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J de Graaff

Publications and source records attributed to J de Graaff.

At least 19 recordsLinked to original sources
PubMed1999

The inflammatory response in CD1 mice shortly after infection with a CagA+/VacA+ Helicobacter pylori strain.

To investigate the early events of Helicobacter pylori infection in a mouse model, CD1 mice were infected with a type I (CagA+/VacA+) H. pylori strain. Up to 4 weeks after infection the majority of gastric tissue biopsies were positive in culture. Immunohistochemical analysis showed that inflammatory changes started to occur after 3 weeks. Four weeks after infection a significant increase in T cells was observed in the cardia/corpus region of the stomachs of infected mice. These T cells were CD4+ and CD8+, and they were located in an area with increased expression of MHC class II antigens. In 50% of the infected mice also an increased number of mast cells was seen. Furthermore, aggregates of B and T cells were present in the submucosa. Characterization of cytokines by immunohistochemistry showed an increase in IL-5-secreting cells in the inflamed area of the infected stomach. No difference was observed between interferon-gamma (IFN-gamma)-, IL-4- and IL-10-secreting cells in control and infected mice. These results suggest that no polarized T-helper cell response was present at this early phase of infection. Infection with H. pylori also induced a serum response and especially IgG was increased after 4 weeks of infection. However, no particular increase in IgG1, IgG2a or IgG3 isotype was observed. Part of the serum antibodies was directed against lipopolysaccharide (LPS), but no evidence for anti-Lewis antibodies or antibodies against epitopes on the gastric mucosa was found.

Animals↗
PubMed1999

Cloning of fibA, encoding an immunogenic subunit of the fibril-like surface structure of Peptostreptococcus micros.

Although we are currently unaware of its biological function, the fibril-like surface structure is a prominent characteristic of the rough (Rg) genotype of the gram-positive periodontal pathogen Peptostreptococcus micros. The smooth (Sm) type of this species as well as the smooth variant of the Rg type (RgSm) lack these structures on their surface. A fibril-specific serum, as determined by immunogold electron microscopy, was obtained through adsorption of a rabbit anti-Rg type serum with excess bacteria of the RgSm type. This serum recognized a 42-kDa protein, which was subjected to N-terminal sequencing. Both clones of a lambdaTriplEx expression library that were selected by immunoscreening with the fibril-specific serum contained an open reading frame, designated fibA, encoding a 393-amino-acid protein (FibA). The 15-residue N-terminal amino acid sequence of the 42-kDa antigen was present at positions 39 to 53 in FibA; from this we conclude that the mature FibA protein contains 355 amino acids, resulting in a predicted molecular mass of 41,368 Da. The putative 38-residue signal sequence of FibA strongly resembles other gram-positive secretion signal sequences. The C termini of FibA and two open reading frames directly upstream and downstream of fibA exhibited significant sequence homology to the C termini of a group of secreted and surface-located proteins of other gram-positive cocci that are all presumably involved in anchoring of the protein to carbohydrate structures. We conclude that FibA is a secreted and surface-located protein and as such is part of the fibril-like structures.

Amino Acid Sequence↗
PubMed1998

Genomic DNA fingerprinting of clinical isolates of Helicobacter pylori by REP-PCR and restriction fragment end-labelling.

Genetic diversity of 32 Helicobacter pylori strains isolated from patients with gastritis, gastric or duodenal ulcer, carcinoma, or lymphoma was determined by repetitive sequence element polymerase chain reaction (REP-PCR), and by the new typing method restriction fragment end-labelling (RFEL). Furthermore, these two methods were used to investigate a possible correlation between clinical symptoms and the genetic background of Helicobacter pylori. Both REP-PCR and RFEL revealed 31 different patterns for the 32 strains tested, but the pair of isolates with identical REP-PCR patterns was not the same as the pair of isolates with identical RFEL patterns. Computer-assisted analysis of the DNA fingerprints was used to determine similarity coefficients. This analysis revealed no clustering of disease-specific strains by any of the two methods.

DNA Fingerprinting↗
PubMed1998

Local cellular immune response in the acute phase of gastritis in mice induced chemically and by Helicobacter pylori.

Gastritis was induced in mice by oral administration of acetic acid 5%, a cagA positive Helicobacter pylori strain, or both. The induction of a mild gastritis by acetic acid before inoculation with H. pylori resulted in a slight but not significantly decreased colonisation rate. To study the initial stage of inflammation, the presence of gastric lymphoid and non-lymphoid cells was studied by immunohistochemistry during the first 2 weeks after induction of gastritis. Treatment with acetic acid alone or in combination with H. pylori resulted in an increase in the number of neutrophils in the mucosa and submucosa, without evident epithelial damage. The influx of neutrophils was most prominent in the mice that received a combined treatment of acetic acid and H. pylori. Macrophages were also increased in both acetic acid and acetic acid plus H. pylori-treated groups, although a different kinetic pattern was present in these groups. In mice infected with H. pylori alone, only a slight but not significant increase in neutrophils and macrophages was observed. The early presence of lymphoid aggregates in the gastric mucosa of mice in which colonisation was shown with H. pylori was remarkable. This phenomenon was not seen in control mice, in mice that received acetic acid alone or when colonisation was not shown. These data suggest that gastritis induced by a chemical agent such as acetic acid occurs by a different mechanism than gastritis induced by H. pylori and that the continued presence of H. pylori is required for local lymphocyte activation.

Acetic Acid↗
PubMed1998

Pathogenicity of Peptostreptococcus micros morphotypes and Prevotella species in pure and mixed culture.

Recently, an atypical rough colony morphotype of Peptostreptococcus micros, a species which is found in ulcerating infections, including periodontitis, was isolated. The virulence of morphotypes alone and in combination with Prevotella intermedia and P. nigrescens was investigated both in vivo and in vitro. All strains tested induced abscesses containing fluid pus in a mouse skin model, and lesions caused by monocultures of the rough morphotype strains of P. micros were statistically significantly larger than those induced by the smooth morphotype strains. Inocula containing both morphotypes produced similar sized abscesses compared to mono-inocula containing the same bacterial load. Both Prevotella species induced small abscesses when inoculated alone, and when Pr. nigrescens was inoculated with one of the other strains, the abscesses were not significantly different from the abscesses induced by the mono-infections of this strain. Synergy, in terms of higher numbers of colony forming units (cfu) in the mixed inocula, was found for all combinations of the rough morphotypes of P. micros and both Prevotella spp. Pus from abscesses caused by combinations of Peptostreptococcus and Prevotella spp. transmitted the infection to other mice, but no abscesses were formed in mice inoculated with pus induced by mono-inocula. These results demonstrated synergic activity between both rough and smooth P. micros strains and oral Prevotella strains. The in-vitro co-culture experiments produced no evidence of growth stimulation. The effect of P. micros strains on the immune system was investigated by testing their ability to initiate luminol-dependent chemiluminescence of polymorphonuclear leucocytes in the presence and absence of human serum. In the latter, the rough morphotype strains initiated higher counts than the smooth morphotype strains. Further work is needed to elucidate the difference in virulence between the smooth and the rough morphotype cells of P. micros and the nature of the interaction with the Prevotella spp.

Abscess↗
PubMed1998

Colonization with mutans streptococci and lactobacilli and the caries experience of children after the age of five.

Some authors suggest that there is a "window of infectivity" for mutans streptococci (MS) at an early age, after which colonization is not likely to occur. Moreover, children with low or non-detectable levels of MS at an early age are at lower risk for caries. It was our aim to study caries experience and colonization with MS and lactobacilli in a group of children at 11 years of age who had been documented to have been MS-free until 5 years of age. For this group, the mfs and MFS values at 11 years of age were found to be 1.12+/-2.81 and 0.44+/-0.88, respectively. These values are much lower than those of a control group of 11-year-old children who had always been MS-positive since the age of 2, being 3.10+/-3.43 and 1.20+/-1.91, respectively. Of the 30 children without detectable MS up to the age of 5, 22 had acquired MS at 11 years of age, but their MS counts were significantly lower than those of the control group. No correlation was observed between the levels of MS of the children and the mothers. In children at 11 years of age, no differences were found in the numbers of lactobacilli and sugar intakes per week between the children MS-free and the children MS-colonized at 5 years of age. The present study indicates that the acquisition of MS is still possible after the so-called "window of infectivity" and that a delayed acquisition of MS may reduce the caries experience in the primary and permanent dentition at a later age.

Age Factors↗
PubMed1998

Transmission of mutans streptococci between mothers and children with cleft lip and/or palate.

OBJECTIVE: The aim of this study was to investigate the transmission of Streptococcus mutans between children with cleft lip and/or palate and their mothers. DESIGN: Saliva samples of 21 mother-child pairs were collected and cultured on plates containing a selective growth medium for mutans streptocci. At least five separate colonies of each colony morphotype were isolated. A polymerase chain reaction (PCR) with randomly chosen primers was used to type the isolates. RESULTS: The number of morphotypes and PCR types was significantly lower in the children than in the mothers. Significant correlations were found between the number of morphotypes and PCR types, in the children as well as in the mothers. In only 38% of the mother-child pairs were the same PCR types found in mother and child. CONCLUSIONS: This suggests that S. mutans had been transmitted from mother to child in one-third of the population studied. No correlations were found among the number of colony-forming units, the number of colony-colony-morphotypes, and the number of PCR types of the mothers and transmission. Similar PCR types in mother and child were found significantly more often in children who had more than one PCR type. The results indicate that transmission of S. mutans from mother to child is not frequent in children with oral cleft. This may have consequences for preventive treatment of cleft lip and/or palate children and their mothers.

Chi-Square Distribution↗
PubMed1997

Endopeptidase activities of selected Porphyromonas spp., Prevotella spp. and Fusobacterium spp. of oral and non-oral origin.

The ability of three Porphyromonas spp., seven Prevotella spp., seven Fusobacterium spp. and two related Bacteroides spp. (B. levii and B. macacae) to degrade an extensive range of synthetic endo-, amino- and diamino peptidase substrates linked to the fluorescent leaving group 7-amido-4-methylcoumarin (NHMec) was investigated. Many more species than was previously recognized exhibited peptidase activities, albeit at lower levels than those already described for Porphyromonas gingivalis. Detection of chymotrypsin-like activity was dependent on which of three NHMec-linked substrates was used, but all species exhibited degradative activity with at least one of these substrates. Elastase-like activity was detected in all species though not all species reacted with each of the elastase substrates. Glycylprolyl peptidase activity was detected in all of the species tested with the exception of F. mortiferum, F. gonidiaformans, F. naviforme and F. necrophorum. While the detection of peptidase activities does not appear to be useful for the differentiation of species within the genera Bacteroides and Prevotella, its ability to differentiate species of the genus Porphyromonas or Fusobacterium warrants further investigation.

Alanine↗
PubMed1997

Characterization of a periplasmic protein involved in iron utilization of Actinobacillus actinomycetemcomitans.

The periodontopathic bacterium Actinobacillus actinomycetemcomitans possesses a 35-kDa periplasmic iron-repressible protein. Its regulation is mediated by the Fur protein, as was inferred from the Fur-binding consensus sequence at the -35 position of the gene for the 35-kDa protein and from the relaxed expression of the gene in a mutant with an altered Fur-binding sequence. The 35-kDa protein, designated AfuA, has strong homology to HitA and FbpA of Haemophilus influenzae and Neisseria meningitidis, respectively, which serve as periplasmic iron transport proteins.

Aggregatibacter actinomycetemcomitans↗
PubMed1997

Effect of antibodies on the chain length and growth of Streptococcus sobrinus.

Passive immunization has been suggested as a method to prevent colonization of teeth by mutans streptococci. However, the mechanism of action of antibodies, both polyclonal and monoclonal, is not clear. In this study we investigated the effect of polyclonal antibodies (pAbs) and a monoclonal antibody (OMVU10) on the chain length and growth of Streptococcus sobrinus. During growth in broth S. sobrinus formed significantly longer chains in the presence of pAbs in comparison to pre-immune serum (p < 0.01), but pAbs did not influence the growth rate of S. sobrinus in broth. OMVU10 did not influence the growth rate nor the chain length. In order to study the effect of the antibodies on adhesion and growth on a surface, S. sobrinus was grown on hydroxyapatite discs in the presence of two other bacteria, Streptococcus oralis and Actinomyces viscosus. No effect of the antibodies was found on the total cultivable count of the three bacteria after growth on hydroxyapatite discs. However, the morphology of the S. sobrinus microcolonies was different after growth in the presence of both pAbs and OMVU10. The colonies were less dense and chains of S. sobrinus could be seen using a confocal laser microscope. After growth in the presence of the control antibodies, the colonies were dense and no long chains could be observed. It was concluded that pAbs influenced the chain length, in broth and on hydroxyapatite discs, and the colony morphology of S. sobrinus on hydroxyapatite discs, whereas OMVU10 influenced the colony morphology of S. sobrinus on hydroxyapatite discs.

Aggregatibacter actinomycetemcomitans↗
PubMed1996

Human serum antibody response against iron-repressible outer membrane proteins of Helicobacter pylori.

In Helicobacter pylori, in vitro iron limitation induces the expression of several iron repressible outer membrane proteins (IROMPs), which are not expressed under normal growth conditions. To substantiate their proposed role in virulence of H. pylori, we determined whether these IROMPs are also expressed in vivo. Therefore, we tested whether sera of patients with H. pylori infection contained antibodies against IROMPs. All sera from 20 H. pylori positive patients showed a clear immune response against a 77 kDa heme-binding IROMP in an immunoblot assay. Antibody responses against the other IROMPs were also found, but with lower frequencies. Serum samples from 18 patients negative for H. pylori infection did not show any immunoreactivity with IROMPs. These results indicate that the IROMPs of H. pylori are immunogenic and are expressed in vivo.

Antibodies, Bacterial↗
PubMed1996

Effect of antibodies on chemiluminescence and on killing of Streptococcus sobrinus by polymorphonuclear leukocytes.

The effect of a polyclonal antiserum and OMVU10, a monoclonal antibody reactive with Antigen B of Streptococcus sobrinus, on the interaction of polymorphonuclear leukocytes with S. sobrinus was studied, using chemiluminescence and bacterial killing assays. Increased stimulation of neutrophils as measured in the chemiluminescence assays was established when S. sobrinus was preincubated with polyclonal antiserum or when polyclonal antiserum was added to the reaction mixture. Higher counts were measured in comparison to preimmune serum. After 90 min, 52% of S. sobrinus preincubated with polyclonal antiserum was killed. Killing was also increased when polyclonal antiserum was added to the reaction mixture in comparison to the controls. No killing was found when bacteria were preincubated with OMVU10 or when OMVU10 was added to the reaction mixture in comparison to Clone 24, a control antibody.

Antibodies, Bacterial↗
PubMed1996

Discrepancy between culture and DNA probe analysis for the detection of periodontal bacteria.

The purpose of this study was to compare a commercially available DNA probe technique with conventional cultural techniques for the detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in subgingival plaque samples. Samples from 20 patients with moderate to severe periodontitis were evaluated at baseline and during a 15 months period of periodontal treatment. Paperpoints from 4 periodontal pockets per patient were forwarded to Omnigene for DNA probe analysis, and simultaneously inserted paperpoints from the same pockets were analyzed by standard culture techniques. In addition, mixed bacterial samples were constructed harbouring known proportions of 25 strains of A. actinomycetemcomitans, P. gingivalis and P. intermedia each. A relatively low concordance was found between both methods. At baseline a higher detection frequency was found for A. actinomycetemcomitans and P. gingivalis for the DNA probe technique; for P. intermedia the detection frequency by culture was higher. For A. actinomycetemcomitans, 21% of the culture positive samples was positive with the DNA probe. Testing the constructed bacterial samples with the DNA probe method resulted in about 16% false positive results for the 3 species tested. Furthermore, 40% of P. gingivalis strains were not detected by the DNA probe. The present data suggest that at least part of the discrepancies found between the DNA probe technique used and cultural methods are caused by false positive and false negative DNA probe results. Therefore, the value of this DNA probe method for the detection of periodontal pathogens is questionable.

Adult↗
PubMed1996

Evaluation of the long-term efficacy and safety of locally-applied minocycline in adult periodontitis patients.

The objectives of the present study were to establish in a long-term investigation the safety as well as the clinical and microbiological efficacy of scaling and rootplaning combined with local application of 2% minocycline hydrochloride-gel versus placebo-gel in patients with moderate to severe chronic adult periodontitis. This was an 18 months, randomized, double-blind, parallel, comparative study, in which 20 healthy patients with moderate to severe chronic periodontitis participated. At baseline, all patients received professional oral hygiene-instruction and supra- and subgingival scaling and root planing. The minocycline-gel was applied subgingivally baseline, 2 weeks, 1, 3, 6, 9 and 12 months. Microbiological evaluation was carried out using DMDx to identify the following bacteria: Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, Campylobacter rectus, Fusobacterium nucleatum and Treponema denticola. In addition standard microbiological techniques were used for the detection of P. gingivalis, P. intermedia, P. micros, A. actinomycetemcomitans, C. rectus, F. nucleatum, C. albicans and Enterobacteriaceae. Results showed a statistically significant improvement for all clinical parameters irrespective of the treatment modality. No differences were observed between test and control with regard to probing depth and attachment level. The DMDx data showed a significant reduction in both the numbers and the prevalence over the 15 months period, but no significant difference between groups. Culture data showed that at baseline two-third were positive for P. gingivalis and P. intermedia. Analysis over the 18 month period showed no significant difference between the two treatment modalities. C. albicans and Enterobacteriaceae were detected only in small proportions at each time interval in a limited number of patients. No adverse reactions were observed during the trial period. The present patient group responded favourably to scaling and rootplaning, but did not benefit from an effect of local of minocycline. Subgingival debridement in combination with oral hygiene instruction by itself has been shown to be effective. It remains to be studied whether local application of minocycline can be effective as an adjunct to mechanical therapy in sites that respond poorly to conventional treatment.

Administration, Topical↗
PubMed1996

Typing of Helicobacter pylori with monoclonal antibodies against Lewis antigens in lipopolysaccharide.

Recently, it has been shown that the lipopolysaccharide (LPS) O antigen of Helicobacter pylori contains Lewis x (Lex), Lewis y (Ley), or both Lex and Ley antigens. We applied a serotyping method for H. pylori by an enzyme-linked immunosorbent assay with monoclonal antibodies (MAbs) specific for these antigens and the related fucosylated H type 1 (H1) antigen. The selected MAbs recognized the Lex and/or Ley structures in the LPS of H. pylori. The agreement between the results of biochemical compositional analysis and the serological data validated our serotyping system. A total of 152 strains from different geographic origins (The Netherlands, Canada, Poland, Italy, and People's Republic of China) were examined for typeability based on the presence of Lewis antigens. One hundred twenty-nine (84.9%) strains were typeable, and 12 different serotyping patterns were observed; 80.9% of the strains contained Lex and/or Le(y) antigens, and 18.4% reacted with the MAb against the related H1 antigen either alone or in combination with the Lex and/or Ley antigen. Our results show that the Lex and Ley antigens are frequently encountered in the LPS of H. pylori strains from various geographic origins. This typing method is an easy-to-perform technique, which can be used for strain differentiation in epidemiological studies of H. pylori infections.

Antibodies, Bacterial↗
PubMed1996

Practical diagnosis of Helicobacter pylori infection.

When treatment of Helicobacter pylori infection is considered, a reliable diagnosis is essential. Different biopsy-based invasive diagnostic tests are available for diagnosis in the individual patient. Non-invasive tests are of value in epidemiological studies, and have a role in the follow-up of treatment. In this review the sensitivity, specificity and appropriateness of all available tests for the diagnosis of infection in the individual patient are discussed.

Breath Tests↗
PubMed1995

Genomic DNA fingerprinting by restriction fragment end labeling.

A typing method for bacteria was developed and applied to several species, including Escherichia coli and Actinobacillus actinomycetemcomitans. Total genomic DNA was digested with a restriction endonuclease, and fragments were enabled with [alpha-32P]dATP by using the Klenow fragment of DNA polymerase and separated by electrophoresis in 6% polyacrylamide/8 M urea (sequencing gel). Depending on the restriction endonuclease and the bacterium, the method produced approximately 30-50 well-separated fragments in the size range of 100-400 nucleotides. For A. actinomycetemcomitans, all strains had bands in common. Nevertheless, many polymorphisms could be observed, and the 31 strains tested could be classified into 29 distinct types. Furthermore, serotype-specific fragments could be assigned for the three serotypes investigated. The method described is very sensitive, allowing more distinct types to be distinguished than other commonly used typing methods. When the method was applied to 10 other clinically relevant bacterial species, both species-specific bands and strain-specific bands were found. Isolates from different locations of one patient showed indistinguishable patterns. Computer-assisted analysis of the DNA fingerprints allowed the determination of similarity coefficients. It is concluded that genomic fingerprinting by restriction fragment end labeling (RFEL) is a powerful and generally applicable technique to type bacterial species.

DNA Fingerprinting↗