Figure 7.

Functional and Cell Surface Expression Characterization of Immune Cell Populations from Reovirus-Treated versus Untreated Mice

TRAMP-C2 tumors harvested 5 days after intratumoral reovirus or PBS treatment were subjected to both mechanical and enzymatic dissociation using the gentleMACS Dissociator, while spleens harvested from the same mice were mechanically disrupted to obtain splenocyte cell suspensions. (A–D) The resulting tumor single-cell suspensions and/or splenocytes were then immuno-phenotyped to characterize (A) the different lymphoid immune populations for their functional ability to produce cytokines upon in vitro stimulation and for their expression of (B) FLT3LG, (C and D) PD-1, BTLA, and HVEM by (C) lymphocytes (CD8⁺, CD4⁺, and NK cells) and (D) innate inflammatory cell subsets (macrophages [F4/80⁺], inflammatory monocytes [Ly6Chi, CD11b⁺], neutrophils [Ly6G⁺, CD11b⁺], mature DCs [CD11c⁺, MHC II⁺], and immature DCs [CD11c⁺, MHC class II⁻]). Significant differences between immune cell populations derived from untreated or reovirus-infected tumors was determined using a two-way ANOVA; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.