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Infection and Immunity, June 2008, p. 2696-2705, Vol. 76, No. 6
0019-9567/08/$08.00+0     doi:10.1128/IAI.00119-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Nasal Immunization with Lactococcus lactis Expressing the Pneumococcal Protective Protein A Induces Protective Immunity in Mice{triangledown}

Marcela Medina,1 Julio Villena,1,2 Elisa Vintiñi,1 Elvira María Hebert,3 Raúl Raya,3* and Susana Alvarez1,2*

Laboratorio de Bioquímica y Clínica Experimental, Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, Tucumán, Argentina,1 Instituto de Bioquímica Aplicada, Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Balcarce 747, CP 4000, San Miguel de Tucumán, Tucumán, Argentina,2 Laboratorio de Genética y Biología Molecular, Centro de Referencia para Lactobacilos (CERELA-CONICET), Chacabuco 145, Tucumán, Argentina3

Received 28 January 2008/ Returned for modification 4 March 2008/ Accepted 31 March 2008

Nisin-controlled gene expression was used to develop a recombinant strain of Lactococcus lactis that is able to express the pneumococcal protective protein A (PppA) on its surface. Immunodetection assays confirmed that after the induction with nisin, the PppA antigen was predictably and efficiently displayed on the cell surface of the recombinant strain, which was termed L. lactis PppA. The production of mucosal and systemically specific antibodies in adult and young mice was evaluated after mice were nasally immunized with L. lactis PppA. Immunoglobulin M (IgM), IgG, and IgA anti-PppA antibodies were detected in the serum and bronchoalveolar lavage fluid of adult and young mice, which showed that PppA expressed in L. lactis was able to induce a strong mucosal and systemic immune response. Challenge survival experiments demonstrated that immunization with L. lactis PppA was able to increase resistance to systemic and respiratory infection with different pneumococcal serotypes, and passive immunization assays of naïve young mice demonstrated a direct correlation between anti-PppA antibodies and protection. The results presented in this study demonstrate three major characteristics of the effectiveness of nasal immunization with PppA expressed as a protein anchored to the cell wall of L. lactis: it elicited cross-protective immunity against different pneumococcal serotypes, it afforded protection against both systemic and respiratory challenges, and it induced protective immunity in mice of different ages.

* Corresponding author. Mailing address for Susana Alvarez: Laboratorio de Bioquímica y Clínica Experimental, CERELA-CONICET, Chacabuco 145, Tucumán, Argentina. Phone: 51-381-4310465. Fax: 51-381-4005600. E-mail: salvarez{at}cerela.org.ar. Mailing address for Raúl Raya: Laboratorio de Genética y Biología Molecular, CERELA- CONICET, Chacabuco 145, Tucumán, Argentina. Phone: 51-381- 4310465. Fax: 51-381-4005600. E-mail: rraya{at}cerela.org.ar

{triangledown} Published ahead of print on 7 April 2008.

Editor: R. P. Morrison

Infection and Immunity, June 2008, p. 2696-2705, Vol. 76, No. 6
0019-9567/08/$08.00+0     doi:10.1128/IAI.00119-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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