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. Author manuscript; available in PMC: 2019 Aug 1.

Published in final edited form as: J Allergy Clin Immunol. 2017 Nov 9;142(2):647–662. doi: 10.1016/j.jaci.2017.09.042

(A) Expression of IL-17 in CD4 and CD8 T cells (gated on CD3⁺CD4⁺ and CD3⁺CD8⁺ cells) of heat-killed (HK) C. albicans-treated group (right) and PMA-ionomycin-treated group (left) from infected ear skin of wild type B6 mice at day 7 after candida infection. Data are representative of five independent experiments. (B) Expression of IL-17 in CD4 and CD8 T cells (gated on CD3⁺CD4⁺ and CD3⁺CD8⁺ cells) from infected ear skin at day 60 after candida infection when incubated with HK C. albicans for 6 hours in the presence of 50 µg anti-I-A/I-E or 50 µg isotype Abs (rat IgG2b). (C) IL-17 production of γδ and αβ T cells from normal skin prior to candida infection, infected ear skin at day 7 and 30 after candida infection. Data are representative of three independent experiments. (D) Proliferation of CFSE-labeled skin CD4 (middle) and CD8 (right) T cells (gated on CD45.2⁺CD3⁺CD4⁺ and CD45.2⁺CD3⁺CD8⁺ cells) from infected ear skin at day 30 after candida infection. (E) Proliferation of CFSE-labeled LN CD4 (middle) and CD8 (right) T cells (gated on CD45.2⁺CD3⁺CD4⁺ and CD45.2⁺CD3⁺CD8⁺ cells) from draining LN at day 30 after candida infection. CFSE-labeled skin or LN cells were co-cultured 72 hours with CD45.1⁺CD11c⁺ DCs purified from skin draining lymph nodes of day 3 infected mice. Data are representative of three independent experiments. (F) Infected skin samples (n = 4–5) were cleaned with povidone-iodine prior to being harvested at the indicated time after candida infection. C. albicans CFU is expressed as colonies per cm² of infected skin tissues. (G) Normal wild type B6 mice prior to candida infection (naive) and previously infected mice 45 days after candida infection (memory) were re-infected with C. albicans on the ear skin. Half mice were injected daily with FTY720 (1µg/g) during re-infection at day −1, 0, 1, 2. Two days later, skin samples (n = 10 at each group) were harvested and CFU obtained. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant.

(A) Expression of IL-17 in CD4 and CD8 T cells (gated on CD3⁺CD4⁺ and CD3⁺CD8⁺ cells) of heat-killed (HK) C. albicans-treated group (right) and PMA-ionomycin-treated group (left) from infected ear skin of wild type B6 mice at day 7 after candida infection. Data are representative of five independent experiments. (B) Expression of IL-17 in CD4 and CD8 T cells (gated on CD3⁺CD4⁺ and CD3⁺CD8⁺ cells) from infected ear skin at day 60 after candida infection when incubated with HK C. albicans for 6 hours in the presence of 50 µg anti-I-A/I-E or 50 µg isotype Abs (rat IgG2b). (C) IL-17 production of γδ and αβ T cells from normal skin prior to candida infection, infected ear skin at day 7 and 30 after candida infection. Data are representative of three independent experiments. (D) Proliferation of CFSE-labeled skin CD4 (middle) and CD8 (right) T cells (gated on CD45.2⁺CD3⁺CD4⁺ and CD45.2⁺CD3⁺CD8⁺ cells) from infected ear skin at day 30 after candida infection. (E) Proliferation of CFSE-labeled LN CD4 (middle) and CD8 (right) T cells (gated on CD45.2⁺CD3⁺CD4⁺ and CD45.2⁺CD3⁺CD8⁺ cells) from draining LN at day 30 after candida infection. CFSE-labeled skin or LN cells were co-cultured 72 hours with CD45.1⁺CD11c⁺ DCs purified from skin draining lymph nodes of day 3 infected mice. Data are representative of three independent experiments. (F) Infected skin samples (n = 4–5) were cleaned with povidone-iodine prior to being harvested at the indicated time after candida infection. C. albicans CFU is expressed as colonies per cm² of infected skin tissues. (G) Normal wild type B6 mice prior to candida infection (naive) and previously infected mice 45 days after candida infection (memory) were re-infected with C. albicans on the ear skin. Half mice were injected daily with FTY720 (1µg/g) during re-infection at day −1, 0, 1, 2. Two days later, skin samples (n = 10 at each group) were harvested and CFU obtained. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant.

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