FIG 5.

Effects of B cell depletion on granuloma cytokine and neutrophil profiles. (A) Single cell suspensions from saline control granulomas were stained for B cell markers and cytokine antibodies to determine the cytokine secretion profile of B cells via intracellular FACS. Six cytokines were selected to be assayed: IL-2, IL-6, IL-10, IL-17, TNF, and IFN-γ. Each symbol is a granuloma, and colors denote granulomas from individual animals. For IL-6, IL-10, TNF, and IFN-γ, n = 8 animals and n = 27 granulomas, and thus there were ∼4 granulomas per animal. For IL-2 and IL-17, n = 5 animals and n = 22 granulomas, and thus there were ∼4 granulomas per animal. (B) ELISAs to quantify cytokine amounts were performed on granuloma homogenates for four cytokines, IL-6, IL-10, IL-17, and TNF to determine whether B cell depletion via rituximab perturbed the cytokine balance within the granulomas. Each symbol is a granuloma (n = 16 animals, n = 136 granulomas, ∼8 granulomas per animal). (C) Neutrophil amounts were quantified based on the amount of calprotectin present within granuloma homogenates. Each symbol is a granuloma (n = 16 animals, n = 155 granulomas, ∼10 granulomas per animal). All statistical P values were obtained using the Mann-Whitney test.