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. Author manuscript; available in PMC: 2013 Aug 1.
Published in final edited form as: Gastroenterology. 2012 Apr 19;143(2):418–428. doi: 10.1053/j.gastro.2012.04.017

Figure 3. iNKT cells from germ-free animals are hyporesponsive.

Figure 3

(A) Expression of CD69, CD25 and CD5 by iNKT cells from indicated organs derived from germ-free (GF) or specific-pathogen-free (SPF) housed Swiss Webster mice. (B) Expression of CD69 (left panel) and indicated cytokines (right panel) by splenic iNKT cells from GF or SPF housed Swiss Webster mice with or without αGalCer challenge in vivo (90min). The expression of CD69 following αGalCer increased on SPF derived iNKT cells 1.9fold (MFI), whereas the increase on GF derived iNKT cells was lower at 1.75fold (p(SPF +/- αGalCer vs GF +/- αGalCer)= 0.004). (C) Splenocytes from GF and SPF Swiss Webster mice were co-cultured with αGalCer loaded RMA-CD1d cells for 4h and cytokine production by iNKT cells was analyzed by intracellular staining. (D) GF or SPF housed animals on the C57BL/6 background were injected with αGalCer and the cytokine production by splenic iNKT cells was analyzed 90min later. The graph summarizes data from two independent experiments, with 4–5 mice per group. (E) αGalCer-specific in vivo cytotoxicity in spleen 4h after injection of B cell targets into GF or SPF housed Swiss Webster mice. Representative data from two independent experiments are shown. (F) Relative percentage of iNKT cells within TCRβ+ live lymphocytes (left) and of CD127+CD4− iNKT cells (right) from indicated organs of GF or SPF housed Swiss Webster animals. The graphs summarize data from three independent experiments, with 5–8 mice per group.