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Human neutrophil peptide (HNP) defensins were studied to determine their potential effects on adaptive mucosal immunity. Intranasal delivery of HNPs plus ovalbumin (OVA) enhanced OVA-specific serum IgG antibody (Ab) responses. However, OVA-specific IgA Abs were not induced in mucosal secretions or in serum. CD4⁺ T cells of intranasally immunized mice displayed higher OVA-specific proliferative responses and elevated production of interferon γ, interleukin (IL) 5, IL-6, and IL-10 when compared with control groups receiving OVA alone. In vitro, HNPs also enhanced both proliferative responses and T helper (Th) cytokine secretion profiles of CD3ɛ-stimulated spleen- and Peyer’s patch-derived naive CD4⁺ T cells. HNPs modulated the expression of costimulatory molecules by lipopolysaccharide- or CD3ɛ-stimulated splenic and Peyer’s patch B or T cell populations, respectively. These studies show that …