Intranasal Administration of Recombinant Neisseria gonorrhoeae Transferrin Binding Proteins A and B Conjugated to the Cholera Toxin B Subunit Induces Systemic and Vaginal Antibodies in Mice

Department of Microbiology and Immunology, Medical College of Virginia Campus of Virginia Commonwealth University, Richmond, Virginia 23298-0678, USA.
Infection and Immunity (Impact Factor: 3.73). 08/2005; 73(7):3945-53. DOI: 10.1128/IAI.73.7.3945-3953.2005
Source: PubMed


The transferrin binding proteins (TbpA and TbpB) comprise the gonococcal transferrin receptor and are considered potential antigens for inclusion in a vaccine against Neisseria gonorrhoeae. Intranasal (IN) immunization has shown promise in development of immunity against sexually transmitted disease pathogens, in part due to the induction of antigen-specific genital tract immunoglobulin A (IgA) and IgG. Conjugation of antigens to the highly immunogenic cholera toxin B subunit (Ctb) enhances antibody responses in the serum and mucosal secretions following IN vaccination. In the current study, we characterized the anti-Tbp immune responses following immunization of mice IN with recombinant transferrin binding proteins (rTbpA and rTbpB) conjugated to rCtb. We found that both rTbpA-Ctb and rTbpB-Ctb conjugates administered IN induced antibody responses in the serum and genital tract. IN immunization resulted in both IgA and IgG in the genital tract; however, subcutaneous immunization mainly generated IgG. Surprisingly, rTbpA alone was immunogenic and induced serum and mucosal antibody responses similar to those elicited against the rTbpA-Ctb conjugate. Overall, rTbpB was much more immunogenic than rTbpA, generating serum IgG levels that were greater than those elicited against rTbpA. Bactericidal assays conducted with sera collected from mice immunized IN with TbpA and/or TbpB indicated that both antigens generated antibodies with bactericidal activity. Anti-TbpA antibodies were cross-bactericidal against heterologous gonococcal strains, whereas TbpB-specific antibodies were less cross-reactive. By contrast, antibodies elicited via subcutaneous immunization were not cross-bactericidal against heterologous strains, indicating that IN vaccination could be the preferred route for elicitation of biologically functional antibodies.

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    • "A variety of surface antigens, e.g., pilus [8]–[12], lipooligosaccharide (LOS) [13]–[15], opacity-associated protein (Opa) [16], porins [17]–[19], transferrin-binding proteins [20]–[21], surface protein A (NspA) [22]–[23], lipoproteins [24], outer membrane preparations [25], have been used to develop vaccines for N. gonorrhoeae. Two of these antigens have been in clinical trials and few have been tested in animal models. "
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    ABSTRACT: Neisseria gonorrhoeae (N. gonorrhoeae) outer membrane protein reduction modifiable protein (Rmp) has strong immunogenicity. However, anti-Rmp antibodies block rather than preserve the antibacterial effects of protective antibodies, which hampers the development of vaccines for gonococcal infections. We herein constructed an Rmp deletion mutant strain of N. gonorrhoeae by gene homologous recombination. The 261-460 nucleotide residues of Rmp gene amplified from N. gonorrhoeae WHO-A strain were replaced with a kanamycin-resistant Kan gene amplified from pET-28a. The resultant hybridized DNA was transformed into N. gonorrhoeae WHO-A strain. PCR was used to screen the colonies in which wild-type Rmp gene was replaced with a mutant gene fragment. Western blotting revealed that the Rmp deletion mutant strain did not express Rmp protein. Rmp deletion did not alter the morphological and Gram staining properties of the mutant strain that grew slightly more slowly than the wild-type one. Rmp gene mutated stably throughout 25 generations of passage. Antibody-mediated complement-dependent cytotoxicity assay indicated that the antibodies induced by the mutant strain had evidently higher bactericidal activities than those induced by the wild-type strain. Further modification of the Rmp deletion mutant strain is still required in the development of novel live attenuated vaccines for gonorrhea by Opa genes deletion or screening of phenotypic variant strains that do not express Opa proteins.
    PLoS ONE 03/2014; 9(3):e90525. DOI:10.1371/journal.pone.0090525 · 3.23 Impact Factor
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    • "Antigenic and sequence variability of TbpB proteins is more extensive (Cornelissen et al., 1997a), but neither protein is subject to high-frequency phase or antigenic variation, as is the case with many other neisserial surface antigens . Both transferrin binding proteins are antigenic when animals are immunized with the purified proteins (Price et al., 2005; 2007); however, natural gonococcal infections , which do not elicit protective immunity, also do not generate high titre anti-Tbp antibodies (Price et al., 2004). Meningococcal transferrin binding proteins are immunogenic in both animals (Rokbi et al., 1997) and humans (Gorringe et al., 1995), which is consistent with the hypothesis that gonococci are capable of immune suppression during infection (Liu et al., 2011). "
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    ABSTRACT: Two pathogenic species within the genus Neisseria cause the diseases gonorrhoea and meningitis. While vaccines are available to protect against four N. meningitidis serogroups, there is currently no commercial vaccine to protect against serogroup B or against N. gonorrhoeae. Moreover, the available vaccines have significant limitations and with antibiotic resistance becoming an alarming issue, the search for effective vaccine targets to elicit long-lasting protection against Neisseria species is becoming more urgent. One strategy for vaccine development has targeted the neisserial iron import systems. Without iron, the Neisseriae cannot survive and, therefore, these iron import systems tend to be relatively well conserved and are promising vaccine targets, having the potential to offer broad protection against both gonococcal and meningococcal infections. These efforts have been boosted by recent reports of the crystal structures of the neisserial receptor proteins TbpA and TbpB, each solved in complex with human transferrin, an iron binding protein normally responsible for delivering iron to human cells. Here, we review the recent structural reports and put them into perspective with available functional studies in order to derive the mechanism(s) for how the pathogenic Neisseriae are able to hijack human iron transport systems for their own survival and pathogenesis.
    Molecular Microbiology 09/2012; 86(2):246-57. DOI:10.1111/mmi.12002 · 4.42 Impact Factor
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    • "One way to do this is being explored by one of us (Ann E. Jerse), utilizing circular loop peptides as the immmunogens, based on evidence that analagous circular Opa peptides are able to elicit potentially protective immune responses (Cole and Jerse, 2009). Intranasal immunization with the GC transferrin-binding proteins TbpA or TbpB, or both, elicited bactericidal immune responses; TbpA stimulated more broadly cross-reactive antibodies than did TbpB (Price et al., 2005). Immunization of mice with genetic chimeras that fused parts of TbpA and TbpB stimulated production of vaginal antibodies that inhibited growth in vitro (Price et al., 2007). "
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    ABSTRACT: Immune responses to the gonococcus after natural infection ordinarily result in little immunity to reinfection, due to antigenic variation of the gonococcus, and redirection or suppression of immune responses. Brinton and colleagues demonstrated that parenteral immunization of male human volunteers with a purified pilus vaccine gave partial protection against infection by the homologous strain. However, the vaccine failed in a clinical trial. Recent vaccine development efforts have focused on the female mouse model of genital gonococcal infection. Here we discuss the state of the field, including our unpublished data regarding efficacy in the mouse model of either viral replicon particle (VRP) vaccines, or outer membrane vesicle (OMV) vaccines. The OMV vaccines failed, despite excellent serum and mucosal antibody responses. Protection after a regimen consisting of a PorB-VRP prime plus recombinant PorB boost was correlated with apparent Th1, but not with antibody, responses. Protection probably was due to powerful adjuvant effects of the VRP vector. New tools including novel transgenic mice expressing human genes required for gonococcal infection should enable future research. Surrogates for immunity are needed. Increasing antimicrobial resistance trends among gonococci makes development of a vaccine more urgent.
    Frontiers in Microbiology 06/2011; 2:124. DOI:10.3389/fmicb.2011.00124 · 3.99 Impact Factor
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