Helicobacter pylori-antigen-binding fragments expressed on the filamentous M13 phage prevent bacterial growth

• Published in: Biochimica et biophysica acta Vol. 1474; no. 1; pp. 107 - 113
• Main Authors: Cao, Jun, Sun, Yi-qian, Berglindh, Thomas, Mellgård, Björn, Li, Zhao-qi, Mårdh, Bibbi, Mårdh, Sven
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 06.03.2000
• Subjects: Animals; Antibodies, Monoclonal - immunology; Bactericidal; Bacteriophage M13 - genetics; Bacteriophage M13 - immunology; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Genetic Engineering; Helicobacter Infections - prevention & control; Helicobacter pylori - genetics; Helicobacter pylori - growth & development; Helicobacter pylori - immunology; Helicobacter pylori infection; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - immunology; MEDICIN; MEDICINE; Mice; Microscopy, Fluorescence; Monoclonal antibody; Mouse model; Phage specificity; Recombinant bacteriophage; Single-chain fragment variable (ScFv); Single-chain fragment variable (ScFv); Phage specificity; Helicobacter pylori infection; Bactericidal; Mouse model; Monoclonal antibody; Recombinant bacteriophage
• ISSN: 0304-4165; 0006-3002; 1872-8006; 1872-8006
• DOI: 10.1016/S0304-4165(00)00005-2

Colonization of the human stomach by Helicobacter pylori is associated with the development of gastritis, duodenal ulcer, mucosa-associated lymphoid tissue (MALT) lymphoma, and gastric cancer. H. pylori-antigen-binding single-chain variable fragments (ScFv) were derived from murine hybridomas producing monoclonal antibodies and expressed as a g3p-fusion protein on a filamentous M13 phage. The recombinant ScFv-phage reacted specifically with a 30-kDa monomeric protein of a H. pylori surface antigen preparation and by means of immunofluorescence microscopy the phage was shown to bind to both the spiral and coccoid forms of the bacterium. In vitro, the recombinant phage exhibited a bacteriocidal effect and inhibited specifically the growth of all the six strains of H. pylori tested. When H. pylori was pretreated with the phage 10 min before oral inoculation of mice, the colonization of the mouse stomachs by the bacterium was significantly reduced ( P<0.01). The results suggest that genetic engineering may be used to generate therapy-effective phages.