Active and Passive Intranasal Immunizations with Streptococcal Surface Protein C5a Peptidase Prevent Infection of Murine Nasal Mucosa-Associated Lymphoid Tissue, a Functional Homologue of Human Tonsils

ABSTRACT C5a peptidase, also called SCPA (surface-bound C5a peptidase), is a surface-bound protein on group A streptococci (GAS), etiologic agents for a variety of human diseases including pharyngitis, impetigo, toxic shock, and necrotizing fasciitis, as well as the postinfection sequelae rheumatic fever and rheumatic heart disease. This protein is highly conserved among different serotypes and is also expressed in human isolates of group B, C, and G streptococci. Human tonsils are the primary reservoirs for GAS, maintaining endemic disease across the globe. We recently reported that GAS preferentially target nasal mucosa-associated lymphoid tissue (NALT) in mice, a tissue functionally analogous to human tonsils. Experiments using a C5a peptidase loss-of-function mutant and an intranasal infection model showed that this protease is required for efficient colonization of NALT. An effective vaccine should prevent infection of this secondary lymphoid tissue; therefore, the potential of anti-SCPA antibodies to protect against streptococcal infection of NALT was investigated. Experiments showed that GAS colonization of NALT was significantly reduced following intranasal immunization of mice with recombinant SCPA protein administered alone or with cholera toxin, whereas a high degree of GAS colonization of NALT was observed in control mice immunized with phosphate-buffered saline only. Moreover, administration of anti-SCPA serum by the intranasal route protected mice against streptococcal infection. These results suggest that intranasal immunization with SCPA would prevent colonization and infection of human tonsils, thereby eliminating potential reservoirs that maintain endemic disease.

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Protection from Group B Streptococcal Infection in Neonatal Mice by Maternal Immunization with Recombinant Sip Protein

Infection and Immunity ◽

10.1128/iai.70.9.4897-4901.2002 ◽

2002 ◽

Vol 70 (9) ◽

pp. 4897-4901 ◽

Cited By ~ 42

Author(s):

Denis Martin ◽

Stéphane Rioux ◽

Edith Gagnon ◽

Martine Boyer ◽

Josée Hamel ◽

...

Keyword(s):

Protective Immunity ◽

Group B Streptococcus ◽

Surface Protein ◽

Neonatal Infection ◽

Active Immunization ◽

Infection Model ◽

Lethal Challenge ◽

Specific Antibodies ◽

Pregnant Mice ◽

Group B

ABSTRACT The protective potential of antibodies directed against group B streptococcus (GBS) Sip surface protein was determined by using the mouse neonatal infection model. Rabbit Sip-specific antibodies administered passively to pregnant mice protected their pups against a GBS lethal challenge. In addition, active immunization with purified recombinant Sip protein of female CD-1 mice induced the production of specific antibodies that also confer protection to the newborn pups against GBS strains of serotypes Ia/c, Ib, II, III, and V. These data confirm that Sip-specific antibodies can cross the placenta and conferred protective immunity against GBS infections.

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In Vivo Activation of Naive CD4+ T Cells in Nasal Mucosa-Associated Lymphoid Tissue following Intranasal Immunization with Recombinant Streptococcus gordonii

Infection and Immunity ◽

10.1128/iai.74.5.2760-2766.2006 ◽

2006 ◽

Vol 74 (5) ◽

pp. 2760-2766 ◽

Cited By ~ 23

Author(s):

Donata Medaglini ◽

Annalisa Ciabattini ◽

Anna Maria Cuppone ◽

Caterina Costa ◽

Susanna Ricci ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Lymph Nodes ◽

Nasal Mucosa ◽

T Cell Activation ◽

Lymphoid Tissue ◽

Streptococcus Gordonii ◽

Cervical Lymph Nodes ◽

Intranasal Immunization

ABSTRACT The antigen-specific primary activation of CD4+ T cells was studied in vivo by adoptive transfer of ovalbumin-specific transgenic T cells (KJ1-26+ CD4+) following intranasal immunization with recombinant Streptococcus gordonii. A strain of S. gordonii expressing on its surface a model vaccine antigen fused to the ovalbumin (OVA) peptide from position 323 to 339 was constructed and used to study the OVA-specific T-cell activation in nasal mucosa-associated lymphoid tissue (NALT), lymph nodes, and spleens of mice immunized by the intranasal route. The recombinant strain, but not the wild type, activated the OVA-specific CD4+ T-cell population in the NALT (89% of KJ1-26+ CD4+ T cells) just 3 days following immunization. In the cervical lymph nodes and in the spleen, the percentage of proliferating cells was initially low, but it reached the peak of activation at day 5 (90%). This antigen-specific clonal expansion of KJ1-26+ CD4+ T cells after intranasal immunization was obtained with live and inactivated recombinant bacteria, and it indicates that the NALT is the site of antigen-specific T-cell priming.

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Intranasal Immunization of Mice with Group B Streptococcal Protein Rib and Cholera Toxin B Subunit Confers Protection against Lethal Infection

Infection and Immunity ◽

10.1128/iai.72.2.1184-1187.2004 ◽

2004 ◽

Vol 72 (2) ◽

pp. 1184-1187 ◽

Cited By ~ 21

Author(s):

Charlotte Larsson ◽

Jan Holmgren ◽

Gunnar Lindahl ◽

Charlotta Bergquist

Keyword(s):

Cholera Toxin ◽

Group B Streptococcus ◽

Surface Protein ◽

Cholera Toxin B Subunit ◽

Toxin B ◽

Intranasal Immunization ◽

Cholera Toxin B ◽

B Subunit ◽

A Cell ◽

Group B

ABSTRACT Intranasal immunization of mice with Rib, a cell surface protein of group B streptococcus (GBS), conjugated to or simply coadministered with the recombinant cholera toxin B subunit, induces systemic immunoglobulin G (IgG) and local IgA antibody responses and confers protection against lethal GBS infection. These findings have implications for the development of a human GBS vaccine.

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Intranasal Immunization with Cytotoxic T-Lymphocyte Epitope Peptide and Mucosal Adjuvant Cholera Toxin: Selective Augmentation of Peptide-Presenting Dendritic Cells in Nasal Mucosa-Associated Lymphoid Tissue

Infection and Immunity ◽

10.1128/iai.66.12.5876-5881.1998 ◽

1998 ◽

Vol 66 (12) ◽

pp. 5876-5881 ◽

Cited By ~ 50

Author(s):

Angel Porgador ◽

Herman F. Staats ◽

Yasushi Itoh ◽

Brian L. Kelsall

Keyword(s):

Dendritic Cells ◽

Cholera Toxin ◽

Nasal Mucosa ◽

Lymphoid Tissue ◽

Cytotoxic T Lymphocyte ◽

Epitope Peptide ◽

Cervical Lymph Nodes ◽

Mucosal Adjuvant ◽

Intranasal Immunization ◽

Ctl Epitope

ABSTRACT We previously reported that cholera toxin (CT) was required as a mucosal adjuvant for the induction of peptide-specific cytotoxic T lymphocytes (CTL) following intranasal immunization with CTL epitope peptides (A. Porgador et al., J. Immunol. 158:834–841, 1997). The present study was performed to identify the site and the antigen-presenting cell (APC) population responsible for the presentation of intranasally administered CTL epitope peptide immunogens and to determine whether CT directly affects antigen presentation by these APCs. For these experiments, C57BL/6 mice were intranasally immunized with the ovalbumin H-2Kb-restricted CTL epitope SIINFEKL with or without CT. Cells were then isolated from the cervical lymph nodes (CLN) and the nasal mucosa-associated lymphoid tissue (NALT) and tested for the ability to stimulate the B3Z T-cell hybridoma, which recognizes SIINFEKL in association with H-2Kb. Dendritic cell (DC)-enriched CLN cells from mice immunized with peptide and CT or peptide only could stimulate B3Z cells, while DC-depleted CLN cells from either group were unable to stimulate B3Z cells. NALT cells of mice immunized with peptide and CT, but not with peptide alone, were able to efficiently stimulate B3Z hybridomas. Depletion of N418-positive DC from these NALT cells resulted in significant reduction of B3Z activation. Our results indicate that DC are the APC responsible for the presentation of CTL epitope peptides following intranasal immunization and that CT augments the ability of dendritic cells in the NALT, but not in the draining CLN, to present CLT epitope peptides. This finding suggests that CT acts locally as a mucosal adjuvant and that NALT DC are the predominant APC involved with the induction of immunity after intranasal immunization with peptide immunogens and CT.

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Surface Protein EF3314 Contributes to Virulence Properties of Enterococcus faecalis

The International Journal of Artificial Organs ◽

10.1177/039139880903200910 ◽

2009 ◽

Vol 32 (9) ◽

pp. 611-620 ◽

Cited By ~ 12

Author(s):

Roberta Creti ◽

Francesca Fabretti ◽

Stefanie Koch ◽

Johannes Huebner ◽

Danielle A. Garsin ◽

...

Keyword(s):

Amino Acids ◽

Enterococcus Faecalis ◽

Polyclonal Antibodies ◽

Gc Content ◽

Surface Protein ◽

Animal Origin ◽

Infection Model ◽

Group B Streptococci ◽

Group B ◽

Virulence Properties

Purpose Identification of putative new virulence factors as additional targets for therapeutic approaches alternative to antibiotic treatment of multi-resistant enterococcal infections. Methods The EF3314 gene, coding for a putative surface-exposed antigen, was identified by the analysis of the Enterococcus faecalis V583 genome for LPXTG-motif cell wall anchor surface protein genes. A non-polar EF3314 gene deletion mutant in the E. faecalis 12030 human clinical isolate was obtained. The wild type and the isogenic mutant strain were investigated for biofilm formation, adherence to Hela cells, survival in human macrophages and a Caenorhabditis elegans infection model. The aminoterminal portion of the EF3314 protein was overexpressed in E. coli to obtain mouse polyclonal antibodies for use in Western blotting and immunolocalization experiments. Results The EF3314 gene has an unusually high GC content (46.88% vs. an average of 37.5% in the E. faecalis chromosome) and encodes a protein of 1744 amino acids that presents a series of 14 imperfect repeats of 90 amino acids covering almost the entire length of the protein. Its global organization is similar to the alpha-like protein family of group B streptococci, enterococcal surface protein Esp and bio film associated protein Bap from S. aureus. The EF3314 gene was always present and specific for E. faecalis strains of human, food and animal origin. Differences in size depended on variable numbers of repeats in the repetitive region. Conclusions EF3314 is a newly described, surface exposed protein that contributes to the virulence properties of E. faecalis.

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