Figure 6. Transfer of LSIG cells confers immunity to N. brasiliensis infection.

(A-G) Highly purified LSIG cells were transferred into groups of Rag2^−/−Il2rg^−/− hosts. Four weeks after transfer, the indicated groups of mice (n=4-5) were infected with N. brasiliensis and analyzed on day 5 of infection. (A) Total intestinal helminth counts. (B) Percentage (± SD; n=3) of donor-derived (CD45.1⁺) ILC2 in the indicated organs at the day of infection (white bars) and at day 5 after N. brasiliensis infection. (C) Representative dot plot of eosinophil (CD49b^loSiglec-F⁺) accumulation in the lungs of the indicated mouse strains. Numbers in graph represent percentage of cells in the indicated gates. (D) Percentage (± SEM; n=4) of CD49b^loSiglec-F⁺ cells. (E,F) Quantitative RT-PCR analysis (± SD; n=4) of Angiogenin4 (E) and Mucin2 (F) expression by intestinal epithelial cells isolated from the indicated treatment groups. (G) Alcian blue staining of cryosections of the small intestine. Arrowheads indicate goblet cells with high amounts of mucus production. Scale bar, 100 μm. Original magnification, x20.

(H) CCR9 (GFP) expression by intestinal ILC2 and bone marrow ILC2P. Histograms are electronically gated on LSIG cells or ILC2. Open histograms represent analysis of the same population from control mice.

(I,J) Representation of GATA3^hi ILCs (i.e., ILC2 or ILC2P) in C57BL/6 (I, upper panel) and Ccr9-deficient (I, lower panel) mice. Numbers represent percentage of cells in the indicated gates. (J) Percentage (± SEM; n=3) of GATA3^hi ILCs in the indicated organs of control (white bars) or Ccr9-deficient (black bars) mice.

All data are representative of 3 or more independent experiments. * p ≤ 0.05, ** p ≤ 0.01. LSIG: Lin⁻Sca1^hiId2^hiGATA3^hi bone marrow cells.