title: Development of a cost-effective vaccine candidate with outer membrane vesicles of a tolA-disrupted Shigella boydii strain.
abstract: Our previous studies on outer membrane vesicles based vaccine development against shigellosis, revealed the inability of Shigella to release significant amount of vesicles naturally, during growth. Disruption of tolA, one of the genes of the Tol-Pal system of Gram negative bacterial membrane, has increased the vesicle release rate of a Shigella boydii type 4 strain to approximately 60% higher. We also noticed the vesicles, released from tolA-disrupted strain captured more OmpA protein and lipopolysaccharide, compared to the vesicles released from its wild type prototype. Six to seven weeks old BALB/c mice, immunized with 25 μg of three oral doses of the vesicles, released by tolA mutant, conferred 100% protection against lethal homologous challenge through nasal route, compared to only 60% protection after the same dose of wild type immunogen. Mice, immunized with the vesicles from tolA-mutant, manifested significant secretion of mucosal IgG and IgA. A sharp and significant response of pro-inflammatory cytokines (TNF-α, IL-6, IFN-γ) were also observed in the lung lavage of these groups of mice, within 6h post challenge; but at 24h, these inflammatory cytokines showed the sign of subsidence and the system was taken over by the release of anti-inflammatory cytokines (IL-4 and IL-10). Studies with naïve peritoneal macrophages, proved further, the potency of these vesicles to stimulate nitric oxide and TNF-α, IL-12p70, IL-6 and IL-10 productions in-vitro. The ability of these vesicles to trigger polarization of CD4(+) T cells toward Th1 adaptive immune response, had also been observed along with the presence of anti-inflammatory cytokines in the system. Our study demonstrated, the vesicles from tolA-disrupted Shigella were able to suppress Shigella-mediated inflammation in the host and could balance between inflammation and anti-inflammation, promoting better survival and health of the infected mice. Outer membrane vesicles from tolA-mutant, could be a potential cost-effective vaccine candidate against shigellosis.
authors.affiliation: Division of Bacteriology I, National Institute of Infectious Diseases, Tokyo, Japan; Division of Bacteriology, National Institute of Cholera and Enteric Diseases, Kolkata, India.
authors.forename: Soma
authors.initials: S
authors.lastname: Mitra
authors.affiliation: Division of Bacteriology, National Institute of Cholera and Enteric Diseases, Kolkata, India.
authors.forename: Ritam
authors.initials: R
authors.lastname: Sinha
authors.affiliation: Division of Bacteriology I, National Institute of Infectious Diseases, Tokyo, Japan.
authors.forename: Jiro
authors.initials: J
authors.lastname: Mitobe
authors.affiliation: Division of Bacteriology, National Institute of Cholera and Enteric Diseases, Kolkata, India. Electronic address: hemantakoley@hotmail.com.
authors.forename: Hemanta
authors.initials: H
authors.lastname: Koley
mesh_terms: D000284:Administration, Oral; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000737:chemistry; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D016207:Cytokines / Q000737:chemistry; D004405:Dysentery, Bacillary / Q000517:prevention & control; D000067128:Extracellular Vesicles / Q000276:immunology*; D055786:Gene Knockout Techniques; D008168:Lung / Q000276:immunology; D017737:Macrophages, Peritoneal / Q000276:immunology; D008807:Mice, Inbred BALB C; D022281:Shigella Vaccines / Q000276:immunology*; D012761:Shigella boydii / Q000235:genetics
chemical_list: D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D016207:Cytokines; D022281:Shigella Vaccines; C078167:OMPA outer membrane proteins
keywords: Cytokines; Lipopolysaccharide; Outer membrane protein A; Outer membrane vesicles; Protective efficacy; Shigella; tolA