Figure 2.

PD-1 deficiency does not alter ILC-2 cell development. Host CD45.1⁺C57BL6 mice were subjected to total body irradiation (1,050 cGy), and then reconstituted with either CD45.1⁺ WT BM (10 million) or CD45.2⁺Pdcd1^−/− BM (10 million). Some cohorts received both WT and Pdcd1^−/− BM at a 1:1 ratio (5 million WT:5million Pdcd1^−/−). At 8 wk after BM transplant, ILC-2 were analyzed by flow cytometry via gating on Lin⁻ CD45⁺ Thy1⁺ CD127⁺ CD25⁺ ST2⁺ KLRG1⁺ (Representative data from lungs; A–D) and cell numbers were characterized in the lungs, small intestine, and mesenteric LNs. Summary of the absolute numbers of KLRG1⁺ILC-2 cells in the lungs (E), small intestine (F), and mesenteric LNs (G). Single-cell suspensions were stimulated with cytokine stimulation cocktail, and cytokine profile was measured by flow cytometry. Summary of absolute numbers of IL-5 and IL-13 from lung KLRG1⁺ILC-2s (H and I). Host C57BL6 Rag^−/−γc^−/− mice were reconstituted with either CD45.1⁺ WT BM (10 million), or CD45.2⁺Pdcd1^−/− BM (10 million). Some cohorts received both WT and Pdcd1^−/− BM at a 1:1 ratio. At 8 wk after BM transplant, the lung and small intestine were harvested from the mice and KLRG1⁺ILC-2 cell numbers were characterized. Summary of the absolute numbers of KLRG1⁺ILC-2 cells in the lungs (J) and small intestine (K) in the various cohorts was analyzed by flow cytometry. Animals per cohort was n = 5. Data shown are mean ± SEM. Experiments were repeated twice. A one-way ANOVA analysis followed by a multiple comparison test [Turkey] was performed to determine statistical significance between the various cohorts. P ≤ 0.05 was considered significant. Significant p-values are denoted in the figures.