Intranasal administration of proteoliposome-derived cochleates from Vibrio cholerae O1 induce mucosal and systemic immune responses in mice

• Published in: Methods (San Diego, Calif.) Calif.), 2009-12, Vol.49 (4), p.309-315
• Main Authors: Acevedo, Reinaldo, Callicó, Adriana, del Campo, Judith, González, Elizabeth, Cedré, Bárbara, González, Lissette, Romeu, Belkis, Zayas, Caridad, Lastre, Miriam, Fernández, Sonsire, Oliva, Reynaldo, García, Luis, Pérez, José Luis, Pérez, Oliver
• Format: Article
• Language: English
• Subjects: Adjuvant; Administration, Intranasal; Animals; Cochleates; Deoxycholic Acid - administration & dosage; Deoxycholic Acid - immunology; Edetic Acid - administration & dosage; Edetic Acid - immunology; Female; Immunity, Mucosal - drug effects; Immunity, Mucosal - immunology; Immunoglobulin G - immunology; Intranasal; Mice; Mice, Inbred BALB C; Mucosal; Mucous Membrane - drug effects; Mucous Membrane - immunology; Proteolipids - administration & dosage; Proteolipids - immunology; Proteoliposomes; Vaccine; Vesicle; Vibrio cholerae; Vibrio cholerae O1 - immunology
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1016/j.ymeth.2009.03.027

Conservative estimates place the death toll from cholera at more than 100,000 persons each year. A particulate mucosal vaccine strategy combining antigens and immune stimulator molecules from Vibrio cholerae to overcome this problem is described. Proteoliposomes extracted from V. cholerae O1 were transformed into cochleates (AFCo2, Adjuvant Finlay cochleate 2) through a calcium inducible rotary dialysis method. Light microscopy was carried out and tubules of 16.25 ± 4.57 μm in length were observed. Western blots were performed to verify the immunochemical properties of the main AFCo2 incorporated antigens, revealing full recognition of the outer membrane protein U (OmpU), lipopolysaccharide (LPS), and mannose-sensitive hemagglutinin (MSHA) antigens. AFCo2 were administered by the intranasal route using a two or three dose schedule and the immune response against V. cholerae antigens was assessed. Three AFCo2 doses were required to induce significant ( p < 0.05), antigen specific IgA in saliva (1.34 ± 0.135) and feces (0.60 ± 0.089). While, two or three doses of AFCo2 or proteoliposomes induce similar specific IgG and vibriocidal activity responses in sera. These results show for the first time that AFCo2 can be obtained from V. cholerae O1 proteoliposomes and have the potential to protect against the pathogen when administered intranasally.