ABSTRACT
To provide basic information on the pathogengenesis of Vibrio vulnificus infection and for the manufacture of effective vaccine, outer membrane proteins (OMPs) were extracted from V. vulnificus ATCC 27562 strain and analysed by 12% of sodium dodecyl sulfate-polyarylamide gel(SDS-PAGE). Major 36 kDa and 48 kDa, 46 kDa, 66 kDa and 24 kDa protein bands appeared on gel by Coomassie stain and determined by densitometer. Immunogenecity of these proteins was examined by enzyme-linked immunosorbant assay(ELISA) and western blotting with rabbit anti-V. vulnificus serum. OMPs reacted with this antiserum, and the major 36 kDa protein appeared most immunogenic.
Subject(s)
Blotting, Western , Enzyme-Linked Immunosorbent Assay , Membrane Proteins , Membranes , Sodium , Vibrio vulnificusABSTRACT
BACKGROUND: We developed a Pseudomonas aeruginosa outer membrane protein(OMP) vaccine, CFC-101, and the prophylactic efficacy of which has been demonstrated in animal models. In order to evaluate the safety and immunogenicity of the P. aeruginosa vaccine, we carried out a phase I/IIa clinical trial in healthy male volunteers. METHODS: Groups of eight volunteers, including two placebo subjects, were vaccinated intramuscularly with three doses of 0.25, 0.5 or 1.0 mg of the vaccine at one week intervals. Signs of systemic and local reactions observed after vaccination were recorded for each vaccinee for 5 days. Physical examinations were performed on days 0, 1, 7, 8, 14, 15, 21, and 42, and clinical laboratory tests were done on days 0, 3, and 21. Blood samples for assay of serum antibody levels were obtained up to 42 days after the first vaccination. RESULTS: The vaccine was generally well tolerated by all vaccinees, showing no significant side effects. In the three dosage groups, all vaccinees, except one receiving the 0.25 mg dose, showed significant elevation in serum IgG antibody titers against the vaccine proteins, indicating 100% seroconversion in 0.5 and 1.0 mg groups. The human antibodies induced by the vaccine were specific for P. aeruginosa OMPs, as confirmed by western blot analysis and immunoprecipitation assays. The capacity of the human antisera to enhance opsonophagocytic killing activity by polymorphonuclear leukocytes and to confer protection against P. aeruginosa infections indicates that the antibodies elicited by the vaccine have protective efficacy. CONCLUSION: We conclude that the P. aeruginosa OMP vaccine is safe and effective for human use and its optimal dose to be 0.5 or 1.0 mg.
Subject(s)
Humans , Male , Antibodies , Blotting, Western , Homicide , Immune Sera , Immunoglobulin G , Immunoprecipitation , Membrane Proteins , Membranes , Models, Animal , Neutrophils , Physical Examination , Pseudomonas aeruginosa , Pseudomonas , Vaccination , VolunteersABSTRACT
Shigella sonnei KNIH104S, which was selected by Korean National Institute of Health, expresses form I-antigen as a somatic antigen. In this study, we cloned the genes responsible for form I-antigen synthesis from S. sonnei KNIH104S. A Sau3AI-generated cosmid library of S. sonnei KNIH104S plasmids were transfected into E. coli LE392 and transfectants were tested for agglutination with antiserum against S. sonnei form I-antigen. A clone, JH222, showing the strongest agglutination activity was chosen for further analysis. A recombinant cosmid, pJH222, was isolated from the strain JH222 and retransfected into E. coli LE392. All of the transfectants agglutinated with antiserum against form I-antigen, indicating that pJH222 carried the genes required for S. sonnei form I-antigen synthesis. Restriction analysis of pJH222 revealed a 38 kb insert, which was confirmed by Southern hybridization analysis to be present on a large plasmid of S. sonnei KNIH104S.