Involvement of adenosine, LTB₄, and chemokines in an autocrine/paracrine mechanism of IgE- and IgE+Ag-induced mast cell migration. (A) CFSE-labeled wild-type and FcϵRIα^-/- BMMCs in upper wells were attracted to lower wells containing wild-type cells and SPE-7 or 206 IgE-sensitized wild-type cells and antigen. (B) Supernatants of wild-type BMMCs incubated with SPE-7 for the indicated periods attracted FcϵRIα^-/- cells. Supernatants of PC IgE (206)-sensitized wild-type cells incubated with antigen for the indicated periods also attracted FcϵRIα^-/- cells. (C) Migration was induced by adenosine and LTB₄, but not by sphingosine 1-phosphate (S1P) or histamine. (D) Pertussis toxin (PTX), adenosine deaminase (ADA), and adenosine A₃ receptor inhibitor MRS 1523 inhibit the migration of CFSE-labeled wild-type BMMCs from upper wells to lower wells containing IgE-sensitized wild-type cells and antigen or adenosine. (E) 5-Lipoxygenase inhibitor (N-oleoyl dopamine [ODA]), BLT₁ receptor inhibitor (U-75302), and BLT₂ inhibitor (LY2552833) inhibit the migration of CFSE-labeled wild-type BMMCs from upper wells to lower wells containing IgE-sensitized wild-type cells and antigen. U-75302 and LY2552833 at 1 μM each were used in a combination (U+LY) as well. (F) Neutralizing antibodies to several chemokines inhibit the migration of CFSE-labeled wild-type BMMCs from upper wells to lower wells containing IgE-sensitized wild-type cells and antigen. All the antibodies at the higher concentrations each were used in a combination (all) as well. Mean values ± SD are shown out of 2 independent experiments.