Healthy control_p
Blood samples were obtained from patients diagnosed with dengue virus (DENV) infection admitted to Varjira, and Thasongyang Hospitals with informed consent. We obtained the peripheral blood mononuclear cell (PBMC) samples from two Thai male adult donors, both diagnosed with DENV-4 infection, with DF and DHF severities and the viral load of 1.76*10^6 and 1.77 * 10^7, respectively. The PBMC samples were collected at the defervescence day as well as  two and one days before defervescence, and two weeks after defervescence. The samples were isolated from the heparinized blood samples using Ficoll-gradient centrifugation. Then, the samples were washed with 2% FBS RPMI and stored in 90%FBS DMSO in liquid nitrogen until used. Frozen PBMCs were thawed, resuspended, washed before dead cell removal. Cell suspension was washed again by 1X PBS and resuspended with 0.04% BSA/PBS to achieve the cell concentration of 500 cell/ul, before passing through 40 uM cell strainers twice. For the PBMC of healthy control, the sample was prepared following the same protocols as the samples from the dengue patients. The nucleic acids were extracted using Chromium 3' Single-cell Controller V2, following the manufacturers' instructions (10x Genomics, v2 chemistry). Single-cell libraries were constructed using Chromium 3' Single-cell Controller V2 following the provider's protocol with the expected cell numbers of 5,000 cells per sample. Each cDNA library was tagged with the i7 index system before being quality checked using Agilent High sensitivity D1000 Tapestation and Qubid.