title: Surface protein p104 is involved in adhesion of Listeria monocytogenes to human intestinal cell line, Caco-2.
abstract: Adhesion of Listeria monocytogenes to intestinal endothelial cells is an important initial event in the pathogenesis of infection which is not well understood. The suggestion has been made that some proteins, including internalin and actin polymerisation protein (ActA), and carbohydrate molecules mediate, at least in part, the adhesion of listeria to certain cultured mammalian cells. This study investigated the role of a L. monocytogenes cell-surface protein of 104 kDa (p104) in adhesion to human intestinal enterocyte-like Caco-2 cell lines by transposon (Tn916) mutagenesis and a p104-specific monoclonal antibody (MAb-H7). Genotypic and phenotypic characteristics of Tn916-transformed L. monocytogenes strains, AAMU530 and AAMU572, revealed that these strains did not express p104, and the transposon had been inserted at a single locus in the structural gene. Strains AAMU530 and AAMU572 yielded only 10 and 6.3% adhesion to Caco-2 cells. Coating of L. monocytogenes and L. innocua wild-type strains with MAb-H7 reduced adhesion to Caco-2 cells from 100% to 50 and 45%, respectively, whereas on isotype control MAb EM-7G1 had no effect. Western blot analysis with MAb-H7 indicated that p104 is present in all Listeria spp. except in L. grayi. Furthermore, p104 is also present in internalin (BUG8) and ActA (LUT12) deficient strains, suggesting that p104 is indeed different from internalin or ActA proteins. Cytotoxicity analysis of strains AAMU530 and AAMU572 demonstrated that these strains, although haemolytic and phospholipase-positive, were avirulent when tested with a hybridoma B-lymphocyte cell line. Loss of virulence could be attributed to the interruption of adhesion of mutant strains to the hybridoma cell line. These results strongly suggest that p104 is an adhesion factor in L. monocytogenes and possibly in other Listeria species and is involved in adhesion to intestinal cells.
authors.forename: V K
authors.initials: VK
authors.lastname: Pandiripally
authors.forename: D G
authors.initials: DG
authors.lastname: Westbrook
authors.forename: G R
authors.initials: GR
authors.lastname: Sunki
authors.forename: A K
authors.initials: AK
authors.lastname: Bhunia
mesh_terms: D001422:Bacterial Adhesion / Q000502:physiology*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000502:physiology*; D015139:Blotting, Southern; D015153:Blotting, Western; D018938:Caco-2 Cells / Q000382:microbiology*; D003227:Conjugation, Genetic; D003601:Cytotoxicity Tests, Immunologic; D003602:Cytotoxicity, Immunologic; D006801:Humans; D006825:Hybridomas; D008089:Listeria monocytogenes / Q000166:cytology* / Q000235:genetics; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction