Protective Immunity to <i>Porphyromonas gingivalis</i> Infection in a Murine Model

Bird, P. S.; Gemmell, E.; Polak, B.; Paton, Robert G.; Sosroseno, Wihas; Seymour, G. J.

doi:10.1902/jop.1995.66.5.351

Cited by 47 publications

(44 citation statements)

References 25 publications

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“…BALB/c female mice (6-8 weeks old) were obtained from the University of Queensland Central Animal Breeding House. The immunization protocol has been described previously by Bird et al [25]. Twenty-nine mice were divided into five groups (six per group, with five mice in the control group).…”

Section: Immunization Proceduresmentioning

confidence: 99%

“…Detection of anti-F. nucleatum and P. gingivalis antibodies Serum samples were assayed for the presence of anti-F. nucleatum and P. gingivalis antibodies using an ELISA technique described by Bird et al [25,27,28]. Briefly, the protein concentrations of both organisms were determined (BCA Protein Assay Kit, Pierce Rockford, IL, USA) and the optimum coating concentration of F. nucleatum and P. gingivalis ascertained to be 5·0 mg/ml protein.…”

Section: Flow Cytometric Analysismentioning

confidence: 99%

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Effect of Fusobacterium nucleatum on the T and B cell responses to Porphyromonas gingivalis in a mouse model

Gemmell

Bird

Carter

et al. 2002

Clinical and Experimental Immunology

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Periodontal disease results from the inflammatory response to bacteria in dental plaque (reviewed in [1]). Although there are well over 300 different bacterial species in the plaque, progression to periodontitis depends not on bacterial load, but on the presence of specific periodontopathic bacteria. The major pathogens identified at the 1996 World Workshop in Periodontics as causative agents are Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Bacteroides forsythus [2]. P. gingivalis has been found in 15% of subjects in an Australian population, the prevalence increasing with increasing pocket depth [3]. In another study, Slots and Ting [4] found that 40-100% of adult periodontitis patients were positive for P. gingivalis. This high occurrence, together with the pathogenic potential of P. gingivalis, has made it a major pathogen of adult periodontitis [5].Immunohistological studies have established that a T cell/macrophage lesion identical to a delayed hypersensitivity 238 Effect of Fusobacterium nucleatum on the T and B cell responses toPorphyromonas gingivalis in a mouse model SUMMARYT cell cytokine profiles and specific serum antibody levels in five groups of BALB/c mice immunized with saline alone, viable Fusobacterium nucleatum ATCC 25586, viable Porphyromonas gingivalis ATCC 33277, F. nucleatum followed by P. gingivalis and P. gingivalis followed by F. nucleatum were determined. Splenic CD4 and CD8 cells were examined for intracytoplasmic interleukin (IL)-4, interferon (IFN)-gamma and IL-10 by dual colour flow cytometry and the levels of serum anti-F. nucleatum and anti-P. gingivalis antibodies determined by an ELISA. Both Th1 and Th2 responses were demonstrated by all groups, and while there were slightly lower percentages of cytokine positive T cells in mice injected with F. nucleatum alone compared with the other groups immunized with bacteria, F. nucleatum had no effect on the T cell production of cytokines induced by P. gingivalis in the two groups immunized with both organisms. However, the percentages of cytokine positive CD8 cells were generally significantly higher than those of the CD4 cells. Mice immunized with F. nucleatum alone had high levels of serum anti-F. nucleatum antibodies with very low levels of P. gingivalis antibodies, whereas mice injected with P. gingivalis alone produced anti-P. gingivalis antibodies predominantly. Although the levels of anti-F. nucleatum antibodies in mice injected with F. nucleatum followed by P. gingivalis were the same as in mice immunized with F. nucleatum alone, antibody levels to P. gingivalis were very low. In contrast, mice injected with P. gingivalis followed by F. nucleatum produced equal levels of both anti-P. gingivalis and anti-F. nucleatum antibodies, although at lower levels than the other three groups immunized with bacteria, respectively. Anti-Actinobacillus actinomycetemcomitans, Bacteroides forsythus and Prevotella intermedia serum antibody levels were also determined and found to be negligible. In conclusion, F. nucleatum ...

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Section: Immunization Proceduresmentioning

confidence: 99%

Section: Flow Cytometric Analysismentioning

confidence: 99%

Effect of Fusobacterium nucleatum on the T and B cell responses to Porphyromonas gingivalis in a mouse model

Gemmell

Bird

Carter

et al. 2002

Clinical and Experimental Immunology

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“…Serum was then separated and stored at À208C until used. Serum antibodies against heat-killed A. actinomycetemcomitans were assayed by ELISA as described previously [28]. Briefly, 96-well high-binding plates (Maxisorb Immunoplates, Nunc, Roskilde, Denmark) were coated with 50 ìl of antigen suspension (heat-killed A. actinomycetemcomitans 0:2 ìg=ml in 100 mM carbonate buffer).…”

Section: Elisa For Serum Antigen-specific Antibodiesmentioning

confidence: 99%

“…Pooled serum, derived from heat-killed A. actinomycetemcomitans-immunised mice, was used as positive control and PBS was used as negative control on each plate. Results were expressed as ELISA units, which were calculated by dividing the optical density of the sample by that of the positive control and multiplication by 100 [28]. …”

Section: Elisa For Serum Antigen-specific Antibodiesmentioning

confidence: 99%

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The role of macrophages in the induction of murine immune response to Actinobacillus actinomycetemcomitans

Sosroseno

Herminajeng

2002

Journal of Medical Microbiology

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Immunological Techniques: ELISA, Flow Cytometry, and Immunohistochemistry

Ford¹

2010

Methods in Molecular Biology

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Techniques to analyze the host immune response elicited by the presence of oral microorganisms and their products are central to our understanding of the local and systemic effects of oral diseases. This immune response has been extensively investigated for periodontal disease. The local response may result in lesions involving the gingival tissues and depending upon host susceptibility and microbial virulence may lead to local tissue destruction. More recently, however, the importance of the systemic inflammatory and immune response to oral organisms has been recognized. These systemic responses have been associated with an increased risk for cardiovascular disease, diabetes, and preterm low birth weight. A number of techniques are used extensively by researchers investigating humoral and cellular immune responses to oral organisms both in local oral tissues and fluids and systemically in peripheral blood. These are enzyme-linked immunosorbent assay (ELISA) to quantify specific antibody and cytokines in serum, gingival crevicular fluid (GCF), and saliva; characterization of T cells from peripheral blood and gingival tissues using flow cytometry; and immunohistological analysis of the inflammatory cell infiltrate in gingival tissues.

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Protective Immunity to Porphyromonas gingivalis Infection in a Murine Model

Cited by 47 publications

References 25 publications

Effect of Fusobacterium nucleatum on the T and B cell responses to Porphyromonas gingivalis in a mouse model

Effect of Fusobacterium nucleatum on the T and B cell responses to Porphyromonas gingivalis in a mouse model

The role of macrophages in the induction of murine immune response to Actinobacillus actinomycetemcomitans

Immunological Techniques: ELISA, Flow Cytometry, and Immunohistochemistry

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