Phenotypic characteristics of myeloid (A) and lymphoid cells (B) in spleen.

<p>Spleen cells from mice treated with LOS and/or PmB were incubated with CD11b-PerCP-Cy5.5, B220-APC, Ly6G-APC-Cy7, Ly6C-PE, MHCII-FITC or anti-CD4-APC, anti- CD8-PE-Cy7, anti-CD49b-PE and anti-CD19-FITC. Phenotype analysis was carried out using the Becton Dickinson FACSCalibur apparatus with Cell Quest Software. To calculate the mean, three mice of each group were tested. The percentages of myeloid cells in healthy mice were: CD11b⁺—3.50%, monocytic cells among CD11b⁺ cells—30.50%, granulocytic cells among CD11b⁺ cells—30.90%. The percentages of lymphoid cells in healthy mice were: CD4⁺—17.43%, CD8⁺ 9.26%- CD19⁺—61.67%, CD49b⁺—8.00%. Cytokine production by spleen cells after ConA (C) or LOS (D) stimulation. The concentration of IL-6, IFN-γ and IL-10 was measured in supernatants collected from above splenocytes (2x10⁶ cells/ml) incubated for 48h with ConA (0.5 μg/ml) or LOS (1 μg/ml), using commercially available ELISA kits (BD Bioscience or eBioscience). To calculate the mean, three mice of each group were analyzed. Cells from healthy mice stimulated with ConA produced 1.07 ng/ml of IFN-γ, 0.10 ng/ml IL-6 and 0,23 ng/ml of IL-10. Cells from healthy mice stimulated with LOS produced 0.01 ng/ml of IFN-γ, 0.33 ng/ml IL-6 and 3.52 ng/ml IL-10.