Interaction Between Immune Cells and Bacteria Associated With Periodontitis

Study Overview
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Brief Summary
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This study evaluates the interaction between host immune cells and bacteria associated with periodontitis. It comprises biological material from donors with and without periodontal disease. Specifically, we collect a spit and blood sample to conduct in vitro stimulations and measurements of selected parameters related to periodontitis to clarify obscure areas in the immunologic pathogenesis of this disease.

Detailed Description
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Periodontitis is a prevalent, multifactorial inflammatory disease characterized by the interaction between microorganisms organized in biofilms on tooth surfaces and host immune cells, leading to an inflammatory destruction of the tooth-supporting tissues and - if left untreated - eventually tooth loss. Periodontitis affects up to 50% of the population in the United States of America, and is classified in an aggressive and a chronic form depending on genetic factors, age of onset, speed and severity of attachment loss. The onset of periodontitis is caused by an immunologic imbalance between host immune cells and residing microorganisms in subgingival pockets. The host immune cells are capable of enhancing both a protective and a destructive inflammatory response towards the microorganisms through the release of inflammatory mediators e.i. proinflammatory and antiinflammatory cytokines. The role of antibodies in periodontitis is also unclear. Some studies show an excessive antibody level against bacteria associated with periodontitis e.g. Porphyromonas gingivalis (P.g.). In general, this study contributes to a profound understanding of the host immune cells role in the onset and pathogenesis of periodontitis by comparing healthy versus diseased donors immunologic responses toward pathogene and apathogene microorganisms and their genetic background.

Official Title
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Interaction Between Immune Cells and Bacteria Associated With Periodontitis

Conditions
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Periodontal Diseases, Periodontitis, Aggressive Periodontitis, Immunologic Disease, Microbial Disease, Periodontal Pocket, Inflammation, Inflammation Gum, Dysbiosis, Rheumatoid Arthritis, Generalized Aggressive Periodontitis, Generalized Chronic Periodontitis, Chronic Periodontitis

Intervention / Treatment
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* Other: In vitro stimulation of blood with periodontitis-associated- and control bacteria
* Diagnostic Test: Anti-CCP- and anti-P.g.-antibodies titers
* Genetic: Analysis of selected single nucleotide polymorphisms (SNPs)
* Diagnostic Test: periodontitis-associated bacteria presence


Participation Criteria
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Eligibility Criteria
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Inclusion criteria for chronic periodontitis donors: 50-60 years of age. Interproximal attachment loss at minimum 3 teeth besides molars and incisors. Clinical attachment loss at minimum 10 sites identified by bleeding and pus upon probing. Visible radiographic bone loss. Medically healthy donors. Inclusion criteria for aggressive periodontitis donors: 19-40 years of age. Interproximal attachment loss at minimum 3 teeth besides molars and incisors. Clinical attachment loss at minimum 10 sites identified by bleeding and pus upon probing. Visible radiographic bone loss. Medically healthy donors. Inclusion criteria for healthy donors: (age: 19-40 years; 50-60 years) No sign of inflammatory conditions or other general systemic diseases. Medically healthy donors. Exclusion criteria for all groups: Pregnant and breastfeeding. Antibiotic treatment within 6 months. Suffer from periodontal manifestations caused by systemic diseases e.i. genetic diseases, haematologic anomalies or syndromes.

Ages Eligible for Study
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Minimum Age: 19 Years
Maximum Age: 60 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Chronic periodontitis donors<br>Donors are medically healthy. Slow to moderate attachment loss and bone destruction. Good correlation between etiological factors and serverity of attachment loss. | Other: In vitro stimulation of blood with periodontitis-associated- and control bacteria<br>* Peripheral mononuclear blood cells are stimulated with periodontitis-associated- and control bacteria to measure the amount of positive cytokine-producing cells.<br>Diagnostic Test: Anti-CCP- and anti-P.g.-antibodies titers<br>* Anitbody titers will be measured in saliva and serum samples.<br>Genetic: Analysis of selected single nucleotide polymorphisms (SNPs)<br>* DNA obtained from saliva samples will be used to determine the genotype of the participants for selected SNPs.<br>Diagnostic Test: periodontitis-associated bacteria presence<br>* Determination of the presence of periodontitis-associated bacteria e.i. Porphyromonas gingivalis in saliva and blood samples.<br>|
| Aggressive periodontitis donors<br>Donors are medically healthy. Rapid attachment loss and bone destruction. Familial aggregation. No correlation between etiological factors and serverity of attachment loss. | Other: In vitro stimulation of blood with periodontitis-associated- and control bacteria<br>* Peripheral mononuclear blood cells are stimulated with periodontitis-associated- and control bacteria to measure the amount of positive cytokine-producing cells.<br>Diagnostic Test: Anti-CCP- and anti-P.g.-antibodies titers<br>* Anitbody titers will be measured in saliva and serum samples.<br>Genetic: Analysis of selected single nucleotide polymorphisms (SNPs)<br>* DNA obtained from saliva samples will be used to determine the genotype of the participants for selected SNPs.<br>Diagnostic Test: periodontitis-associated bacteria presence<br>* Determination of the presence of periodontitis-associated bacteria e.i. Porphyromonas gingivalis in saliva and blood samples.<br>|
| Control donors<br>Donors are medically healthy. No sign of inflammatory conditions. | Other: In vitro stimulation of blood with periodontitis-associated- and control bacteria<br>* Peripheral mononuclear blood cells are stimulated with periodontitis-associated- and control bacteria to measure the amount of positive cytokine-producing cells.<br>Diagnostic Test: Anti-CCP- and anti-P.g.-antibodies titers<br>* Anitbody titers will be measured in saliva and serum samples.<br>Genetic: Analysis of selected single nucleotide polymorphisms (SNPs)<br>* DNA obtained from saliva samples will be used to determine the genotype of the participants for selected SNPs.<br>Diagnostic Test: periodontitis-associated bacteria presence<br>* Determination of the presence of periodontitis-associated bacteria e.i. Porphyromonas gingivalis in saliva and blood samples.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| periodontitis-associated- and control bacterial stimulation of host immune cells. | Identification and determination of the amount of cytokine-producing immune cells when stimulated with bacteria associated with periodontitis including pro- and antiinflammatory cytokines. Genuses: Porphyromonas, Prevotella, Eikenella, Aggregatibacter, Actinomyces, Lactobacillus, Bifidobacterium, Rothia. | Aug. 2020 | 
| Anti-cyclic citrullinated peptides (anti-CCP) antibodies titers. | Determination of the prevalence of anti-CCP-positive periodontitis patients through measure of anti-CCP antibody titers in serum samples and correlation with the level of antibodies to P. gingivalis, and to the abundancy of the bacterium in saliva. | Aug. 2020 | 
| P. gingivalis presence and related antibodies. | Determination of P. gingivalis presence in saliva and serum samples through RT-qPCR and determination of the level of antibodies towards the bacterium using in-house Luminex-based technology. | Aug. 2020 | 
| Single nucleotide polymorphism (SNP) analysis. | Investigation of the potential association between periodontitis and selected polymorphisms in the PADI genes using multiplex bead-based SNP assays with the Luminex technology. | Aug. 2020 | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Cytokine profile in saliva. | Determination of the cytokine profile in saliva samples from subject with periodontitis and healthy controls with Luminex based technology. | Aug. 2020 | 
| Presence of other periodontal bacteria. | Determination of the presence of other periodontal bacteria through RT-qPCR assays. | Aug. 2020 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Periodontal disease, Periodontitis, Aggressive periodontitis, Dysbiosis, Inflammation, Regulation, Cytokines, Bacteria, Porphyromonas gingivalis, Saliva, Anti-CCP, Peptidyl arginine deiminases, Single nucleotide polymorphisms