Figure 4.

Canine Cytokine Profile Generated by Brain Tumors in the Presence of ZIKV^BR in Both In Vivo and In Vitro Models

(A–C) Cytokine profile in the serum and CSF of Pitbull (A), Boxer (B), and Dachshund (C) dogs at different time points after ZIKV^BR injection. (D) Schematic representation of canine monocyte and D-GBM non-contact co-culture assay. (E–L) Cytokines concentration in non-contact co-culture groups normalized by respective day mock condition. (M) Schematic representation of canine monocyte and D-GBM contact co-culture. (∗p < 0.05, t test in comparison with correspondent MOCK group. Two technical replicas for each two biologic replica). (N–P) Evaluation of monocytes activation with CD80 (N), CD83 (O), and CD64 (P) positive cell quantification by flow cytometer. Gating strategy for the evaluation of canine monocytes in the co-culture assay detailed in Figure S5. (Q) qRT-PCR analysis of Boxer and Dachshund tumor tissue for lymphocyte T cells (MHC, GRANZYME B, and PD1), tumor cell receptor PDL-1, inflammatory monocyte (CCR2), microglia (CX3CR1), macrophage M1 (CXCL10, CCR7, GATA3, STAT1, NOS2, and IDO1), macrophage M2 (ARG1 and SOC3), and NK cells (NKP30 and NK2D), normalized by endogenous expression (Beta-Actin).