David P. Cappelli

A novel Brucella isolate in association with two cases of stillbirth in non-human primates - first report.

Background  Brucellosis is veterinary and human health problem.

Characteristics and Utilization of Antibody Measurements in Clinical Studies of Periodontal Disease

The detection and quantitation of immune responses to infections have long been used as a diagnostic tool in medical infections. Recently, increasing evidence has supported that active, specific antibody responses to selected members of the subgingival microbiota are noted in periodontitis patients. This report describes the various specificities of this antibody as they relate to periodontitis classification and prognosis. The functional aspects of the serum antibody have come under increasing scrutiny to understand better the potential immunologic mechanisms acting in the periodontium. Data are available that describe opsonizing potential, complement fixing ability, blocking functions, and anti-toxic capacity for the antibody. Longitudinal alterations in specific antibody levels are shown to relate to infection and accompany changes in the burden of a specific microorganism in the subgingival plaque. Thus, these antibody changes could be useful indicators of altered host-parasite interactions that presage a disease-active episode. Finally, studies were designed to examine the ability of antibody to reflect the effects of treatment on the disease. The results indicated that specific antibody levels change with mechanical, antimicrobial, and anti-inflammatory treatments. The findings described in this report suggest that evaluation of the level and specificity of serum antibody can be a beneficial adjunct in designing and implementing clinical studies delineating the initiation, progression, and treatment of periodontitis. J Periodontol 1992; 63:1110-1116.

Periodontal diseases: to protect or not to protect is the question?

For decades, investigations have identified local and systemic humoral immune responses to microorganisms comprising the supra- and subgingival biofilms in the oral cavity. Inflammation and tissue destruction in the periodontium are accompanied by alterations in the quantity, quality, and specificity of antibody. The conundrum in this scenario is the existence of a substantial plasma cell infiltrate at sites of periodontal lesions and a seemingly robust antibody response in the oral cavity and the serum, apparently coincident with progressing disease. Consequently, much effort has been expended to elucidate the critical characteristics of protective humoral responses and to develop strategies for enhancing these unique features. We and others have conducted studies attempting to distinguish disease susceptibility associated with: i) variations in response levels-significantly increased to some species with disease, minimal response to others; ii) functional comparisons of antibody-subclass differences, genetic regulation, and maturation of responses; iii) microbial and antigenic specificity of the antibody-focus on specific pathogens and identification of selected antigens as targets for immunoprotection; and, iv) kinetics of responses during disease and therapeutic interventions-linking immune changes with infection and as a measure of treatment success. This report summarizes varied research designs and results, to provide a profile of antibody in health, gingivitis, and periodontitis. These profiles may be used to provide a framework focusing on the humoral response to commensal microorganisms and likely pathogens, as they emerge in the biofilm-etiologic for or in response to disease processes. Models for antibody as a diagnostic adjunct and for predicting protective antibody responses are suggested. These concepts are likely relevant for considering vaccine approaches to periodontitis.

Antigen Specificity of Serum Antibody in A. actinomycetemcomitans-infected Periodontitis Patients:

We hypothesized that serum antibody with selected antigen specificities would relate to infection and disease in the patients and, thus, describe the characteristics of potential protective antibody. This study used serum samples from 24 periodontitis patients with subgingival infection and elevated serum IgG antibody to A. actinomycetemcomitans to define the antigenic specificities of IgG, IgM, IgA, and IgGl-4 antibody to A. actinomycetemcomitans strain Y4 outer membrane antigens (OMA). Uniform IgG antibody (> 70% of the patients) was noted to antigens with Mr of 65, 38, 29, and 17 kDa. Both IgA and IgM specificities reflected those shown for IgG in each patient. IgGl and IgG2 antibody reacted with several OMA bands in each patient, while IgG3 antibodies were directed to numerous OMA bands in many patients and represented the most broad-based response. The IgG4 response patterns were limited to a few OMA bands. We noted a prominent occurrence of IgG reactions with OMA bands that were characteristic for individual patients. The frequency of responses to OMA of higher Mr (i.e., > 80 kDa) and to the 34-, 31-, and 24-kDa antigens was positively related to the total IgG antibody levels. Antibody reactive with OMA bands at 65-, 38-, 29-, 17-, 15-, and 11-kDa antigens was detected in patients with few to many teeth infected with A. actinomycetemcomitans. Furthermore, patients with a high percentage of teeth with ≥ 6 mm pockets had a decreased frequency of responses to the high-Mr antigens (i.e. > 90 kDa) as well as to the 58-kDa antigen. These findings indicate that human antibody reactivities with specific OMA bands of A. actinomycetemcomitans: (i) are positively correlated with the level of serum antibody, (ii) are associated with the number of teeth infected, and (iii) describe differences in the severity of the disease as measured by the frequency of teeth with deep pockets.

Systemic inflammatory responses in progressing periodontitis during pregnancy in a baboon model

This study tested the hypothesis that pregnant female baboons exhibit increased levels of various inflammatory mediators in serum resulting from ligature‐induced periodontitis, and that these profiles would relate to periodontal disease severity/extent in the animals. The animals were sampled at baseline (B), mid‐pregnancy (MP; two quadrants ligated) and at delivery (D; four quadrants ligated). All baboons developed increased plaque, gingival inflammation and bleeding, pocket depths and attachment loss following placement of the ligatures. By MP, both prostaglandin E2 (PGE2) and bactericidal permeability inducing factor (BPI) were greater than baseline, while increased levels of interleukin (IL)‐6 occurred in the experimental animals by the time of delivery. IL‐8, MCP‐1 and LBP all decreased from baseline through the ligation phase of the study. Stratification of the animals by baseline clinical presentation demonstrated that PGE2, LBP, IL‐8 and MCP‐1 levels were altered throughout the ligation interval, irrespective of baseline clinical values. IL‐6, IL‐8 and LBP were significantly lower in the subset of animals that demonstrated the least clinical response to ligation, indicative of progressing periodontal disease. PGE2, macrophage chemotactic protein (MCP)‐1, regulated upon activation, normal T cell expressed and secreted (RANTES) and LBP were decreased in the most diseased subset of animals at delivery. Systemic antibody responses to Fusobacterium nucleatum, Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Campylobacter rectus were associated most frequently with variations in inflammatory mediator levels. These results provide a profile of systemic inflammatory mediators during ligature‐induced periodontitis in pregnant baboons. The relationship of the oral clinical parameters to systemic inflammatory responses is consistent with a contribution to adverse pregnancy outcomes in a subset of the animals.

Role for Recombinant γ-Glutamyltransferase from Treponema denticola in Glutathione Metabolism

ABSTRACT Volatile sulfur compounds, including hydrogen sulfide (H2S), have been implicated in the development of periodontal disease. Glutathione is an important thiol source for H2S production in periodontal pockets. Our recent studies have delineated a pathway of glutathione metabolism in Treponema denticola that releases H2S. In this pathway, γ-glutamyltransferase (GGT) has been proposed to catalyze the first step of glutathione degradation. We have cloned the gene of GGT from T. denticola, which contains an open reading frame of 726 bp encoding a protein of 241 amino acids. Transformation of this gene into Escherichia coli led to the expression of a recombinant protein. After purification by chromatography, the recombinant protein showed enzymatic activity typical of GGT, catalyzing the degradation of Na-γ-glutamyl-4-nitroaniline (GNA) and the hydrolysis of glutathione, releasing glutamic acid or glutamine and cysteinylglycine. l-Cysteine is not a substrate of GGT. Importantly, GNA, when added to T. denticola, was able to compete with glutathione and inhibit the production of H2S, ammonia, and pyruvate. This was accompanied by the suppression of hemoxidative and hemolytic activities of the bacteria. Purified GGT was inactivated by TLCK (Nα-p-tosyl-l-lysine chloromethyl ketone) and proteinase K treatment. However, higher enzymatic activity was demonstrated in the presence of 2-mercaptoethanol and dithiothreitol. Our further experiments showed that the addition of recombinant GGT to Porphyromonas gingivalis, a bacterium without significant glutathione-metabolizing capacity, drastically increased the utilization of glutathione by the bacterium, producing H2S, ammonia, and pyruvate. This was again accompanied by enhanced bacterial hemoxidative and hemolytic activities. Together, the results suggest an important role for GGT in glutathione metabolism in oral bacteria.

Periodontitis in Pregnancy: Clinical and Serum Antibody Observations From a Baboon Model of Ligature-Induced Disease

BACKGROUND Chronic oral infections that elicit host responses leading to periodontal disease are linked with various sequelae of systemic diseases. This report provides seminal information on the clinical and adaptive immunologic characteristics of a baboon model of ligature-induced periodontitis during pregnancy. METHODS Female Papio anubis were evaluated for periodontal health at baseline. Ligatures were tied around selected teeth to initiate oral inflammation and periodontitis. Then the animals were bred. At midpregnancy ( approximately 90 days), a clinical evaluation was performed, and additional ligatures were tied on teeth in the contralateral quadrants to maintain progressing periodontitis throughout pregnancy. A final clinical evaluation was done for all experimental teeth after delivery, and ligatures were removed. Serum was collected at all sampling intervals for the determination of antibody levels to a group of 20 oral bacteria. Unligated animals served as controls. RESULTS At baseline, 16% of animals exhibited minimal plaque and gingival inflammation without periodontal disease. The remaining baboons demonstrated varying levels of inflammation/bleeding, and approximately 20% of the population had periodontal pocketing (>3 mm). Ligated animals expressed increased levels of inflammation and increased probing depths and clinical attachment loss (AL) and could be stratified into multiple subsets postligation based upon changes in clinical parameters at midpregnancy and at delivery. Baboons were categorized into disease susceptibility groups (periodontal disease susceptibility 1 through 4) that described the extent/severity of induced disease during pregnancy. Control animals showed minimal periodontal changes during gestation. Significant differences in serum antibody to multiple oral bacteria were found in animals presenting with periodontitis at baseline and during the 6 months of ligature-induced disease. A significant correlation to antibody to P. gingivalis, which was sustained throughout ligation and pregnancy, was observed with disease presentation. CONCLUSIONS The clinical presentation at baseline, reflecting the natural history of oral disease in these animals, suggests individual variation that is reflected in the characteristics of the adaptive immune responses to oral bacteria. The variability in the response to ligation with resulting periodontal disease provides a model to document prospectively the relationship between oral and systemic health outcomes.

Antigenic Specificity of Gingival Crevicular Fluid Antibody to Actinobacillus actinomycetemcomitans

Elevated antibody levels to periodontopathogens in GCF have been identified and used as support for local antibody synthesis in periodontitis. This study examined both cross-sectional and longitudinal GCF samples for the antigenic specificity of antibody in the fluid. GCF samples were collected from each tooth of 27 periodontitis patients infected with A. actinomycetemcomitans. Levels of IgG antibody in the GCF were assessed by means of an ELISA and compared with serum for determination of local elevations. A proportion of those GCF samples that exhibited significantly elevated antibody were examined by Western immunoblotting to outer membrane antigens from A. actinomycetemcomitans. Homologous sera were also examined for comparison of antibody specificities. Of the sites with elevated IgG antibody, 87% were colonized by A. actinomycetemcomitans; however, 46% of sites with A. actinomycetemcomitans infection did not have elevated antibody. Cross-sectional studies identified a 78 to 100% agreement between the antibody specificities in GCF and those in serum. Additionally, patterns of antibody reactivity in both GCF and serum in the subjects were often very distinctive. Longitudinal alterations in GCF antibody were examined in 15 patients through a monitoring interval of up to 2 years and showed a general conservation of specificities. However, 7/15 patients exhibited a definite acquisition of different antibody specificities during the monitoring. These results describe a relationship between elevated local antibody and A. actinomycetemcomitans infection. Furthermore, the antibody specificities in serum appear to reflect generally the local response to this pathogen.

Oral Consequences of Compromised Nutritional Well-Being

Dental structures are influenced by nutritional status only during the period of tooth formation. Thereafter, deficient nutrition may influence the supporting structures of the teeth and the oral mucosa Adequate early feeding programs in developing countries are important to avoid enamel defects and compromised salivary secretion, both of which may increase susceptibility to dental caries, the latter having more widespread oral consequences The early signs of nutritional deficiencies are seen in the oral soft tissues and include thinning, inflammation, and ulceration. Malnutrition also impairs immune responses and may predispose to life-threatening diseases of the oral soft tissues such as noma An optimal nutritional status is important in reducing the origin and severity of periodontal disease but is likely to be of limited value if the stimuli from dental plaque are not removed. Further research is needed in order to identify to what extent dietary modification will modulate periodontal disease and subsequent tooth loss Nutritional status and oral health are reciprocally related, and each one affects the other— a downturn in nutrition impairs oral function

Periodontitis in pregnant baboons: systemic inflammation and adaptive immune responses and pregnancy outcomes in a baboon model

BACKGROUND/OBJECTIVES Chronic periodontal infections have been suggested to contribute to the risk of adverse pregnancy outcomes. MATERIAL AND METHODS This study describes the relationship of patterns of systemic inflammatory mediators and IgG antibody to 20 oral bacteria in pregnant female baboons (Papio anubis) coupled with clinical features of ligature-induced periodontitis, as risk indicators for adverse pregnancy outcomes. Animals showing a preterm delivery and/or low birth weight newborns, as well as those pregnancies resulting in spontaneous abortion, stillbirth, or fetal demise were tabulated as adverse pregnancy outcomes. RESULTS A significantly greater frequency of the periodontitis group neonates had a low birth weight (18.1%; p = 0.008) and decreased gestational age (9.8%). Spontaneous abortion/stillbirth/fetal demise were increased in the periodontitis (8.7%) versus the control group (3.8%) (p = 0.054). The baseline oral clinical presentation of the experimental animals did not relate to the adverse pregnancy outcomes. Animals with the greatest extent/severity of periodontitis progression during the initial ½ of gestation (ie. to mid-pregnancy) had the greatest risk for adverse pregnancy outcomes. Baseline biological parameters indicating historical responses of the animals to periodontal challenge demonstrated individual variation in selected mediators, some of which became more differential during ligature-induced periodontitis. The relationship of clinical parameters to systemic inflammatory responses was consistent with a temporal contribution to adverse pregnancy outcomes in a subset of the animals. CONCLUSIONS These results support a link between periodontitis and adverse pregnancy outcomes in the baboons and provide a prospective experimental model for delineating the biologic parameters that contribute to a causal relationship between chronic oral infections and birth events.