Figure 1.

Human primary astrocytes are permissive to CHIKV, MAYV, OROV, and ZIKV infection. Human primary astrocytes were isolated from brain surgery and infected with CHIKV, MAYV, OROV, or ZIKV at MOI 1 in different time points. (A) Immunofluorescence microscopy showing astrocytes specific cell markers: GFAP, Glial Fibrillary Acidic Protein, and GLT1, Glutamate Transporter 1 (both in red); GS, Glutamine Synthetase, and GLAST, Glutamate Aspartate Transporter 1 (both in green). Nuclei are stained with DAPI (cyan). Scale bar = 50 µm. (B) Cell viability post infection was measured at indicated time points. All infected cells were normalized by mock (uninfected) cells. (C) Virus infectivity was measured by flow cytometry using primary antibodies for virus proteins: CHIKV (J31F, anti-E1 antibody); MAYV (anti-alphavirus E1 antibody); OROV (anti-OROV serum), and ZIKV (anti-NS1 antibody) and plotted as percentage of infected cells in gated population. 10.000 events were recorded. (D) Viral production measured by plaque assay from the supernatants of infected cells. Viral titers were plotted as log₁₀ Plaque Forming Units/ml. All the experiments were performed on three replicates from two independent experiments presented here as media and standard deviation (SD).