BACKGROUND: Rotavirus-induced diarrhea poses a worldwide medical problem in causing substantial morbidity and mortality among children in developing countries. We therefore developed a system for passive immunotherapy in which recombinant lactobacilli constitutively express neutralizing variable domain of llama heavy-chain (VHH) antibody fragments against rotavirus. METHODS: VHH were expressed in Lactobacillus paracasei, in both secreted and cell surface-anchored forms. Electron microscopy was used to investigate the binding efficacy of VHH-expressing lactobacilli. To investigate the in vivo function of VHH-expressing lactobacilli, a mouse pup model of rotavirus infection was used. RESULTS: Efficient binding of the VHH antibody fragments to rotavirus was shown by enzyme-linked immunosorbent assay and scanning electron microscopy. VHH fragments expressed by lactobacilli conferred a significant reduction in infection in cell cultures. When administered orally, lactobacilli-producing surface-expressed VHH markedly shortened disease duration, severity, and viral load in a mouse model of rotavirus-induced diarrhea when administered both fresh and in a freeze-dried form. CONCLUSIONS: Transformed lactobacilli may form the basis of a novel form of prophylactic treatment against rotavirus infections and other diarrheal diseases.
BACKGROUND: Rotavirus-induced diarrhea poses a worldwide medical problem in causing substantial morbidity and mortality among children in developing countries. We therefore developed a system for passive immunotherapy in which recombinant lactobacilli constitutively express neutralizing variable domain of llama heavy-chain (VHH) antibody fragments against rotavirus. METHODS: VHH were expressed in Lactobacillus paracasei, in both secreted and cell surface-anchored forms. Electron microscopy was used to investigate the binding efficacy of VHH-expressing lactobacilli. To investigate the in vivo function of VHH-expressing lactobacilli, a mouse pup model of rotavirus infection was used. RESULTS: Efficient binding of the VHH antibody fragments to rotavirus was shown by enzyme-linked immunosorbent assay and scanning electron microscopy. VHH fragments expressed by lactobacilli conferred a significant reduction in infection in cell cultures. When administered orally, lactobacilli-producing surface-expressed VHH markedly shortened disease duration, severity, and viral load in a mouse model of rotavirus-induced diarrhea when administered both fresh and in a freeze-dried form. CONCLUSIONS: Transformed lactobacilli may form the basis of a novel form of prophylactic treatment against rotavirus infections and other diarrheal diseases.
Authors: Luciano J Melli; Vanesa Zylberman; Yanina Hiriart; Constanza E Lauche; Ariela Baschkier; Romina Pardo; Elizabeth Miliwebsky; Isabel Chinen; Marta Rivas; Fernando A Goldbaum; Juan E Ugalde; Diego J Comerci; Andrés E Ciocchini Journal: J Clin Microbiol Date: 2020-02-24 Impact factor: 5.948
Authors: M Cruz Martín; Neha Pant; Victor Ladero; Gökçe Günaydin; Kasper Krogh Andersen; Beatriz Alvarez; Noelia Martínez; Miguel A Alvarez; Lennart Hammarström; Harold Marcotte Journal: Appl Environ Microbiol Date: 2011-01-21 Impact factor: 4.792
Authors: Janusz Wesolowski; Vanina Alzogaray; Jan Reyelt; Mandy Unger; Karla Juarez; Mariela Urrutia; Ana Cauerhff; Welbeck Danquah; Björn Rissiek; Felix Scheuplein; Nicole Schwarz; Sahil Adriouch; Olivier Boyer; Michel Seman; Alexei Licea; David V Serreze; Fernando A Goldbaum; Friedrich Haag; Friedrich Koch-Nolte Journal: Med Microbiol Immunol Date: 2009-06-16 Impact factor: 3.402