We examined whether two types of xylooligosaccharides (neutral or acidic xylooligosaccharides) derived from hardwood kraft pulp ameliorate the development of atopic dermatitis (AD)-like skin lesions induced by repeated application of picryl chloride (PiCl) in NC/Nga mice. Oral administration of acidic xylooligosaccharides at a daily dose of 100 mg/kg significantly prevented the development of AD-like skin lesions. Serum histamine level was significantly suppressed, but serum total IgE level was not significantly suppressed. Moreover, the secretion of inflammatory cytokine IL-12 from splenic lymphocytes was significantly suppressed. On the other hand, neutral xylooligosaccharides showed no significant preventive effect on the development of AD-like symptoms. These results suggest that oral administration of acidic xylooligosaccharides may be effective in preventing the development of AD-like skin disease and one of the mechanisms is the suppressive effect on IL-12.