B henselae stimulation is dependent on TLR2 but not TLR4. HEK293 cells were transiently transfected with TLR4/MD-2 or TLR4/MD-2 and CD14 (A); or TLR2, TLR2 and TLR6, TLR2 and TLR1, or TLR6 and TLR1 in the absence (B) or in the presence of CD14 (C). Cells were stimulated with various numbers of B henselae, Pam3CSK4 (P3), LPS, IL-1β, TNFα, or medium alone. Culture supernatants were harvested after 18 hours and CXCL8 levels were determined by ELISA. DCs were pretreated with 10 μg/mL anti-TLR2 or anti-TLR4 antibody before the addition of B henselae or the indicated stimuli (D). CXCL8 release was measured by ELISA. Data shown (mean ± SD of triplicates) are from 1 of 2 independent experiments. Ctrl indicates control.