IL-16 is a natural ligand of CD4 molecules and induces chemotaxis in CD4-expressing cells. It amplifies the inflammatory reaction by stimulating cytokine production in monocytes and activating T-cells. There is evidence that IL-16 plays a role in the pathogenesis of atopic dermatitis, and increased serum levels of IL-16 have been detected in allergic diseases. However, few data are available on IL-16 serum levels in atopic dermatitis. The aim of our study is to measure IL-16 serum levels in childhood atopic dermatitis before and after treatment and to evaluate a possible correlation between IL-16 serum levels and disease severity. IL-16 serum levels were measured by an ELISA approach in 34 children (19 males and 15 females; mean age 6.8 years) with moderate to severe atopic dermatitis, at their first visit and after 3 months of treatment, and in 10 non-atopic healthy controls of the same age group. The severity of atopic dermatitis was measured by SCORAD index. IL-16 serum levels were significantly higher in patients affected by atopic dermatitis than in controls before and after treatment with tacrolimus ointment. No clear correlation was found between IL-16 serum levels and atopic dermatitis severity. IL-16 serum levels are increased in atopic dermatitis but do not seem to correlate with disease severity.