Induction of mucosal immune response after intranasal or oral inoculation of mice with Lactococcus lactis producing bovine beta-lactoglobulin

The bovine beta-lactoglobulin (BLG) is a major cow's milk allergen. Here, w e evaluated the immune response against BLG induced in mice, using the orga nism Lactococcus lactis, which has GRAS ("generally regarded as safe") stat us, as a delivery vehicle, The cDNA of the big gene, encoding BLG, was expr essed and engineered for either intra- or extracellular expression in L. la ctis, Using a constitutive promoter, the yield of intracellular recombinant BLG (rBLG) was about 20 ng per mi of culture. To increase the quantity of rBLG, the nisin-inducible expression system was used to produce rBLG in the cytoplasmic and extracellular locations. Although the majority of rBLG rem ained in the cytoplasm, the highest yield (2 mug per mi of culture) was obt ained with a secreting strain that encodes a fusion between a lactococcal s ignal peptide and rBLG, Whatever the expression system, the rBLG is produce d mostly in a soluble, intracellular, and denatured form. The BLG-producing strains were then administered either orally or intranasally to mice, and the immune response to BLG was examined. Specific anti-BLG immunoglobulin A (IgA) antibodies were detected 3 weeks after the immunization protocol in the feces of mice immunized with the secreting lactococcal strain. Specific anti-BLG IgA detected in mice immunized with lactococci was higher than th at obtained in mice immunized with the same quantity of pure BLG. No specif ic anti-BLG IgE, IgA, IgG1, or IgG2a was detected in sera of mice, These re combinant lactococcal strains constitute good vehicles to induce a mucosal immune response to a model allergen and to better understand the mechanism of allergy induced by BLG.