Y Oh1, P Varmanen, X Y Han, G Bennett, Z Xu, T Lu, A Palva. 1. Department of Thoracic/Head & Neck Medical Oncology, UT M.D. Anderson Cancer Center, Houston, TX 77030, USA. ywoh@mdanderson.org
Abstract
AIMS: To evaluate strains of lactobacilli for their ability to persist and secrete heterologous protein in the oral cavity. METHODS AND RESULTS: Four different strains of common oral lactobacilli, Lactobacillus brevis, Lactobacillus johnsonii, Lactobacillus murinus and Lactobacillus plantarum, were transformed with the plasmid pKTH2121, which contains a secretion cassette for beta-lactamase. Lactobacilli isolated from the mouth of host mice were also transformed with pKTH2121 for later feeding. Lactococcus lactis, transformed with pKTH2121, was also fed to mice as a negative control. All transformed isolates were fed to C57Black mice in varying schedules. The number of transformed bacteria persisting in the mouth was reported as a percentage of total oral bacteria recovered by swabbing. CONCLUSIONS: The transformed L. lactis, L. brevis, L. johnsonii, L. murinus, and the endogenous murine lactobacillus strain failed to persist in the mouth. Transformed L. plantarum, however, persisted in the mouth and comprised up to 25% of the total lactobacilli at 18 h and 10% at 24 h after feeding. L. plantarum recovered after feeding retained its ability to secrete beta-lactamase into culture medium efficiently. Beta-lactamase activity could be detected in oral secretions at 8 h after feedings. After repeated feedings, however, the L. plantarum containing pKTH2121 gradually lost its ability to persist after feedings. This experiment demonstrates that L. plantarum can transiently colonize the oral mucosa in large numbers, while continuously secreting foreign proteins, raising the possibility of using lactobacilli as a vector for delivery of oral mucosal peptides.
AIMS: To evaluate strains of lactobacilli for their ability to persist and secrete heterologous protein in the oral cavity. METHODS AND RESULTS: Four different strains of common oral lactobacilli, Lactobacillus brevis, Lactobacillus johnsonii, Lactobacillus murinus and Lactobacillus plantarum, were transformed with the plasmid pKTH2121, which contains a secretion cassette for beta-lactamase. Lactobacilli isolated from the mouth of host mice were also transformed with pKTH2121 for later feeding. Lactococcus lactis, transformed with pKTH2121, was also fed to mice as a negative control. All transformed isolates were fed to C57Black mice in varying schedules. The number of transformed bacteria persisting in the mouth was reported as a percentage of total oral bacteria recovered by swabbing. CONCLUSIONS: The transformed L. lactis, L. brevis, L. johnsonii, L. murinus, and the endogenous murinelactobacillus strain failed to persist in the mouth. Transformed L. plantarum, however, persisted in the mouth and comprised up to 25% of the total lactobacilli at 18 h and 10% at 24 h after feeding. L. plantarum recovered after feeding retained its ability to secrete beta-lactamase into culture medium efficiently. Beta-lactamase activity could be detected in oral secretions at 8 h after feedings. After repeated feedings, however, the L. plantarum containing pKTH2121 gradually lost its ability to persist after feedings. This experiment demonstrates that L. plantarum can transiently colonize the oral mucosa in large numbers, while continuously secreting foreign proteins, raising the possibility of using lactobacilli as a vector for delivery of oral mucosal peptides.