title: Adherence of Brucella to human epithelial cells and macrophages is mediated by sialic acid residues.
abstract: The basis for the interaction of Brucella species with the surface of epithelial cells before migration in the host within polymorphonuclear leucocytes is largely unknown. Here, we studied the ability of Brucella abortus and Brucella melitensis to adhere to cultured epithelial (HeLa and HEp-2) cells and THP-1-derived macrophages, and to bind extracellular matrix proteins (ECM). The brucellae adhered to epithelial cells forming localized bacterial microcolonies on the cell surface, and this process was inhibited significantly by pretreatment of epithelial cells with neuraminidase and sodium periodate and by preincubation of the bacteria with heparan sulphate and N-acetylneuraminic acid. Trypsinization of epithelial cells yielded increased adherence, suggesting unmasking of target sites on host cells. Notably, the brucellae also adhered to cultured THP-1 cells, and this event was greatly reduced upon removal of sialic acid residues from these cells with neuraminidase. B. abortus bound in a dose-dependent manner to immobilized fibronectin and vitronectin and, to a lesser extent, to chondroitin sulphate, collagen and laminin. In sum, our data strongly suggest that the adherence mechanism of brucellae to epithelial cells and macrophages is mediated by cellular receptors containing sialic acid and sulphated residues. The recognition of ECM (fibronectin and vitronectin) by the brucellae may represent a mechanism for spread within the host tissues. These are novel findings that offer new insights into understanding the interplay between Brucella and host cells.
authors.affiliation: Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Edificio 76, Complejo de Ciencias, Puebla, México.
authors.forename: Elsa I
authors.initials: EI
authors.lastname: Castañeda-Roldán
authors.forename: Fabiola
authors.initials: F
authors.lastname: Avelino-Flores
authors.forename: Monique
authors.initials: M
authors.lastname: Dall'Agnol
authors.forename: Enrique
authors.initials: E
authors.lastname: Freer
authors.forename: Lilia
authors.initials: L
authors.lastname: Cedillo
authors.forename: Jacques
authors.initials: J
authors.lastname: Dornand
authors.forename: Jorge A
authors.initials: JA
authors.lastname: Girón
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion / Q000502:physiology*; D002003:Brucella abortus / Q000502:physiology* / Q000648:ultrastructure; D017347:Brucella melitensis / Q000378:metabolism* / Q000648:ultrastructure; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology* / Q000648:ultrastructure; D016326:Extracellular Matrix Proteins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000378:metabolism* / Q000382:microbiology; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D009439:Neuraminidase / Q000378:metabolism; D010504:Periodic Acid / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D016326:Extracellular Matrix Proteins; D010504:Periodic Acid; C009288:metaperiodate; D009439:Neuraminidase; D014357:Trypsin; D019158:N-Acetylneuraminic Acid