Interaction of the Bordetella pertussis filamentous hemagglutinin with heparin

Abstract

Heparin, a glycosaminoglycan synthesized in connective tissue-mast cells, appeared to inhibit the hemagglutination of rabbit erythrocytes induced by the filamentous hemagglutinin (FHA), a major adhesin of Bordetella pertussis. This inhibition suggested an interaction of heparin with the FHA region responsible for the hemagglutination activity. FHA-heparin interactions may play a role in bacterial attachment and persistence in the lungs during human pertussis. To confirm a direct FHA-heparin interaction, heparin was used as ligand in an affinity chromatography procedure. This technique allowed to purify FHA directly from the bacterial culture medium in a single-step using heparin-Sepharose CL-6B or Zetaffinity heparin 60 disks. The purified FHA was highly immunoreactive with anti-FHA monoclonal antibodies and showed no signs of degradation after 15 successive cycles of freezing-thawing. The described purification method is simple, and suitable for the rapid preparation of FHA.

Molecular characterization of Bordetella bronchiseptica filamentous haemagglutinin and its secretion machinery The GenBank accession numbers for the sequences reported in this paper are AF111794, AF111796, AF111797 and AF111798.

“…B. bronchiseptica GP1 was originally isolated from a guinea pig (Akerley et al, 1992). B. bronchiseptica BB1015 (Menozzi et al, 1991), RB50 (Cotter & Miller, 1994), NL1013, S87, 899L and NL1011 (Antoine & Locht, 1992) and B. pertussis BP536 (Relman et al, 1989), BPSM (Menozzi et al, 1994a), BPGR4 (∆fhaB) and BPEC (∆fhaC) (Gue! din et al, 1998) (Nordmann et al, 1992) and B. parapertussis 8234 was a gift from E. Hewlett, University of Virginia.…”

“…Polyclonal chicken anti-FHA IgY and polyclonal anti-FhaC rat IgG were made by Eurogentec. FHA and Fha44 were purified from BPGR4 culture supernatants by heparin-Sepharose chromatography as described by Menozzi et al (1991). Membrane extracts for the detection of FhaC were prepared as described by Jacob-Dubuisson et al (1996) except that the Sarkosyl extraction step was omitted.…”

“…FHA is recognized by lactose-containing glycolipids on ciliated respiratory epithelial cells (Tuomanen et al, 1988). It binds to sulphated carbohydrates of sulphatides and proteoglycans on the surface of epithelial cells or in the extracellular matrix (Brennan et al, 1991 ;Menozzi et al, 1991Menozzi et al, , 1994a. In addition, it possesses an RGD motif, which is a canonical recognition sequence for members of the integrin family.…”

Enhanced bacterial virulence through exploitation of host glycosaminoglycans

“…This suggested that B. pertussis may attach to GAGs, which are found in abundance in the mucosa of the bronchial airways. Menozzi et al (1991) demonstrated that FHA binds to heparin, a property that can be utilized to purify the protein, and that the heparin-inhibitable attachment of B. pertussis to epithelial cells is mediated largely by FHA . FHA binds to the same sulphated glycolipids as virulent B. pertussis, and heparan sulphate proteoglycans have been identified as FHA receptors on epithelial cells .…”

Role of Bordetella bronchiseptica adenylate cyclase in nasal colonization and in development of local and systemic immune responses in piglets

“…infection with B. bronchiseptica, mice develop anti-FHA antibodies in the serum and in the bronchoalveolar lavage fluid [12,13]. B. bronchiseptica FHA has a molecular mass and hemagglutination properties that are similar to those of B. pertussis FHA and the two strains share the same epitopes [18,27,28]. As in mice, anti-B.…”