1. Altered distribution of γδT IELs in GPR55-deficient mice.
Altered distribution of γδT IELs in GPR55-deficient mice. (A) Immunofluorescence detection of GL3 (γδTCR), CD8α, laminin, and DAPI with examples of CD8α+GL3+ cells in the PMS and in close association with the epithelium highlighted. Scale bar, 20 μm. (B) Distribution of CD8α+ γδT cells in the small intestine. In the left panels, yellow arrows are used to highlight cells in close association with the epithelium, and orange arrows cells associated with the PMS. Images are representative of five mice of each type. Scale bar, 100 μm. Enlarged images from rectangles in left panels are shown on the right. Scale bar, 25 μm. (C) Percentage of γδ IELs closely associated with epithelium in GPR55 Het and KO mice. (D) Diagram explaining the mixed BM chimera approach used to track control and KO γδT IELs in the same mice. 2P, two-photon; FCM, flow cytometry; IF, immunofluorescence. (E) Immunofluorescence analysis of control (Het, purple arrowheads) and GPR55 KO (red arrowheads) γδT IELs in the small intestine of a mixed BM chimeric mouse (n = 10). Circles show regions where IELs are localized in close association with epithelial cells. Scale bar, 50 μm. (F) Fraction of GPR55 Het and KO γδT IEL, identified as in (E), located in close association with the epithelium. (G) Ratio of γδT cells of KO versus Het origin in the small intestinal IELs and spleen compartments determined by flow cytometry (n = 5). (H) Ratio of γδT cells of KO versus Het origin in the small intestine determined by microscopy as in (E) (n = 10). ***P < 0.001, **P < 0.01, *P < 0.05, n.s. P > 0.05 by Student’s t test (C and F) or one-way ANOVA (G).
Hayakazu Sumida et al. Sci. Immunol. 2017;2:eaao1135
Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works