Modulation of anti-CD3-induced IFN-γ production by TLR-stimulated macrophage supernatants. Whole A/J (A) or C57BL/6 (B) spleen cells were plated in the presence of 1.0 µg/mL of anti-CD3 antibody.
Notes: The activation was performed in the presence of 100% supernatants of macrophages stimulated with OK-432 (10 µg/mL), CpG (3 µg/mL), LPS (10 µg/mL), or Poly I:C (50 µg/mL), as indicated. Concentrations of combination of OK-432 and Poly I:C were also 10 µg/mL and 50 µg/mL, respectively. Not applicable (NA) represents splenocytes plated with anti-CD3 antibody alone in the absence of any macrophage supernatant. Cytokine levels were measured by ELISA 48 hours after stimulation. The differences between cultures containing supernatants from untreated and treated macrophages are statistically significant (***P < 0.001).
Abbreviations: TLR, Toll like receptor; TNF, Tumor Necrosis Factor; IL-10, Interleukin 10; ELISA, Enzyme linked Immunosorbent Assay; Poly I:C, Polyinosinic:polycytidylic acid; CpG, Cytosine-phosphate-Guanin; LPS, Lipopolysaccharide; IFN, Interferon.