TLR agonists affect PBMCs. (A). Human PBMCs were cultured in the absence (serum-free medium (SFM) control) or the presence of 0.89 μg/ml TLR2 agonists Pam3CSK4, 8.9 × 107 cells/ml heat-killed Listeria monocytogenes (HKLM), 1.79 μg/ml lipotechoic acid (LTA), or 8.9 μg/ml lipopolysaccharides from Porphyromonas gingivalis (PG-LPS), 8.9 μg/ml TLR3 agonist Poly (I:C), 0.89 μg/ml TLR4 agonist LPS, 0.89 μg/ml TLR5 agonist flagellin, 0.89 μg/ml TLR2/6 agonist FSL-1, 0.89 μg/ml TLR7 agonist IMIQ, 0.89 μg/ml TLR8 agonist ssRNA, 4.47 μM TLR9 agonist ODN 2006 or 1.79 μg/ml nucleotide oligomerization domain (NOD)-like receptor (NLR) agonist peptidoglycan (PGN). On day 3, the conditioned media were tested for the presence of tumor necrosis factor (TNF)-α using enzyme-linked immunosorbent assay. The results are means ± SEM of TNF-α concentration (n = 3 or more separate experiments). (B) Human PBMCs were cultured in the presence or absence of Poly (I:C), IMIQ and PGN as described for Figure 3A. After 1 day, PBMCs were removed and stained for CD86 (n = 3 separate experiments). (C) Human PBMCs were cultured in the presence or absence of 8.9 μg/ml TLR3 agonist Poly (I:C), 0.89 μg/ml TLR7 agonist IMIQ or 1.79 μg/ml NLR agonist PGN. On day 3, PBMCs were removed from the tissue culture plate and stained for major histocompatibility complex (MHC) class II. The results are means ± SEM of positive MHC class II cells (n = 4 separate experiments). *P < 0.05 and **P < 0.01 compared to no-agonist control (SFM) according to t-test.