Transcriptional profile regulated by 1,25-dihydroxyvitamin D (Calcitriol) in primary Human Monocyte-Derived Macrophages infected by zika virus

ZIKA virus (ZIKV) infection is characterized by alterations in gene expression profile on host cells that consequently lead to an immune response. Here, we used RNA sequencing to analyze the mRNA and miRNA expression profile in human monocyte-derived macrophages (MDMs) differenitated in presenece of 1,25-dihydroxyvitamin D (Calcitriol) and infected with a Zika virus strain COL345Si-Asian at MOI of 5 and analized 24 hpi. Human peripheral blood mononuclear cells (PBMCs) from blood samples of 4 healthy donors mixed with 2% EDTA, were isolated through density gradient with Lymphoprep. Monocytes-derived Macrophages (MDM) were obtained from total PBMCs by adherence to the plastic and differentiation to macrophages was allowed to occur for 6 days in RPMI-1640 medium supplemented with 10% (v/v) FBS, 4 mM L-glutamine, 0.3% (v/v) NaCO3, and 1% (v/v) antibiotic-antimycotic solution 100X (complete medium), at 37 °C and 5% CO2, without growth factors. Additionally, MDMs were differentiated in presence of 1,25 di-hydroxyvitamin D3 to obtain D3-MDM. 1,25 di-hydroxyvitamin D3 was added to the culture media at a final concentration of 1 nM and replenished every 24 h for six days. MDM and D3-MDM monolayers were washed with warm PBS and were infected with Zika virus strain COL345Si-Asian at MOI of 5. Three hpi, the cells were washed with warm PBS to remove unbound virus. Then 24 hpi, macrophage monolayers were used for sample collection.