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At least 19 recordsLinked to original sources
PubMed2006

Reclassification of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, Haemophilus paraphrophilus and Haemophilus segnis as Aggregatibacter actinomycetemcomitans gen. nov., comb. nov., Aggregatibacter aphrophilus comb. nov. and Aggregatibacter segnis comb. nov., and emended description of Aggregatibacter aphrophilus to include V factor-dependent and V factor-independent isolates.

The aim of this study was to reinvestigate the relationships and the generic affiliations of the species Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus, Haemophilus paraphrophilus and Haemophilus segnis. The nicotinamide phosphoribosyltransferase gene (nadV) conferring V factor-independent growth was identified in Haemophilus aphrophilus. The gene encodes a polypeptide of 462 amino acids that shows 74.5 % amino acid sequence identity to the corresponding enzyme from Actinobacillus actinomycetemcomitans. Ten isolates of Haemophilus paraphrophilus all carried a nadV pseudogene. DNA from Haemophilus aphrophilus was able to transform Haemophilus paraphrophilus into the NAD-independent phenotype. The transformants carried a full-length nadV inserted in the former locus of the pseudogene. The DNA-DNA relatedness between the type strains of Haemophilus aphrophilus and Haemophilus paraphrophilus was 77 %. We conclude that the division into two species Haemophilus aphrophilus and Haemophilus paraphrophilus is not justified and that Haemophilus paraphrophilus should be considered a later heterotypic synonym of Haemophilus aphrophilus. Forty strains of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus and Haemophilus segnis were investigated by multilocus sequence analysis. The 40 strains form a monophyletic group clearly separate from other evolutionary lineages of the family Pasteurellaceae. We propose the transfer of Actinobacillus actinomycetemcomitans, Haemophilus aphrophilus and Haemophilus segnis to a new genus Aggregatibacter gen. nov. as Aggregatibacter actinomycetemcomitans comb. nov. (the type species; type strain ATCC 33384(T)=CCUG 13227(T)=CIP 52.106(T)=DSM 8324(T)=NCTC 9710(T)), Aggregatibacter aphrophilus comb. nov. (type strain ATCC 33389(T)=CCUG 3715(T)=CIP 70.73(T)=NCTC 5906(T)) and Aggregatibacter segnis comb. nov. (type strain HK316(T)=ATCC 33393(T)=CCUG 10787(T)=CCUG 12838(T)=CIP 103292(T)=NCTC 10977(T)). The species of the genus Aggregatibacter are independent of X factor and variably dependent on V factor for growth in vitro.

Actinobacillus↗
PubMed2006

TdeA, a TolC-like protein required for toxin and drug export in Aggregatibacter (Actinobacillus) actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans is an oral bacterium that causes localized aggressive periodontitis (LAP) and extra-oral infections such as sub-acute infective endocarditis. As part of its array of virulence factors, A. actinomycetemcomitans produces leukotoxin (LtxA), a member of the RTX family of toxins. LtxA kills human leukocytes and we have recently shown that the toxin is required for beta-hemolysis by A. actinomycetemcomitans on solid medium. In other RTX toxin-producing bacteria, an outer membrane channel-forming protein, TolC, is required for toxin secretion and drug export. We have identified an ORF in A. actinomycetemcomitans that encodes a putative protein having predicted structural properties similar to TolC. Inactivation of this ORF resulted in a mutant that was no longer beta-hemolytic and did not secrete LtxA. This mutant was significantly more sensitive to antimicrobial agents compared to the wild type strain and was unable to export the antimicrobial agent berberine. Thus, this ORF was named tdeA for "toxin and drug export". Examination of the DNA sequence surrounding tdeA revealed two upstream ORFs that encode proteins similar to the drug efflux proteins, MacA and MacB. Inactivation of macB in A. actinomycetemcomitans did not alter the drug sensitivity profile or the hemolytic activity of the mutant. The genes macA, macB and tdeA are organized as an operon and are constitutively expressed as a single transcript. These results show that A. actinomycetemcomitans indeed requires a TolC-like protein for LtxA secretion and that this protein, TdeA, also functions as part of a drug efflux system.

Aggregatibacter actinomycetemcomitans↗
PubMed2006

Regulation of Aggregatibacter (Actinobacillus) actinomycetemcomitans leukotoxin secretion by iron.

The gram-negative oral and systemic pathogen Aggregatibacter (Actinobacillus) actinomycetemcomitans produces a leukotoxin (LtxA) that is a member of the RTX (repeats in toxin) family of secreted bacterial toxins. We have recently shown that LtxA has the ability to lyse erythrocytes, which results in a beta-hemolytic phenotype on Columbia blood agar. To determine if LtxA is regulated by iron, we examined beta-hemolysis under iron-rich and iron-limiting conditions. Beta-hemolysis was suppressed in the presence of FeCl3. In contrast, strong beta-hemolysis occurred in the presence of the iron chelator deferoxamine. We found that secretion of LtxA was completely inhibited by free iron, but expression of ltxA was not regulated by iron. Free chromium, cobalt, and magnesium did not affect LtxA secretion. Other LtxA-associated genes were not regulated by iron. Thus, iron appears to play an important role in the regulation of LtxA secretion in A. actinomycetemcomitans in a manner independent of gene regulation.

Aggregatibacter actinomycetemcomitans↗
PubMed2005

16S rDNA PCR-denaturing gradient gel electrophoresis in determining proportions of coexisting Actinobacillus actinomycetemcomitans strains.

Certain serotypes of Actinobacillus actinomycetemcomitans seem to prefer coexistence in vivo. The 16S rDNA PCR-denaturing gradient gel electrophoresis (DGGE) was tested for its capability to distinguish coexisting A. actinomycetemcomitans strains of different serotypes or genetic lineages and to determine their proportions in vitro. The migration pattern of the PCR amplicon from serotype c differed from those of the other serotypes. Contrary to the strains of serotypes c, d, and e, strains of serotypes a, b, and f consistently demonstrated intra-serotype migration patterns similar to each other. Since the migration patterns differed between serotype c and b strains a strain of each was used to determine their proportional representation in a strain mixture. The strains were distinguishable from each other above the 5% PCR-DGGE detection level (12.5 ng DNA/1.5 x 10(6) cells). DGGE provides a promising tool for in vitro studies on the coexistence of different genetic lineages of A. actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans↗
PubMed1993

Identification of Actinobacillus actinomycetemcomitans by leukotoxin gene-specific hybridization and polymerase chain reaction assays.

Eleven strains of Actinobacillus actinomycetemcomitans isolated from cases of systemic infections, local abscesses, and periodontitis were identified by genetic assays using the leukotoxin gene as the target. We have developed a polymerase chain reaction (PCR) assay, based on the leukotoxin structural gene of this pathogen, which clearly identified all tested strains of A. actinomycetemcomitans and separated them from the closely related Haemophilus aphrophilus as well as other bacterial species. Furthermore, DNA-DNA hybridization was performed with the cloned partial leukotoxin structural gene (lktA) as a probe, which again clearly distinguished A. actinomycetemcomitans from H. aphrophilus, parts of the normal oral flora, and species harboring RTX (repeats in toxin) family-related cytotoxins. The PCR fragment amplified from the leukotoxin structural gene gave results similar to those given by the cloned leukotoxin gene when used as a probe in hybridization experiments. The hybridization and PCR assays described here are fundamental improvements for the identification of A. actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans↗
PubMed1990

[Analysis of antigens of Actinobacillus actinomycetemcomitans with monoclonal antibodies].

The purpose of this study was to analyze antigens of Actinobacillus actinomycetemcomitans. Fifteen hybridomas producing monoclonal antibodies (MAbs) against A. actinomycetemcomitans strain Y4 were obtained. These hybridomas were divided into three groups (Group 1, Group 2 and Group 3) on their MAbs' specificity. The MAbs (MAb S1-S8) produced by Group 1 hybridomas reacted with serotype b-specific antigen of A. actinomycetemcomitans. The MAbs (MAb L1-L3) produced by Group 2 hybridomas reacted with lipopolysaccharides (LPSs) of all serotypes of A. actinomycetemcomitans. The high-molecular-weight peak (peak A) and the low-molecular-weight peak (peak B) were separated by gel-filtration of the phenol-water extract (PWE) of strain Y4. MAb S5 reacted with peak A, and MAb L2 reacted with peak B. Peak B bound to a polymyxin affinity column, but peak A did not. These findings indicate that peak A was serotype-specific antigen and peak B was LPS. MAbs P1, P2, P3 and P4 produced by Group 3 hybridomas reacted with 81 kDa, 64 kDa, 64 kDa and 40 kDa protein antigens, respectively. Preincubation of strain Y4 whole cells with MAb P3 inhibited significantly the adherence of the cells to human buccal epithelium cells (HBECs). These findings suggest that the 64 kDa protein antigen might participate in the adherence of A. actinomycetemcomitans to HBECs.

Aggregatibacter actinomycetemcomitans↗
PubMed1995

Inducible bacteriophages of Actinobacillus actinomycetemcomitans.

Doses of 0.1 to 1.0 micrograms/ml of mitomycin C induced cell lysis of six of eight strains of Actinobacillus actinomycetemcomitans tested. Infectious phages were induced from ATCC strains 43717, 29524, 33384, and 43719; non-plaque-forming, possibly defective phages were induced from ATCC strains 29522 and 29523. No phages were detected in strain FDC 651 or ATCC strain 43718. No correlation between lysogeny and leukotoxin production or serotype of the strains could be established. Gel electrophoresis of phage DNAs indicated that the induced phages were of three types, based on size. By electron microscopy, the phages were found to belong to either morphotype A1 or morphotype B1; no other morphotypes were observed. Curing experiments led to the isolation of nonlysogenic derivatives of two strains, which supported plaque formation by the phages they originally carried. On the basis of our results, lysogeny appears to be widespread in A. actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans↗
PubMed1994

Characterization of plasmid pVT745 isolated from Actinobacillus actinomycetemcomitans.

A 25.1-kb plasmid, pVT745, was isolated from Actinobacillus actinomycetemcomitans strain VT745. This plasmid hybridized under stringent conditions to chromosomal DNA from 15 A. actinomycetemcomitans isolates obtained from geographically diverse regions of the United States. Southern blot analyses of HincII-digested A. actinomycetemcomitans genomic DNA revealed five strain-specific patterns of hybridization, which clearly indicated that intact pVT745 was not inserted into any of the genomes. Plasmid pVT745 was digested with BamHI and PstI into three, non-overlapping fragments of approximately 7.0, 8.0, and 10.0 kb. The fragments were cloned in Escherichia coli on the low copy number vector pGB2. Although these three fragments exhibited no cross-hybridization, genomic DNA from isolates representing each of the strain-specific patterns hybridized with two or three of the cloned fragments. The results suggest that two or more unique sequences within pVT745 also may be present in genomic DNA obtained from various strains of A. actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans↗
PubMedDate not supplied

Stress response in Actinobacillus actinomycetemcomitans: induction of general and specific stress proteins.

Stress proteins are highly conserved proteins that are essential for cell survival. In this study, the induction of general and specific stress proteins in Actinobacillus actinomycetemcomitans cells subjected to different stress conditions was evaluated by two-dimensional SDS-PAGE analysis. Twenty-eight (up- or down)regulated proteins, including DnaK and GroEL proteins, were identified as general stress proteins. In addition, eighteen regulated proteins were classified as pH stress-specific proteins, ten as acid stress-specific proteins, five as alkaline stress-specific proteins, three as heat stress-specific proteins, and ten as acid/heat stress-specific proteins. Further proteomic studies are required to determine the exact nature of the proteins regulated during the stress response of A. actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans↗
PubMed1999

Genotyping of Actinobacillus actinomycetemcomitans serotype d isolates based on polymerase chain reaction.

The aims of the study were to determine the genetic diversity of the rare Actinobacillus actinomycetemcomitans serotype d and to compare the ability of the repetitive extragenic palindromic element (REP)-based polymerase chain reaction (PCR) with that of the arbitrarily primed (AP)-PCR to discriminate between and within A. actinomycetemcomitans serotypes. The material included 26 A. actinomycetemcomitans serotype d isolates, 3 reference strains, and 21 A. actinomycetemcomitans isolates, representing the previously described 17 AP-PCR genotypes from 4 serotypes (a, b, c and e). Among A. actinomycetemcomitans serotype d isolates (n = 26), the AP-PCR primer distinguished 2 genotypes, whereas the REP-primer pair (REP1R-I and REP2-I) and the (GACA)4 primer each produced one genotype. Among the total of 50 A. actinomycetemcomitans isolates, REP-primer pair distinguished 6 genotypes, the primer (GACA)4 7 genotypes, and the AP-PCR 19 genotypes. Among A. actinomycetemcomitans serotype a isolates (n = 6), REP-primer pair yielded 3 genotypes and (GACA)4 and AP-PCR primer 4 genotypes, and among serotype e isolates (n = 6) 3 genotypes. All serotype b isolates (n = 7), representing the AP-PCR genotypes 2, 9, 8, 12, 13, 16 and serotype c isolates (n = 5), AP-PCR genotypes 3, 4, 14, 15, belonged to the (REP1R-I and REP2-I)-PCR genotype 4 and to the (GACA)4-PCR genotype 4. In conclusion, based on both the AP-PCR method and the less discriminative REP-PCR methods, the present genotyping results indicated limited genetic diversity among serotype d isolates of A. actinomycetemcomitans.

Aggregatibacter actinomycetemcomitans↗
PubMed2000

Evidence for apoptosis of the majority of T cells activated in vitro with Actinobacillus actinomycetemcomitans.

Our previous studies had demonstrated that nearly half of all T cells stimulated with Actinobacillus actinomycetemcomitans are activated within a few hours. However, it was not known whether all of these T cells survive. The aim of the present study was to determine whether the T cells activated in response to A. actinomycetemcomitans undergo apoptosis. To that end, peripheral blood mononuclear cells were cultured at different time points in the presence of A. actinomycetemcomitans. Flow cytometric analysis demonstrated that, following exposure to a preparation of A. actinomycetemcomitans, T cells progressively externalized their plasma membrane phosphatidylserine, as measured by annexin V binding. Approximately half of all T cells bound annexin V by 96 h. During this period, Annexin V-positive T cells also incorporated propidium iodide suggesting loss of membrane integrity. The externalization of phosphatidylserine occurred at a higher rate among activated (CD69+) T cells, where roughly two-thirds became Annexin V-positive. Flow cytometric analysis also demonstrated shrinkage of the Annexin V-positive and propidium iodide-positive T cells. The data presented here provides evidence for the induction of apoptosis among the majority of the T cells responding to A. actinomycetemcomitans.

Adult↗