Figure 4

H. polygyrus bakeri-induced CD11c⁻CD11b⁺Gr1⁺ cells suppress CD4⁺ Th2 cell function in vitro. (a) CD11c⁻CD11b⁺Gr1⁺ cells were purified from mesenteric lymph node (MLN) (top two panels) and spleen cells (lower two panels) obtained from naive and infected mice on day 7 after primary infection, co-cultured with carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled OT-II spleen cells at the indicated ratios, and stimulated with 1 nM OVA_323–339 peptide. Seventy-two hours later, the cells from triplicate wells were harvested and pooled, stained with APC-conjugated anti-CD4 mAb and the CFSE dilution was analyzed by flow cytometry in gated CD4⁺ cells. Data are representative of three independent experiments. (b) IL-4 levels secreted by adult worm homogenate (AWH)-stimulated MLN and spleen cells on days 7 and 14 after primary infection. *P<0.05 for MLN cells on day 14 vs. day 7 p.i.; ****P<0.0001 for spleen cells on day 14 vs. day 7 p.i.; ND, not detected. (c) CD11c⁻CD11b⁺Gr1⁺ cells were purified from MLN and spleen cells of infected mice on day 7 after primary infection, pooled, and co-cultured with unfractionated spleen cells obtained from infected mice on day 14 p.i. at the indicated ratios. Co-cultures were stimulated with 50 μg ml⁻¹ adult worm homogenate (AWH), and IL-4 levels were determined in supernatants collected at 48 h by ELISA. ***P<0.001 compared with spleen cells without CD11c⁻CD11b⁺Gr1⁺ cells.

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