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Vaccines development of Vibrio vulnificus. 陳俊良. Immunization with Major Outer Membrane Protein of Vibrio vulnificus Elicits Protective Antibodies in a Murine Model. The Journal of Microbiology, 2005, p.437-442. Cho-Rok Jung, Min-Jung Park and Moon-Soo Heo.
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Immunization with Major Outer Membrane Protein of Vibrio vulnificus Elicits Protective Antibodies in a Murine Model The Journal of Microbiology, 2005, p.437-442. Cho-Rok Jung, Min-Jung Park and Moon-Soo Heo
The presence of capsular polysaccharides (CPS) demonstrated that an ‘antiphagocytic surface antigen’ helps virulent V. vulnificus strains to resist phagocytosis by human polymorphonuclear leukocyte (PMN) Kreger et al., 1981 • During colonization, surface materials of the organisms such as CPS, LPS, and outer membrane proteins (OMP) play an important role in the adhesion to host cells Mirelman, 1986 • Conjugated vaccines between capsular polysaccharide of V. vulnificus (VvPS) - Tetanus toxoid (TT) have been investigated in mouse models Devi et al., 1996 and Morris Jr et al., 1987
OMP (outer membrane protein) • OMP of gram-negative bacteria, which form channels for small hydrophilic molecules, are known as porins • Porins produce water-filled channels across the membrane, such as the OmpF, OmpC, and PhoE of E. coli • It transports of nutrients, antibiotics and other modulators are performed through these channels • OMP possess high hydrophobicity, which is one of important forces that macromolecules use to preserve their binding interaction • OMP can adhere to mucosal membranes and antigen presenting cells of GALT present OMP as antigen to immune cells easily
Motive: • Richardson et al. (1989) identified specific antigens that are expressed in the outer membrane of whole cells or uniquely by in vivo-grown V. cholerae and which react with immune serum and jejunal fluid • More than one cell surface protein antigen of V. cholerae was involved in the induction of protective immunity through the inhibition of intestinal colonization of vibrios • Sciortino (1993) identified 18-kDa cholerae protective antigen which is one of the OMP of V. cholerae • Sperandio et al. (1995) presented evidence that the OmpU of V. cholerae acts as an adherence factor during the colonization of epithelial cells whether MOMP was present as an antigen during the bacterial infection process
The rabbit developed serum antibody reacted with MOMP Coomassie blue staining Western blotting 2, whole bacteria 3, soluble protein 4, insoluble protein 1, whole bacteria 2, soluble protein 3, insoluble protein MOMP, major outer membrane protein
Molecular size of MOMP was identified as 36-kDa on 13% SDS-PAGE
N-terminal sequences of MOMP revealed a high degreesimilarity with OmpU of V. vulnificus CMCP6 and YJ016
Immunization with MOMP produced specific IgG and IgM IgG IgM MOMP stimulated humoral immune response
MOMP may have stimulated the complementsystem in cooperation with Alum
protective immunity of MOMP against V. vulnificus infection BALB/c mice (n = 10) were immunized with 40 μg of MOMP. infected with V. vulnificus (107) after immunization (IP) immunized mice were healthy until 28 days after the inoculation
Discussion: • Most bacterial porins were expected to be strong immunogens that stimulated the host immune system. • Galdiero et al., reported that porins of Salmonella typhimurium stimulated Th1 and Th2 cells and IL-1 and IL-6 gene expression in mice. • Gordon and Alistair reported OMP like porins might be a candidate vaccine due to the hydrophobicity and immunogenicity of this molecule.