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Lactobacilli expressing variable domain of llama heavy-chain antibody fragments (lactobodies) confer protection against rotavirus-induced diarrhea

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Author: Pant, N. · Hultberg, A. · Zhao, Y. · Svensson, L. · Pan-Hammarström, Q. · Johansen, K. · Pouwels, P.H. · Ruggeri, F.M. · Hermans, P. · Frenken, L. · Borén, T. · Marcotte, H. · Hammarström, L.
Type:article
Date:2006
Institution: TNO Preventie en Gezondheid
Source:Journal of Infectious Diseases, 11, 194, 1580-1588
Identifier: 239669
doi: doi:10.1086/508747
Keywords: Biology · Antibody · Immunoglobulin heavy chain · Animal experiment · Animal model · Animal tissue · Artiodactyla · Binding affinity · Cell culture · Controlled study · Disease duration · Disease severity · Electron microscopy · Enzyme linked immunosorbent assay · Immunity · Infectious diarrhea · Lactobacillus paracasei · Lama · Mouse · Nonhuman · Nucleotide sequence · Scanning electron microscope · Virus load · Virus neutralization · Animals · Antibodies, Viral · Camelids, New World · Diarrhea · Disease Models, Animal · Enzyme-Linked Immunosorbent Assay · Feces · Genetic Vectors · Immunization, Passive · Immunoglobulin Heavy Chains · Immunoglobulin Variable Region · Lactobacillus · Mice · Mice, Inbred BALB C · Microscopy, Electron, Scanning · Protein Binding · Rotavirus · Rotavirus Infections

Abstract

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. © 2006 by the Infectious Diseases Society of America. All rights reserved. Chemicals / CAS: Antibodies, Viral; Immunoglobulin Heavy Chains; Immunoglobulin Variable Region