Flow cytometry analysis of liver granuloma during acute schistosomiasis.

<p>BALB/c mice were infected with 100 <i>S. mansoni</i> cercariae, killed 8 weeks later and liver granuloma-associated leukocytes isolated before 4-colour flow cytometry analysis. (A) Gating strategy for analysis of surface expression of CD11b and I-A/I-E on SSC^high and live (7-AAD⁻) liver granuloma-associated leukocytes. Outlined regions define CD11b⁺I-A/I-E⁻ and CD11b⁺I-A/I-E^high, respectively. (B) CD11b⁺I-A/I-E⁻, and CD11b⁺I-A/I-E^high gated populations in A were analyzed for F4/80, Gr-1, Siglec-F, CD204, MMR, Dectin-1, CD68, CD80, CD86 and IL-4Rα expression, respectively. Greyscale histograms show relevant isotype control. (C) 4-colour flow cytometry analysis of liver granuloma-associated leukocytes gated on CD11b⁺I-A/I-E⁻ cells as described in A. F4/80 and Gr-1 double staining contour plot is shown. Outlined regions were sorted for cytospin and morphological analysis. (D) 4-colour flow cytometry analysis of liver granuloma-associated leukocytes gated on CD11b⁺F4/80⁺ cells as described in A. MHC-II (I-A/I-E) and scavenger receptor-A (CD204) double staining contour plot is shown. Outlined region was sorted for cytospin and morphological analysis. Data are representative of three independent experiments with similar results.