Cross-linking of LIR9m on the surfaces of monocytes induces calcium mobilization and cytokine secretion.
(A) Monocytes from 2 different donors were labeled with Fluo-4 and then incubated with anti-LIR9, anti-LIR7, an isotype-matched control antibody, or no antibody. Calcium mobilization was measured on a fluorometric plate reader as a response to cross-linking of the primary antibodies with donkey antimouse F(ab')2 fragments. Data shown are representative of 2 of 4 different donors. Among the 4 donors, the average -fold changes and standard deviations in calcium flux relative to the nonantibody control were the following: control antibody, 0.91 ± 0.07; anti-LIR7, 3.10 ± 0.79; and anti-LIR9, 5.54 ± 1.57. (B) Monocytes were incubated on a 96-well tray coated with the antibodies against LIR1, LIR2, LIR7, LIR9, isotype-matched control antibody, or no antibody. Two days later, conditioned media were harvested and cytokine levels were measured using the Luminex assay system as described in “Materials and methods.” Data show the averages and standard deviations of 4 replicates of each sample. Data shown are representative of 3 experiments on different donors.
