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Oral Delivery of a Novel Recombinant Streptococcus mitis Vector Elicits Robust Vaccine Antigen-Specific Oral Mucosal and Systemic Antibody Responses and T Cell Tolerance.

Xie E, Kotha A, Biaco T, Sedani N, Zou J, Stashenko P, Duncan MJ, Campos-Neto A, Cayabyab MJ - PLoS ONE (2015)

Bottom Line: Oral vaccination led to the efficient and persistent bacterial colonization of the mouth and the induction of both salivary and systemic antibody responses.Interestingly, persistently colonized animals developed antigen-specific systemic T cell tolerance.Based on these findings we propose the use of rS. mitis vaccine vector for the induction of mucosal antibodies that will prevent the penetration of the mucosa by pathogens such as HIV.

View Article: PubMed Central - PubMed

Affiliation: Global Infectious Disease Research Center and the Department of Immunology and Infectious Diseases, The Forsyth Institute, 245 First Street, Cambridge, Massachusetts, United States of America.

ABSTRACT
The pioneer human oral commensal bacterium Streptococcus mitis has unique biologic features that make it an attractive mucosal vaccine or therapeutic delivery vector. S. mitis is safe as a natural persistent colonizer of the mouth, throat and nasopharynx and the oral commensal bacterium is capable of inducing mucosal antibody responses. A recombinant S. mitis (rS. mitis) that stably expresses HIV envelope protein was generated and tested in the germ-free mouse model to evaluate the potential usefulness of this vector as a mucosal vaccine against HIV. Oral vaccination led to the efficient and persistent bacterial colonization of the mouth and the induction of both salivary and systemic antibody responses. Interestingly, persistently colonized animals developed antigen-specific systemic T cell tolerance. Based on these findings we propose the use of rS. mitis vaccine vector for the induction of mucosal antibodies that will prevent the penetration of the mucosa by pathogens such as HIV. Moreover, the first demonstration of rS. mitis having the ability to elicit T cell tolerance suggest the potential use of rS. mitis as an immunotherapeutic vector to treat inflammatory, allergic and autoimmune diseases.

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Construction of recombinant S. mitis expressing HIV Env gp120.(A) Recombinant S. mitis (rS. mitis) was generated by homologous recombination. The p5E3 suicide plasmid was created by inserting the S. mitis codon-optimized HIV-1 HXBc2 Env gp120-His-tag (HIV Env) in-frame with the 250bp 5’ end of the pullulanase gene (pulA/Smt0163) followed by ermr gene and the 250bp 3’ end of the pulA gene into pCR2.1 for transformation and integration into the Smt0163 locus. (B) S. mitis was transformed with water (control), p5E3 (Smt0163:HIVenv:ermr) or pCR2.1 containing ermr flanked by 250bp pulA 5’ and 3’ fragments (Smt0163:ermr). Growth of transformants on THB plates containing 50μg/ml erythromycin and transformation frequency are shown. (C) Integration was confirmed by PCR using S. mitis-specific primers A/B, HIV-specific primer C and ermr-specific primer D.
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pone.0143422.g001: Construction of recombinant S. mitis expressing HIV Env gp120.(A) Recombinant S. mitis (rS. mitis) was generated by homologous recombination. The p5E3 suicide plasmid was created by inserting the S. mitis codon-optimized HIV-1 HXBc2 Env gp120-His-tag (HIV Env) in-frame with the 250bp 5’ end of the pullulanase gene (pulA/Smt0163) followed by ermr gene and the 250bp 3’ end of the pulA gene into pCR2.1 for transformation and integration into the Smt0163 locus. (B) S. mitis was transformed with water (control), p5E3 (Smt0163:HIVenv:ermr) or pCR2.1 containing ermr flanked by 250bp pulA 5’ and 3’ fragments (Smt0163:ermr). Growth of transformants on THB plates containing 50μg/ml erythromycin and transformation frequency are shown. (C) Integration was confirmed by PCR using S. mitis-specific primers A/B, HIV-specific primer C and ermr-specific primer D.

Mentions: Recombinant S. mitis (rS. mitis) was generated by homologous recombination. The p5E3 suicide plasmid vector was created by inserting HIV-1 HXBc2 Env gp120-His-tag (HIV Env) [14] (codon-optimized for S. mitis expression by BlueHeron Technology, Bothel, WA) in-frame with the 250bp 5’ end of the pullulanase gene (pulA/Smt0163) followed by Ermr gene [15] and the 250bp 3’ end of the pulA gene into pCR2.1 (Invitrogen, Carlsbad, CA) for transformation and integration into the Smt0163 locus (Fig 1A). S. mitis was transformed by electroporation with p5E3 to generate homologous recombinant S. mitis expressing HIV-1 Env or with pCR2.1 containing Ermr flanked by 250bp pulA 5’ and 3’ fragments to generate control S. mitis empty vector. rS. mitis clones were selected on Todd Hewitt Broth (THB) plates containing 50 μg/ml erythromycin (Fisher Scientific, Pittsburgh, PA).


Oral Delivery of a Novel Recombinant Streptococcus mitis Vector Elicits Robust Vaccine Antigen-Specific Oral Mucosal and Systemic Antibody Responses and T Cell Tolerance.

Xie E, Kotha A, Biaco T, Sedani N, Zou J, Stashenko P, Duncan MJ, Campos-Neto A, Cayabyab MJ - PLoS ONE (2015)

Construction of recombinant S. mitis expressing HIV Env gp120.(A) Recombinant S. mitis (rS. mitis) was generated by homologous recombination. The p5E3 suicide plasmid was created by inserting the S. mitis codon-optimized HIV-1 HXBc2 Env gp120-His-tag (HIV Env) in-frame with the 250bp 5’ end of the pullulanase gene (pulA/Smt0163) followed by ermr gene and the 250bp 3’ end of the pulA gene into pCR2.1 for transformation and integration into the Smt0163 locus. (B) S. mitis was transformed with water (control), p5E3 (Smt0163:HIVenv:ermr) or pCR2.1 containing ermr flanked by 250bp pulA 5’ and 3’ fragments (Smt0163:ermr). Growth of transformants on THB plates containing 50μg/ml erythromycin and transformation frequency are shown. (C) Integration was confirmed by PCR using S. mitis-specific primers A/B, HIV-specific primer C and ermr-specific primer D.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4664415&req=5

pone.0143422.g001: Construction of recombinant S. mitis expressing HIV Env gp120.(A) Recombinant S. mitis (rS. mitis) was generated by homologous recombination. The p5E3 suicide plasmid was created by inserting the S. mitis codon-optimized HIV-1 HXBc2 Env gp120-His-tag (HIV Env) in-frame with the 250bp 5’ end of the pullulanase gene (pulA/Smt0163) followed by ermr gene and the 250bp 3’ end of the pulA gene into pCR2.1 for transformation and integration into the Smt0163 locus. (B) S. mitis was transformed with water (control), p5E3 (Smt0163:HIVenv:ermr) or pCR2.1 containing ermr flanked by 250bp pulA 5’ and 3’ fragments (Smt0163:ermr). Growth of transformants on THB plates containing 50μg/ml erythromycin and transformation frequency are shown. (C) Integration was confirmed by PCR using S. mitis-specific primers A/B, HIV-specific primer C and ermr-specific primer D.
Mentions: Recombinant S. mitis (rS. mitis) was generated by homologous recombination. The p5E3 suicide plasmid vector was created by inserting HIV-1 HXBc2 Env gp120-His-tag (HIV Env) [14] (codon-optimized for S. mitis expression by BlueHeron Technology, Bothel, WA) in-frame with the 250bp 5’ end of the pullulanase gene (pulA/Smt0163) followed by Ermr gene [15] and the 250bp 3’ end of the pulA gene into pCR2.1 (Invitrogen, Carlsbad, CA) for transformation and integration into the Smt0163 locus (Fig 1A). S. mitis was transformed by electroporation with p5E3 to generate homologous recombinant S. mitis expressing HIV-1 Env or with pCR2.1 containing Ermr flanked by 250bp pulA 5’ and 3’ fragments to generate control S. mitis empty vector. rS. mitis clones were selected on Todd Hewitt Broth (THB) plates containing 50 μg/ml erythromycin (Fisher Scientific, Pittsburgh, PA).

Bottom Line: Oral vaccination led to the efficient and persistent bacterial colonization of the mouth and the induction of both salivary and systemic antibody responses.Interestingly, persistently colonized animals developed antigen-specific systemic T cell tolerance.Based on these findings we propose the use of rS. mitis vaccine vector for the induction of mucosal antibodies that will prevent the penetration of the mucosa by pathogens such as HIV.

View Article: PubMed Central - PubMed

Affiliation: Global Infectious Disease Research Center and the Department of Immunology and Infectious Diseases, The Forsyth Institute, 245 First Street, Cambridge, Massachusetts, United States of America.

ABSTRACT
The pioneer human oral commensal bacterium Streptococcus mitis has unique biologic features that make it an attractive mucosal vaccine or therapeutic delivery vector. S. mitis is safe as a natural persistent colonizer of the mouth, throat and nasopharynx and the oral commensal bacterium is capable of inducing mucosal antibody responses. A recombinant S. mitis (rS. mitis) that stably expresses HIV envelope protein was generated and tested in the germ-free mouse model to evaluate the potential usefulness of this vector as a mucosal vaccine against HIV. Oral vaccination led to the efficient and persistent bacterial colonization of the mouth and the induction of both salivary and systemic antibody responses. Interestingly, persistently colonized animals developed antigen-specific systemic T cell tolerance. Based on these findings we propose the use of rS. mitis vaccine vector for the induction of mucosal antibodies that will prevent the penetration of the mucosa by pathogens such as HIV. Moreover, the first demonstration of rS. mitis having the ability to elicit T cell tolerance suggest the potential use of rS. mitis as an immunotherapeutic vector to treat inflammatory, allergic and autoimmune diseases.

Show MeSH
Related in: MedlinePlus