Figure 7. IL-4 expression increases and IFNγ expression decreases in polarized T_H1 trog⁺ cells after removal of APC.

In vitro polarized T_H1 and T_H2 CD4⁺ T cells were used in a standard in vitro trogocytosis assay. In parallel, aliquots of polarized cells were stimulated with plate-bound anti-CD3 + anti-CD28 Abs. Fluorescence intensity of IL-4 vs. IFNγ is displayed in representative 2D scatter plots at 5 hrs. (left), 24 hrs., (2^nd from left), 48 hrs. (2^nd from right), and 72 hrs. (right)for (A) anti-CD3 + anti-CD28 stimulated T_H1 polarized cells, (B) T_H1 polarized trog⁺ cells (C) anti-CD3 + anti-CD28 stimulated T_H2 polarized, and (D) T_H2 polarized trog⁺ cells. Numbers represent frequency of cells in each quadrant. (E) Representative histograms showing intensity of GATA-3 (top) and T-bet (bottom) expression in T_H1 polarized trog⁺(thick black line) and trog⁻ (thin black line) cells, compared to anti-CD3/CD28 stimulated blasts (shaded grey), at 5 (left), and 72 (right), hours post-recovery. (F) Mean frequency of T-bet⁺ GATA-3⁻ (left), T-bet⁻ GATA-3⁺ (middle), and T-bet⁺ GATA-3⁺ (right), in vitro polarized T_H1 trog⁺ (black), trog⁻ (grey outline), and anti-CD3/CD28 stimulated cells (shaded grey) at 5 (top), and 72 (bottom) hours post-recovery. Error bars represent ±SEM from three independent experiments, * = p≤ 0.05, ** = p≤ 0.01, *** = p≤ 0.001, and **** = p≤ 0.0001. All data is representative of three independent experiments.