Fig. 6

T cell impact on MDM polarization state. (A) Primary MDMs were cultured in mono-culture (M0) for 6 days and then polarized to M0, M1-like, or M2-like phenotypes for 3 days followed by either maintenance as mono-culture or combined into co-culture with autologous T cell cultures for the last 24 h. (B) RNA expression was interrogated for all polarized MDM mono-culture conditions and co-culture with T cells conditions. Data expressed as normalized relative quantity (NRQ), as related to house-keeping genes RPLP0 and POLR2A. Paired Wilcoxon non-parametric test for corresponding polarization conditions; n = 6; error bar is mean with SEM. (C) Secreted cytokine and chemokine protein concentrations measured in cell culture supernatant media from polarized MDM mono-cultures and co-cultures with autologous T cells, expressed as pg/mL. Paired parametric test for corresponding polarization conditions; n = 3; Data represent mean ± SEM. (D–F) T cell migration analysis of T cell mono-culture and the three co-culture conditions was performed by in-chip confocal microscopy imaging. Data is expressed as each sample’s fold change in migration distance relative to its corresponding mono-culture condition, of (D) total T cell population and (E) CD4 or CD8 T cell sub-populations. One sample Wilcoxon signed-rank test with hypothetical value at 1 and ignoring values that match hypothetical value; n = 6; error bar is mean ± SEM. (F) Ratio of CD4 to CD8 absolute migration distances for T cell mono-culture and the MDM polarization co-culture conditions. One-way ANOVA nonparametric with Friedman post-hoc analysis comparing the mean rank of each column with the rank of every other column; n = 6; Data represent mean ± SEM. (G) T cell mRNA expression in T cell mono-culture and polarized MDM co-culture conditions. Data expressed as normalized relative quantity (NRQ) as related to house-keeping genes RPLP0 and POLR2. One-way ANOVA nonparametric with Friedman post-hoc analysis comparing the mean rank of each column with the rank of every other column; n = 5; Data represent mean ± SEM.