Extended Data Fig. 10: IL-17 and IL-22 are produced by lung ILCs following Mtb infection and mediate protection through the CXCR5 axis.
From: Group 3 innate lymphoid cells mediate early protective immunity against tuberculosis
C57BL/6 mice were aerosol-infected with approximately 100 CFU Mtb and treated with isotype (n = 5 mice) or anti-NK1.1 (n = 5 mice, PK126, 100 μg) antibodies every 3 days. a, Lung NK cells were determined following treatment with isotype or anti-NK1.1 antibodies at 30 d.p.i. by flow cytometry. b, Lung bacterial burden was assessed at 30 d.p.i. a, b, Significance calculated by Student’s t-test. c, d, Tissues from uninfected Ahrf/f and Ahrf/fRorγtcre mice were collected, and lung and lymph nodes were analysed for the different myeloid (c; alveolar macrophages, myeloid dendritic cells, neutrophils, monocytes and recruited macrophages) and T cell (d; CD3+CD4+, CD3+CD8+, CD3+TCRα+, CD3+γδ+) populations by flow cytometry (n = 9 mice per group). e, Lung cells from C57BL/6 mice were infected in vitro with Mtb (MOI of 0.1) and IL-23 protein levels were measured in supernatants on 5 d.p.i. and compared to uninfected (UI) cells. n = 3 uninfected dishes, n = 4 Mtb-infected dishes. f, Left, lung cells from C57BL/6 mice were infected in vitro with Mtb (MOI of 0.1) as in e and stimulated with recombinant (r)IL-23, rIL-1β, and the protein levels of IL-22 and IL-17 were measured in supernatants and compared with levels in uninfected cells. Right, numbers of IL-17- and IL-22-producing ILCs were measured by flow cytometry. g, C57BL/6 and Cxcr5−/− mice were aerosol-infected with around 100 CFU Mtb and at 30 d.p.i. the bacterial burden was determined in the lungs by plating (n = 5 mice per group). h, ILC3 quantification in FFPE lung sections was carried out by staining with antibodies against CD3, B220 and Rorγt, and the number of Rorγt+CD3− ILC3s were counted and are shown. i, FFPE lung sections from 30 d.p.i. Mtb-infected mice were stained with antibodies against B220 and CD3, and the average size of B cell follicles was quantified. Significance is calculated by Student’s t-test (c–i). Data are mean ± s.d. and individual values. All experiments were replicated at least twice.
