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. Author manuscript; available in PMC: 2023 Feb 23.
Published in final edited form as: Contact Dermatitis. 2022 Mar 18;87(1):28–39. doi: 10.1111/cod.14088

FIGURE 1.

FIGURE 1

Enhanced CHS in obese C57BL/6 mice. CHS was induced after 8 weeks of feeding with HFD or ND. (A) Weight gain in mice fed with ND (group A) and HFD (group B); n = 12/group. (B) Ear swelling and ear weight in mice fed with ND (group A and B) or HFD (group C and D). Mice were sensitized (day 0) by application of 5% TNCB to the shaved abdomen (group B and D). Unsensitized mice fed with ND (group A) or HFD (group C) were used as negative controls. CHS and ear weight were measured 24 hours after challenge with TNCB (day +4); n = 12/group. (C) Histology of ear tissue after challenge with hapten TNCB in unsensitized mice on the ND (group A) and HFD (group C) and in sensitized mice fed with ND (group B) and HFD (group D). Hematoxylin and eosin staining. Scale bar = 50 μm. Representative images (magnification: ×40). (D) Concentrations of IFN-γ, IL-17A, and TNF-α in ELNC culture supernatants. Mice were fed with ND (group B) or HFD (group D) and subsequently sensitized and challenged with hapten TNCB. Negative control mice were fed with ND (Group A) or HFD (Group C) and subsequently challenged with hapten. ELNCs were isolated 24 hours after challenge and were cultured with antigen for 48 hours. The culture supernatants of ELNCs were collected and tested for cytokine concentration using an ELISA Kit; n = 5–7/group. (E), MPO activity in ear tissue after challenge with hapten TNCB in unsensitized mice (negative) on the ND (group A) and HFD (group C) and in TNCB-sensitized mice (positive) fed with ND (group B) and HFD (group D). MPO colorimetric activity assay was used to determine the activity of MPO in samples; n = 6–12/group. (F,G) Concentration of anti-TNP IgG1 and IgG2c antibodies in serum after challenge with hapten TNCB in unsensitized mice (negative) on the ND (group A) and HFD (group C) and in TNCB-sensitized (positive) mice fed with ND (group B) and HFD (group D). The collected sera were tested for antibodies concentration using an ELISA; n = 6–12/group. *P < .05, **P < .01, ***P < .001. Abbreviations: CHS, contact hypersensitivity reaction; ELNC, auricular lymph node cells; HFD, high fat diet; MPO, myeloperoxidase; ND, normal diet; TNCB, 2,4,6-trinitrochlorobenzene; TNP, 2,4,6-trinitrophenyl