Cell surface-associated lipoteichoic acid acts as an adhesion factor for attachment of Lactobacillus johnsonii La1 to human enterocyte-like Caco-2 cells.
The influence of pH on the adhesion of two Lactobacillus strains to Caco-2 human intestinal cells was investigated. One strain, Lactobacillus johnsonii La1, was adherent at any pH between 4 and 7. The other one, L. acidophilus La10, did not attach to this cell line under the same experimental conditions. On the basis of these results, we used the monoclonal antibody technique as a tool to determine differences on the surface of these bacteria and to identify a factor for adhesion. Mice were immunized with live La1, and the hybridomas produced by fusion of spleen cells with ONS1 cells were screened for the production of antibodies specific for L. johnsonii La1. A set of these monoclonal antibodies was directed against a nonproteinaceous component of the L. johnsonii La1 surface. It was identified as lipoteichoic acid (LTA). This molecule was isolated, chemically characterized, and tested in adhesion experiments in the same system. The adhesion of L. johnsonii La1 to Caco-2 cells was inhibited in a concentration-dependent way by purified LTA as well as by L. johnsonii La1 culture supernatant that contained LTA. These results showed that the mechanism of adhesion of L. johnsonii La1 to human Caco-2 cells involves LTA.
Granato
D
D
Nestlé Research Center, CH-1000 Lausanne 26, Switzerland. dominiique.granato@chlsnr.nestrd.ch
Perotti
F
F
Masserey
I
I
Rouvet
M
M
Golliard
M
M
Servin
A
A
Brassart
D
D
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology; D001422:Bacterial Adhesion; D018938:Caco-2 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007114:Immunization; D007150:Immunohistochemistry; D007778:Lactobacillus / Q000737:chemistry / Q000502:physiology*; D007779:Lactobacillus acidophilus / Q000737:chemistry / Q000502:physiology; D008070:Lipopolysaccharides / Q000032:analysis / Q000378:metabolism*; D008565:Membrane Proteins / Q000276:immunology; D051379:Mice; D013682:Teichoic Acids / Q000032:analysis / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D013682:Teichoic Acids; C009900:lipoteichoic acid