Prospective uses of recombinant Lactococcus lactis expressing both listeriolysin O and mutated internalin A from Listeria monocytogenes as a tool for DNA vaccination

• Published in: Genetics and molecular research Vol. 14; no. 4; pp. 18485 - 18493
• Main Authors: De Azevedo, M S P, Santos Rocha, C, Pereira, V B, De Oliveira Junior, A F, De Sousa, C S, Azevedo, V, LeBlanc, J G, Chatel, J M, Miyoshi, A
• Format: Journal Article
• Language: English
• Published: Brazil Ribeirão Preto foundation for Scientific Research (FUNPEC) 01-01-2015
• Subjects: Bacterial Proteins - genetics; Bacterial Proteins - immunology; Bacterial Toxins - genetics; Bacterial Toxins - immunology; Bacterial Vaccines; Cell Membrane - metabolism; Gene Expression; Heat-Shock Proteins - genetics; Heat-Shock Proteins - immunology; Hemolysin Proteins - genetics; Hemolysin Proteins - immunology; Hemolysis; Lactococcus lactis; Lactococcus lactis - genetics; Lactococcus lactis - metabolism; Life Sciences; Listeria monocytogenes; Listeria monocytogenes - immunology; Mutation; Recombinant Proteins; Vaccination; listériolysine; listeria monocytogènes; Listeriolysin O; Mutated internalin A; lactococcus lactis; internaline; lactic acid bacteria; bactérie lactique
• ISSN: 1676-5680; 1676-5680
• DOI: 10.4238/2015.December.23.36

In this study, Lactococcus lactis was engineered to express mutated internalin A on its surface and to secrete large amounts of listeriolysin O (LLO) in order to improve its potential as a vehicle for DNA vaccination. Western blotting experiments demonstrated that the bacterium expressed LLO in both the cytoplasmic and extracellular compartments, with higher quantities found in the culture supernatants. A hemolytic assay showed that the recombinant strain secreted 250 ng active LLO/mg total protein. This mInlA/LLO-producing strain of L. lactis may be used as an alternative tool in DNA vaccination against a number of infectious diseases or in cancer therapy.