Figure 5. Inflammatory responses in PAT after skin immunization.

A) Each line in the graphs represents an independent experiment in which the fold-increase in PAT CD11c⁺ inflammatory cells was measured 18 h after epicutaneous application of FITC sensitizer or 18 h after EαGFP was injected i.d. B) Quantitative PCR analysis to determine fold increase in mRNA for inflammatory mediators and key adipose-derived cytokines- TNFα, IL-6, iNOS, TLR4, CCR2, MCP-1, and CD36, 12 h after application of FITC sensitizer to skin. C, D) CD4⁺ T cells were purified from either naïve LNs (+Naïve T) or reactive LNs (+T) of CD45.1⁺ WT mice 4 days after FITC skin painting and transferred into CD45.2⁺ WT mice. These recipients were challenged 5 h later with FITC painting or an irrelevant antigen to which they were naïve (EαGFP) or not challenged (No Antigen). Accumulation of transferred CD45.1⁺ CD4⁺ and endogenous CD45.2⁺ CD4⁺ T cells in the site of skin challenge or the associated PAT was analyzed ∼36 h after challenge. Panel C shows the representative FACS plots of transferred T cells in skin or in PATs. Panel D charts the number of T cells accumulated in PAT of antigen challenged mice, expressed as fold-increase over the number of accumulated in the absence of antigen challenge. **, P < 0.01. All data were obtained from at least 2 independent experiments with more than 3 replicates per group in each experiment.