Immunologic cross-reactivity between Streptococcus mutans and mammalian tissues.

The presence of cross-reacting antigens shared by oral streptococci and human and monkey tissue was investigated by indirect immunofluorescence. Rabbit antisera prepared against two strains of each of the seven serotypes of Streptococcus mutans were incubated on sections of cardiac muscle, skeletal muscle, kidney, brain, liver, and skin. Antisera to S. mutans, serotype g, strain OMZ-65 reacted with cardiac muscle components. This reactivity could be reduced by absorption with bacterial cell membranes. Antisera to S. mutans serotype g, strain K-1 reacted with both cardiac and skeletal muscle but could not be reduced by absorption with bacterial cells, cell walls, or cell membranes. Antisera to S. mutans serotype g, strain OMZ-65 reacted with kidney glomeruli and could be completely absorbed by a cell wall preparation. S. mutans, serotype e, strain MT703 also reacted with kidney glomeruli but could not be reduced by absorption with bacterial antigens.

Immunofluorescence of biopsy specimens: comparison of methods of transportation.

Immunofluorescence findings for IgG, IgA, IgM, and C4 on biopsy specimens quick frozen and transported frozen were compared with findings on portions of the same biopsy specimens placed in holding solution and transported at ambient temperatures. A total of 52 biopsy specimens were examined from normal individuals and patients with systemic lupus erythematosus (SLE), discoid lupus erythematosus (DLE), pemphigus, pemphigoid, and dermatitis herpetiformis. Overall agreement of results was 90%: 88% in SLE; 50% to 66% in DLE, 96% in pemphigus; 92% in bullous pemphigoid; and 87% in dermatitis herpetiformis. Except for two of the 42 biopsy specimens, the combined IgG, IgA, IgM, and C4 findings were the same. In one DLE case, only the frozen biopsy specimen was positive. In one case of dermatitis herpetiformis, only the ambient temperature biopsy specimen was positive. Results indicate the holding solution at ambient temperature can be used in place of the frozen method.

Diagnosis of systemic lupus erythematosus. Importance of antinuclear antibody titers and peripheral staining patterns.

Titers and patterns of antinuclear antibodies (ANA) in sera from 134 normal blood donors, 20 patients with rheumatoid arthritis, 15 patients with systemic scleroderma, and 32 patients with diagnosed or suspected systemic lupus erythematosus (SLE) were studied. The difference between the findings with sera of patients with SLE and normal subjects in terms of high (greater than 160) titers of ANA was greater than in terms of peripheral staining patterns. However, in comparing sera from patients with SLE with sera from patients with other connective tissue diseases, greater differences were found in the incidence of peripheral patterns of ANA compared to differences in the frequency of high ANA titers. Maximum specificity in the diagnosis of SLE was achieved when both titers and patterns of ANA were considered.

Sjogren's syndrome and keratoconjunctivitis sicca.

Keratoconjunctivitis sicca patients diagnosed on the basis of a history, dry-eye symptoms, and definite clinical signs of keratoconjunctivitis sicca, with the associated symptoms of dry mouth and/or arthritis, had measurements of tear osmolarity, Schirmer tear test without anesthetic, stimulated parotid salivary flow, and serum analysis for the presence of autoantibodies associated with Sjogren's Syndrome. In contrast to previous studies, a lower incidence of SS-A or SS-B (1-3%), ANA (41-47%), DNA (11-16%), and RF (9-12%) serum antibodies was detected. Salivary-stimulated parotid flow was abnormally decreased in 59% of the patients. Sjogren's syndrome, as indicated by the presence of serum antibodies, appears to have a lower incidence in keratoconjunctivitis sicca than considered previously.

The treatment of desquamative gingival lesions.

The treatment of desquamative gingival lesions is dependent on the correct diagnosis of the underlying disease, which is frequently dermatologic. This is determined by clinical observations, as well as by histologic and immunologic examination of gingival biopsy specimens. The recommended treatment of desquamative lesions caused by lichen planus, cicatricial pemphigoid (benign mucous membrane pemphigoid), pemphigus, and psoriasis, the most common dermatoses causing gingival lesions, is summarized on the basis of 62 cases and reports in the literature.

Desquamative lesions of the gingiva.

Previous reports, as well as the investigations reported here, clearly indicate that desquamative gingivitis is a clinical manifestation of several diseases. Correct identification of the underlying etiology is very important since approximately one-third of the patients with desquamative gingivitis may have cicatricial pemphigoid or pemphigus. These two diseases have broad and sometimes systemic medical implications. In cicatricial pemphigoid, not only the oral mucosa but also the conjunctiva may be involved with subsequent blindness. Because of this, patients with cicatricial pemphigoid should also be examined by an ophthalmologist and may require care of a dermatologist for systemic treatment. In pemphigus, 50% of the cases start with only oral lesions with later development of skin lesions. Because of the life-threatening nature of this disease, patients are usually placed on high systemic doses of corticosteroids. Patients with pemphigus should be referred to a dermatologist immediately.

Desquamative gingivitis: immunologic findings.

A case of desquamative gingivitis is presented that was a gingival manifestation of cicatricial pemphigoid. Immunologic studies of serum and biopsies were of diagnostic significance. The application of immunofluorescence for the diagnosis of desquamative gingival lesions was discussed. If a definite diagnosis can not be made on the basis of light microscopy, immunofluorescence studies should be performed.

Bacterial IgG and IgM antibody titers in acute necrotizing ulcerative gingivitis.

IgG AND IgM ANTIBODY TITERS to eight bacterial isolates were measured by indirect immunofluorescence and ELISA in sera from acute necrotizing ulcerative gingivitis (ANUG) patients during the acute phase, from ANUG patients during the convalescent phase, from patients with gingivitis and from subjects with normal gingiva. Subjects were matched with respect to age and sex. Compared to the gingivitis and healthy groups, the ANUG groups exhibited significantly higher IgG and IgM titers to intermediate-sized spirochetes and higher IgG titers to Bacteroides melaninogenicus subsp intermedius. The findings support recent studies showing that these organisms are major bacterial components in ANUG lesions. They also suggest that these bacteria proliferate above-normal levels several weeks or months prior to the clinical onset of ANUG.

The alteration in gingival basement membrane antigens in chronic periodontitis.

The gingival basement membrane antigens, Type IV collagen, bullous pemphigoid antigen and epidermolysis bullosa acquisita antigen were studied by indirect immunofluorescence in 11 gingival specimens from patients with periodontitis and 2 normal gingival specimens. In the normal control gingival specimens, the antigens were all present and stained with a continuous linear pattern. In periodontitis, alterations occurred in the gingival basement membrane antigens in the apical portion of the pockets. These included thinning, interruptions, partial or complete absence involving one or more rete pegs and fragmentation. These alterations may result form the disease process or play a role in the pathogenesis.

The regeneration of gingival basement membrane antigens during secondary wound healing.

The reformation of basement membrane antigens was examined during healing following gingivectomy procedures. Three antigens, Type IV collagen, bullous pemphigoid antigen and epidermolysis bullosa acquisita (EBA) antigen, were identified by indirect immunofluorescent tests. While all three antigens could be identified in the healing wound within 2 days, the antigens regenerated at different rates. The bullous pemphigoid antigen reformed the earliest and extended furthest along the healing front of epithelium. The Type IV collagen reformed at an intermediate rate and the EBA antigen formed the slowest. These results confirm that these three antigens are different and suggest that the gingival basement membrane is a heterogeneous structure which contains elements that form at different rates during secondary wound healing.

Experimental periodontal disease. Immediate hypersensitivity.

Immediate hypersensitivity reactions were induced in the periodontium and skin of monkeys sensitized with novo alcalase. Animals were challenged in the gingival papillae with 0.1, 1.0, and 10 micrograms of antigen for 1, 3, 5, or 7 consecutive days prior to sacrifice. At the same time, skin sites were challenged with 1 microgram of antigen. With repetitive immediate hypersensitivity reactions, the inflammatory infiltrate changed from one characterized by polymorphonuclear leukocytes to one characterized by plasma cells and lymphocytes. The repetitive gingival exposure to bacterial antigens which occur in periodontal disease could lead to repetitive immediate hypersensitivity reactions. Such reactions could play a role in the histopathology of human periodontal disease.

Development of a rapid latex agglutination test for periodontal pathogens.

The studies reported here describe the development, characterization, and initial application of latex agglutination assays for periodontal pathogens. Latex reagents were prepared by sensitization of latex microspheres with rabbit IgG antibodies to either Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, or Prevotella intermedia. The protein concentration utilized for sensitization and microsphere size were optimized. One reagent was prepared to A. actinomycetemcomitans and a second combination reagent was prepared by mixing latex sensitized with antibodies to P. gingivalis and latex sensitized with antibodies to P. intermedia. The sensitivity of both latex reagents in the traditional wet and a dried format was evaluated. In addition, sensitivity and specificity with homologous and heterologous bacterial suspensions were evaluated. The reagents were found to demonstrate both specificity and sensitivity. Initial studies with subgingival human plaque demonstrated the ability of these reagents to detect the specific organisms in plaque.

Humoral immunologic responses in idiopathic juvenile periodontitis (periodontosis).

Humoral responses were examined in idiopathic juvenile periodontitis (IJP) including antibody titers to representative strains of the five groups of periodontosis-associated bacteria. Titers as determined by indirect immunofluorescence were compared in IJP, non-IJP family members, periodontitis and periodontally healthy subjects. Serum concentrations of IgG, IgA, IgM and IgE were also assayed. Antibody titers to the periodontosis-associated bacteria were generally low. Some disease specificity was observed in that titers to Groups II and III bacteria were highest in IJP. These titers in IJP were significantly higher (P less than 0.01) than in the other subjects. Titers to Group I bacteria were similar in all periodontal groups except the periodontally healthy subjects where titers were the lowest. Titers to Group I bacteria were significantly higher (P less than 0.01) than in the periodontally healthy subjects. Titers to Groups IV and V were uniformly low in all periodontal groups. Immunoglobulin concentrations of IgA, IgM and IGE were similar in IJP and non-IJP. The IgG concentrations in IJP, while within the normal range were significantly higher in IJP compared to non-IJP family members.

Bacterial antibody titers in ligature-induced periodontitis in beagle dogs.

Serum antibodies to indigenous bacteria in ten beagle dogs were examined over a 7-month period during the development of ligature-induced periodontitis. Gram-negative strains comprised approximately 75% of the cultivable periodontitis microflora with a predominance of black-pigmented Bacteroides species. A total of 44 bacterial strains representing the predominant cultivable subgingival beagle dog microflora was selected for antibody determination. The IgG and, in some cases, IgM serum antibody titers to these organisms were determined by indirect immunofluorescence. The antibody titers to most test strains remained unchanged during the experimental period. Gram-negative bacteria generally exhibited lower titers than the Gram-positive bacteria. Especially low titers were found for the black-pigmented Bacteroides. Four dogs that developed the most severe periodontitis showed about 2-fold higher IgG titers to some Gram-negative anaerobic rods in the pre-ligation period than dogs that developed a more moderate periodontitis. These data suggested a possible diagnostic value of such antibody determinations. However, the overall finding of the present study was that serum antibody titers to key periodontopathic organisms remained low throughout the experiment. This result may suggest that the rapid periodontal destruction in ligature-induced periodontitis is due in part to an inadequate antibody response against the infecting microorganisms and their pathogenic products.

Actinobacillus actinomycetemcomitans serotypes and leukotoxicity in Korean localized juvenile periodontitis.

Actinobacillus actinomycetemcomitans is though to play an important role in the pathogenesis of localized juvenile periodontitis (LJP). Preliminary data suggested that the serotype distribution of A. actinomycetemcomitans in Korea and the United States differ. This study evaluated A. actinomycetemcomitans prevalence, serotype distribution, and leukotoxicity in Korean LJP patients by culture, enzyme-linked immunosorbent assay, indirect immunofluorescence, and lactate dehydrogenase release from polymorphonuclear leukocytes exposed to A. actinomycetemcomitans. A. actinomycetemcomitans occurred in 75% of LJP lesions and 6% of normal sites with approximately equal distribution of serotype a, b, and c. Single serotypes were isolated from nine patients while three patients harbored two serotypes either in the same or different disease sites. A. actinomycetemcomitans leukotoxicity occurred in 22% isolates with a 69% prevalence. Individual sites harbored both leukotoxic and non-leukotoxic strains with no serotype association. The distribution of serotypes and leukotoxic strains of A. actinomycetemcomitans in Korean LJP patients differed from those reported in the United States. This suggests that serotype b may not be more important in the pathogenesis of LJP.

Antibodies to Actinobacillus actinomycetemcomitans in a Korean population.

Previous studies demonstrated that serum antibodies to Actinobacillus actinomycetemcomitans strain Y4 are significantly elevated in sera from localized juvenile periodontitis (LJP) and postlocalized juvenile periodontitis (P-LJP) patients compared to normal controls in United States populations. This study examined the age of subjects in relation to the antibody levels to A. actinomycetemcomitans in a Korean population. Seven groups were investigated including sera from newborns, infants, children, periodontally normal puberty and adult groups and LJP and P-LJP groups. Antibody levels were determined by an enzyme-linked immunosorbent assay (ELISA) to A. actinomycetemcomitans strain SNUDC 10-1 (serotype C) isolated from a Korean LJP patient. In the healthy non-LJP and non-P-LJP subjects, IgG levels decreased from the newborn group to the 5-month-old group and then gradually increased through the adult group. The IgM levels in these groups continuously increased from birth until a plateau was reached in the 2- to 6-year group. Serum IgA levels to strain SNUDC 10-1 were too low to be measured by this assay.

Clinical and microbiological status of osseointegrated implants.

Peri-implantitis, an inflammatory response around implants, has a poorly defined etiology and pathogenesis. To better understand the role of specific microorganisms in this disease process, clinical and microbiological parameters were examined in 24 patients with 98 osseointegrated implants. Sites were evaluated for probing depth (PD), plaque/calculus index (PI), gingival bleeding index (GBI), mobility, and crevicular fluid flow rate (CFFR). Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia in subgingival plaque were identified by latex agglutination assays. Clinically, a statistically significant correlation (P < 0.001) was observed between probing depth and the length of time an implant was present. Mobility was also significantly greater (P < 0.001) in the maxillary than in the mandibular implants. Subgingival sites harboring one of the three microorganisms had significantly greater PD, GBI, and CFFR than non-colonized sites. Implants in partially edentulous patients more frequently were colonized with P. gingivalis/P. intermedia than edentulous patients. The incidence of these microorganisms also correlated with fixture longevity. Implants present for 3 to 4 years had a significantly greater frequency of test microorganisms than implants present for 1 to 2 years. These findings suggest that microbial pathogens associated with periodontitis occur more commonly around implants exhibiting gingival inflammation (GBI) and may contribute to peri-implantitis.

Humoral antibody responses in periodontal disease.

Several forms of periodontal disease are considered to be infectious diseases with associated specific bacteria. This study examined the humoral antibody levels as assayed by ELISA to Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia in adult periodontitis (AP), localized juvenile periodontitis (LJP), rapidly progressive periodontitis (RPP), and in periodontally healthy subjects (HS). Sixty-two of the 64 (96.9%) patients had significantly elevated antibody levels to at least one of the three organisms. Elevated levels of antibodies to P. gingivalis occurred in 82.8% of the RPP, LJP, and AP patients with all 3 disease groups showing greater responses than HS controls. Antibodies to A. actinomycetemcomitans were found in 59.4% of the RPP, LJP, and AP patients and were significantly higher in both LJP and RPP patients. Only 21.9% of the RPP, LJP, and AP patients showed elevated levels to P. intermedia with only significantly higher levels in the RPP and LJP groups. Antibodies to A. actinomycetemcomitans and P. intermedia were rarely found alone (only 5.1% and 2.6% of the patients respectively) but were usually accompanied by P. gingivalis. These results suggest that one or more combinations of these 3 bacteria may play a role in the pathogenesis of these forms of periodontal disease.

The presence of bacteria in the oral epithelium in periodontal disease. III. Correlation with Langerhans cells.

Langerhans cells (LC) are cell types found in the skin and gingiva. LC have immunological functions as phagocytic cells and as antigen-presenting cells for T and B lymphocytes. Sections from biopsies of the gingiva in cases of periodontal disease were found to have increased numbers of LC. These biopsies also contained intragingival bacteria. Serial sections of frozen specimens of human gingiva were prepared for staining. Hematoxylin and eosin were used for tissue survey, the Gram stain for assessment of bacterial invasion, anti-Leu-6 monoclonal antibody associated with peroxidase technique (PAP) to identify LC, antibacterial sera to Bacteroides gingivalis and Actinobacillus actinomycetemcomitans associated with peroxidase to specifically identify these two common periodontopathogenic bacteria. Additional positive identification of bacteria was performed by preparing the same histological section containing gram-stained particles for scanning electron microscope and transmission electron microscope LC confirmation. The results suggest that the increased number of LC seen in diseased sites of oral epithelium containing intragingival microorganisms may be one of the host immune mechanisms to penetration by bacteria.

Clinical and microbiological effects of minocycline-loaded microcapsules in adult periodontitis.

Clinical and microbiological effects of subgingival delivery of 10% minocycline-loaded (MC), bioabsorbable microcapsules were examined in 15 adult periodontitis patients. Patients received oral hygiene instruction 2 weeks prior to the study. At baseline (day 0) all teeth received supragingival scaling (SC); 2 quadrants received no further treatment and 1 quadrant received subgingival scaling and root planning (SRP). In the fourth quadrant, the tooth with the deepest probing sites (at least 1 site > or = 5 mm) was treated with minocycline microcapsules. The sites were evaluated at baseline and weeks 1, 2, 4, and 6. Clinical indices included bleeding on probing (BOP), probing depths (PD), and attachment loss (AL). Microbiological evaluations included percent morphotypes by phase-contrast microscopy; cultivable anaerobic, aerobic, and black-pigmented Bacteroides (BPB); and percent Porphyromonas gingivalis, Prevotella intermedia, Eikenella corrodens, and Actinomyces viscosus by indirect immunofluorescence. In the SC + MC group, BOP, PD, and AL were significantly reduced from baseline for weeks 1 to 6. BOP in the SC + MC group was significantly reduced compared to the SRP group from weeks 2 to 6. In the SC + MC group the percent of spirochetes and motile rods decreased and the percent of cocci increased after 1 week. The increased cocci and decreased motile rods were statistically greater at weeks 4 and 6 in the SC + MC group compared to the SRP group. This study demonstrates that local subgingival delivery of 10% minocycline-loaded microcapsules as an adjunct to scaling results in reduction in the percent sites bleeding on probing greater than scaling and root planning alone and induces a microbial response more favorable for periodontal health than scaling and root planing.