Site-specific Alterations in the B Oligomer that Affect Receptor-binding Activities and Mitogenicity of Pertussis Toxin

Lobet, Yves; Feron, Christiane; Dequesne, Guy; Simoen, E; Hauser, P; Locht, Camille

doi:10.1006/biol.1993.1042

Cited by 10 publications

(16 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The ptx genes harboring mutations leading to the R9K and E129G substitutions in the S1 subunit of PT were introduced by homologous recombination into the chromosome of BPDR, resulting in the BPDR-RE strain. Both substitutions abolish the ADP-ribosyltransferase activity of PT (15). Immunoblot analysis of culture supernatants showed that PT-RE is efficiently produced and secreted by BPDR-RE (data not shown).…”

Section: Resultsmentioning

confidence: 95%

“…B. pertussis BPDR-RE was constructed by introducing pPT-RE into the chromosome of BPDR by a single homologous recombination event. pPT-RE is a pPT2 derivative (1), containing the PT genes with mutations resulting in the R9K and E129G substitutions in the S1 subunit (15). The recombinant strain was selected on BG plates containing streptomycin and ampicillin.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Role of ADP-Ribosyltransferase Activity of Pertussis Toxin in Toxin-Adhesin Redundancy with Filamentous Hemagglutinin duringBordetella pertussisInfection

et al. 2001

Self Cite

View full text Add to dashboard Cite

Pertussis toxin (PT) and filamentous hemagglutinin (FHA) are two major virulence factors of Bordetella pertussis. FHA is the main adhesin, whereas PT is a toxin with an A-B structure, in which the A protomer expresses ADP-ribosyltransferase activity and the B moiety is responsible for binding to the target cells. Here, we show redundancy of FHA and PT during infection. Whereas PT-deficient and FHA-deficient mutants colonized the mouse respiratory tract nearly as efficiently as did the isogenic parent strain, a mutant deficient for both factors colonized substantially less well. This was not due to redundant functions of PT and FHA as adhesins, since in vitro studies of epithelial cells and macrophages indicated that FHA, but not PT, acts as an adhesin. An FHA-deficient B. pertussis strain producing enzymatically inactive PT colonized as poorly as did the FHAdeficient, PT-deficient strain, indicating that the ADP-ribosyltransferase activity of PT is required for redundancy with FHA. Only strains producing active PT induced a local transient release of tumor necrosis factor alpha (TNF-␣), suggesting that the pharmacological effects of PT are the basis of the redundancy with FHA, through the release of TNF-␣. This may lead to damage of the pulmonary epithelium, allowing the bacteria to colonize even in the absence of FHA.Many bacterial pathogens produce adhesins and toxins to express their pathogenic potential. Although they are often studied as separate entities, it is possible that in some instances they act in a synergistic or in a redundant way. Redundancy among adhesins has previously been shown (40); however, redundancy between a toxin and an adhesin has not yet been demonstrated. Bordetella pertussis is a human pathogen that causes whooping cough after adherence to and colonization of the respiratory tract. B. pertussis synthesizes a number of virulence factors (16), whose production is under the control of a regulatory locus named bvg. Among them, filamentous hemagglutinin (FHA) and pertussis toxin (PT) have been proven to protect against bacterial infection (4,31,35) and are part of the present acellular vaccines. FHA is a monomeric protein of 220 kDa and has a filamentous structure (20). It is both surface exposed and secreted by B. pertussis. FHA is considered to be the main adhesin of B. pertussis (18) and contains at least three distinct binding sites: a glycosaminoglycan and a carbohydrate binding sites (10, 27) and an arginine-glycine-aspartate (RGD) sequence involved in binding to the CR3 integrin of macrophages (28). PT is an ADP-ribosylating toxin, belonging to the AB 5 family of toxins (17) and is secreted into the extracellular milieu. PT consists of five subunits, named S1 to S5, and can be divided into two functional moieties. The A protomer, corresponding to the S1 subunit, expresses the ADP-ribosyltransferase activity. The B oligomer, composed of subunits S2 to S5, mediates binding of PT to its target cell receptors through the carbohydrate recognition domains of the S2 and S3 subunits (43)....

show abstract

Section: Resultsmentioning

confidence: 95%

Section: Methodsmentioning

confidence: 99%

Role of ADP-Ribosyltransferase Activity of Pertussis Toxin in Toxin-Adhesin Redundancy with Filamentous Hemagglutinin duringBordetella pertussisInfection

et al. 2001

Self Cite

View full text Add to dashboard Cite

show abstract

“…Purified pertussis toxin (PTX-R), provided by the Pasteur Institute (Lille, France) (20), and filamentous hemagglutinin (FHA), 3 provided by Chiron Biocine (Sienna, Italy), were used for in vitro stimulation. The enzymatically inactive form PTX-R was used instead of natural pertussis toxin to avoid potential interference by the toxin-expressed ADP ribosylation of signal-transducing proteins in the target cells.…”

Section: B Pertussis Ags Used For In Vitro Stimulationmentioning

confidence: 99%

Bordetella pertussisInfection in 2-Month-Old Infants Promotes Type 1 T Cell Responses

Mascart

Verscheure

Malfroot

et al. 2003

The Journal of Immunology

Self Cite

126

View full text Add to dashboard Cite

Neonatal immaturity of the immune system is currently believed to generally limit the induction of immune responses to vaccine Ags and to skew them toward type 2 responses. We demonstrated here that Bordetella pertussis infection in very young infants (median, 2 mo old) as well as the first administration of whole-cell pertussis vaccine induces B. pertussis Ag-specific IFN-γ secretion by the PBMC of these infants. IFN-γ was secreted by both CD4+ and CD8+ T lymphocytes, and the levels of Ag-induced IFN-γ secretion did not correlate with the age of the infants. Appearance of the specific Th-1 cell-mediated immunity was accompanied by a general shift of the cytokine secretion profile of these infants toward a stronger Th1 profile, as evidenced by the response to a polyclonal stimulation. We conclude that the immune system of 2-mo-old infants is developmentally mature enough to develop Th1 responses in vivo upon infection by B. pertussis or vaccination with whole-cell pertussis vaccines.

show abstract

“…2b), suggesting that the THP-PTX interaction is mediated by this oligomer. PTX (Lobet et al, 1993), we tested the THP-PTX interactions using PTX produced by B. pertussis mutants which were unable to produce the S2 or S3 subunit (∆S2 or ∆S3 mutant, respectively). [These mutants do, however, produce a correctly assembled holotoxin, through replacement of the missing subunit by a second copy of the S3 or S2 subunit (D. Raze, F. D. Menozzi, R. Antoine, Y. Sato and C. Locht, unpublished)].…”

Section: Ptx Binds To Thp Via the S2 Subunit Of The B Oligomermentioning

confidence: 99%

“…This indicates that the S2 subunit is involved in the recognition of THP. The residual THP binding observed with the PTX ∆S2 mutant is likely due to the high amino acid similarity displayed by the S2 and S3 subunits (Lobet et al, 1993).…”

Section: Ptx Binds To Thp Via the S2 Subunit Of The B Oligomermentioning

confidence: 99%

Interaction of human Tamm–Horsfall glycoprotein with Bordetella pertussis toxin

Menozzi¹,

Debrie²,

Tissier³

et al. 2002

Microbiology

Self Cite

View full text Add to dashboard Cite

Tamm-Horsfall glycoprotein (THP), which is synthesized by renal tubular cells, is the most abundant protein in normal human urine. Although its physiological function remains unclear, it has been proposed that THP may act as a defence factor against urinary tract infections by inhibiting the binding of S-and P-fimbriated Escherichia coli to renal epithelial cells. Because THPrelated proteins are also found in the superficial layers of the oral mucosa, the authors investigated the ability of THP to interfere with the cytoadherence of pathogenic bacteria that colonize mucosal surfaces other than those of the urogenital tract. In this report, it is shown that THP binds to virulent Bordetella pertussis and reduces its adherence to both renal and pulmonary epithelial cells. This cytoadherence inhibitory effect was not observed with a B. pertussis mutant lacking the pertussis toxin (PTX) operon, and was dependent on the direct interaction of THP with the S2 subunit within the PTX B oligomer. The authors also show that the glycosylation moiety of THP is crucial for its binding to PTX. The THP-PTX interaction was exploited to develop an affinity chromatography method that allows a one-step purification of active PTX. These observations suggest that besides its anti-adherence activity, THP may also trap toxins produced by pathogenic bacteria that colonize mucosal surfaces.

show abstract

Site-specific Alterations in the B Oligomer that Affect Receptor-binding Activities and Mitogenicity of Pertussis Toxin

Cited by 10 publications

References 0 publications

Role of ADP-Ribosyltransferase Activity of Pertussis Toxin in Toxin-Adhesin Redundancy with Filamentous Hemagglutinin duringBordetella pertussisInfection

Role of ADP-Ribosyltransferase Activity of Pertussis Toxin in Toxin-Adhesin Redundancy with Filamentous Hemagglutinin duringBordetella pertussisInfection

Bordetella pertussisInfection in 2-Month-Old Infants Promotes Type 1 T Cell Responses

Interaction of human Tamm–Horsfall glycoprotein with Bordetella pertussis toxin

Contact Info

Product

Resources

About