Validation of the protective effects of nasal immunisation against NHBA pro-tein of Neisseria gonorrhoeae
Objective To express and purify Neisseria gonorrhoeae NHBA protein,and to prepare polyclonal an-tibodies against NHBA by the nasal route of immunization and evaluate the immuno-protective effects of NHBA antisera.Methods Constructed prokaryotic expression strains of NHBA protein and carried out the expression and purification of target proteins.Mice were randomized into NHBA group:NHBA protein+cholera toxin B subunit(CTB)adjuvant;phosphate buffer solution(PBS)group:PBS+CTB adjuvant.Polyclonal antibodies against NHBA protein were produced by intranasal immunization of mice and ELISA was used to determine IgG antibody potency in NHBA antisera.NHBA protein expression was verified in Neisseria gonorrhoeae strains by Western Blot.The immune-protective effect of NHBA antisera was assessed using serum bactericidal activity assay,Op-sonophagocytic activity assay and antibody adhesion inhibition assay.Results NHBA protein was successfully pro-duced and purified.High titers of IgG antibody were produced in the serum after nasal immunization of mice.Western Blot showed that the antiserum recognized the naturally expressed NHBA protein in the Neisseria gonor-rhoeae strain.The effective bactericidal titers in the serum bactericidal assay and the opsonophagocytic activity as-say were 1/200 and 1/400,respectively.The result of the antibody adhesion inhibition assay showed that the NH-BA antiserum effectively inhibited the adhesion of Neisseria gonorrhoeae to ME-180 cells.Conclusion Neisseria gonorrhoeae NHBA protein immunized nasally to obtain antisera against NHBA,NHBA antisera showed immuno-protective effects,and NHBA could be used as effective targets for Neisseria gonorrhoeae nasal mucosal vaccines in the future.
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