomarkers of neutrophil activation such as calprotectin, resistin, and lactoferrin will be measured and compared among groups.
Time frame: Enrollment to study day 3
Serum levels of interferon-gamma and interleukin-10 (pg/mL)
Serum levels of interferon-gamma and interleukin-10 (pg/mL) will be measured and compared between dengue groups and sepsis overtime. Other cytokines may also be measured and compared among groups.
Time frame: Enrollment through day 3.
Percentage of DENV-specific activated T cells
Activation induced marker assays will be used to compare dengue virus (DENV)-specific T cell activation over time in mild/moderate versus severe dengue.
Time frame: Enrollment through day 21.
DENV1-4 geometric mean titer (GMT)
DENV1-4 GMT overtime will be compared between mild/moderate and severe dengue groups. These will also be correlated with disease outcome and GMT at convalescence.
Time frame: Enrollment to day 21
Peak dengue virus enhancing titer
Peak dengue virus enhancing titer overtime will be compared between mild/moderate and severe dengue groups. These will also be correlated with disease outcome and GMT at convalescence.
Time frame: Enrollment to day 21
Mean fluorescence intensity of specific lymphocyte populations among groups
Mean fluorescence intensity of specific lymphocyte populations will be compared mild/moderate versus severe dengue and dengue versus sepsis.
Time frame: Enrollment to day 3
Number of differentially expressed genes among groups
The number of differentially expressed genes will be compared between mild/moderate and severe dengue and dengue versus sepsis based on thresholds for statistical significance (e.g., adjusted p-value \< 0.05) and magnitude of change (e.g., absolute log2 fold change \> 1).
Time frame: Enrollment to day 3