Responses of <i>Haemophilus pleuropneumoniae</i> to iron restriction: changes in the outer membrane protein profile and the removal of iron from porcine transferrin

Niven, Donald F.; Donga, J.; Archibald, Frederick

doi:10.1111/j.1365-2958.1989.tb00258.x

Cited by 52 publications

(41 citation statements)

References 42 publications

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“…The serum directed against the ExbB protein was raised in rabbits by an initial intracutaneous injection and two subcutaneous boost injections of 100 g of dissolved recombinant glutathione S-transferase (GST)-ExbB fusion protein in Emulsigen-Plus (MVP Inc., Ralston, Nebr.). Porcine transferrin was prepared by ammonium sulfate precipitation and subsequent column chromatography using DEAE-Sepharose CL-6B (Pharmacia, Freiburg, Germany) (24).…”

Section: Methodsmentioning

confidence: 99%

Actinobacillus pleuropneumoniaeIron Transport: a Set ofexbBDGenes Is Transcriptionally Linked to thetbpBGene and Required for Utilization of Transferrin-Bound Iron

et al. 2000

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Upon iron restriction, Actinobacillus pleuropneumoniae has been shown to express the transferrin-binding proteins TbpB and TbpA, both of which have been implied to be important virulence factors. In order to identify additional iron-regulated proteins, we cloned and analyzed the region upstream of the transferrinbinding protein genes in an A. pleuropneumoniae serotype 7 strain. We located immediately upstream of the tbpB gene two open reading frames which were 43% homologous to the neisserial ExbBD protein genes. By raising specific antibodies, we showed that ExbB is expressed under iron-limiting growth conditions only, and RT-PCR analysis revealed that the exbBD genes and the tbpB gene are transcribed on a single polycistronic mRNA. By constructing an isogenic and nonpolar exbBD mutant, we showed that the exbBD genes are required by A. pleuropneumoniae for utilization of transferrin-bound iron. Using PCR and Western blotting, we showed that the genetic organization found in A. pleuropneumoniae serotype 7 is similar in all 12 A. pleuropneumoniae serotype reference strains.

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Section: Methodsmentioning

confidence: 99%

Actinobacillus pleuropneumoniaeIron Transport: a Set ofexbBDGenes Is Transcriptionally Linked to thetbpBGene and Required for Utilization of Transferrin-Bound Iron

et al. 2000

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“…All bacteria were grown with aeration at 37 "C in the appropriate medium rendered iron-restricted by the addition of 100 pM ethylenediamine di-o-hydroxyphenylacetic acid (EDDA). Haemophilus strains were grown as described by Williams & Brown (1986), in brain heart infusion (BHI) broth supplemented with 2 pg PNAD ml-' and 0.5 pg protoporphyrin I X ml-' ; A. pleuropneumoniae strains were grown in a medium based on that described by Niven et al (1989), which consists of 2 YO (w/v) tryptone, 0.5 YO (w/v) yeast extract, 100 mM NaC1, 10 mM KC1, 10 mM Na,HPO,, 10 mM glucose, PNAD to a final concentration of 5 pM and the pH adjusted to 7.4 with KOH. N .…”

Section: Introductionmentioning

confidence: 99%

Conservation and antigenic cross-reactivity of the transferrin-binding proteins of Haemophilus influenzae, Actinobacillus pleuropneumoniae and Neisseria meningitidis

et al. 1996

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Haemophilus influenzae acquires iron from the iron-transporting glycoprotein transferrin via a receptor-mediated process. This involves two outer-membrane transferrin-binding proteins (Tbps) termed T b p l and Tbp2 which show considerable preference for the human form of transferrin. Since the Tbps are attracting considerable attention as potential vaccine components, we used transferrin affinity chromatography to examine their conservation amongst 28 H. influenzae type b strains belonging to different outer-membrane-protein subtypes as well as six non-typable strains. Whole cells of all type b and nontypable strains examined bound human transferrin; whilst most strains possessed a T b p l of approximately 105 kDa, the molecular mass of Tbp2 varied from 79 to 94 kDa. Antisera raised against affinity-purified native H. influenzae TbpllTbp2 receptor complex cross-reacted on Western blots with the respective Tbps of all the Haemophilus strains examined. When used to probe Neisseria meningitidis Tbps, sera from each of four mice immunized with the Haemophilus TbplR complex recognized the 68 kDa Tbp2 of N. meningitidis strain B16B6 but not the 78 kDa Tbp2 of N. meningitidis strain 70942. Serum from one mouse also reacted weakly with T b p l of strain B16B6. Apart from a weak reaction with the Tbp2 of a serotype 5 strain, this mouse antiserum failed to recognize the Tbps of the porcine pathogen A. pleuropneumoniae. However, a monospecif ic polyclonal antiserum raised against the denatured Tbp2 of Neisseria meningitidis B16B6 recognized the Tbps of all Haemophilus and Actinobacillus strains examined. Since H. influenzae forms part of the natural flora of the upper respiratory tract, human sera were screened for the presence of antibodies to the Tbps. Sera from healthy adults contained antibodies which recognized both T b p l and Tbp2 from H. influenzae but not N. meningitidis. Convalescent sera from meningococcal meningitis patients contained antibodies which, on Western blots, recognized the Tbps2s of both pathogens. These data demonstrate the existence of shared epitopes on the Tbps of H. influenre, N. meningitidis and A. pleuropneumoniae despite their transferrin species specificity.

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“…This bacterium can use hemoglobin, heme-containing compounds (2), porcine transferrin (15), and exogenously supplied siderophores (6) as sources of iron. Under iron-limiting conditions, A. pleuropneumoniae expresses two transferrin binding proteins, TbpA (ϳ100 kDa) and TbpB (ϳ60 kDa) (10,11), both of which have been documented as virulence factors (1).…”

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confidence: 99%

fhuAofActinobacillus pleuropneumoniaeEncodes a Ferrichrome Receptor but Is Not Regulated by Iron

et al. 2003

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The swine pathogen Actinobacillus pleuropneumoniae possesses a 75-kDa outer membrane protein (OMP), FhuA, the receptor for ferrichrome, a hydroxamate-type siderophore. Polyclonal serum to FhuA reacted with OMP preparations from 12 serotypes of A. pleuropneumoniae under conditions of iron repletion and restriction. Reverse transcription-PCR confirmed that A. pleuropneumoniae fhuA expression is not upregulated in response to low iron levels. An A. pleuropneumoniae fhuA deletion mutant was generated and showed abolishment of ferrichrome uptake.

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Responses of Haemophilus pleuropneumoniae to iron restriction: changes in the outer membrane protein profile and the removal of iron from porcine transferrin

Cited by 52 publications

References 42 publications

Actinobacillus pleuropneumoniaeIron Transport: a Set ofexbBDGenes Is Transcriptionally Linked to thetbpBGene and Required for Utilization of Transferrin-Bound Iron

Actinobacillus pleuropneumoniaeIron Transport: a Set ofexbBDGenes Is Transcriptionally Linked to thetbpBGene and Required for Utilization of Transferrin-Bound Iron

Conservation and antigenic cross-reactivity of the transferrin-binding proteins of Haemophilus influenzae, Actinobacillus pleuropneumoniae and Neisseria meningitidis

fhuAofActinobacillus pleuropneumoniaeEncodes a Ferrichrome Receptor but Is Not Regulated by Iron

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