Figure 3.

Mixed lymphocyte reaction (MLR) responses using control vs. shMyD88-transduced antigen-presenting cells. Primary human CD4⁺ T cells were isolated from buffy coats and activated by incubation with decreasing concentrations of shCnt, or shMyD88 THP-1 cells, previously differentiated with PMA, or PMA with the addition of Pam3csk4 + IFNγ. Cytokine production and proliferation were determined after 6 days of incubation. (A,B) IFNγ production, (C,D) IL-17 production, and (E,F) proliferation. For panels (A–D), one representative experiment of three performed with similar results, is shown. For proliferation (E,F), the combined results of the three independent experiments are shown. In the proliferation studies (E,F), the ARU of T cells without THP-1 cells was close to 0. (G,H) Flow cytometry analysis of intracellular IL-4 expression is presented for primary human CD4⁺ T cells stimulated by incubation with shCnt, or shMyD88 THP-1 cells, that were differentiated with PMA, or PMA with the addition Pam3csk4 + IFNγ: (G) shCnt and (H) shMyD88 cells. One representative experiment of three independent MLR experiments is shown in the dot plots, and the average of the three experiments is shown in the bar graphs. The two-tailed t-test was used for statistical comparison.