Antibodies to mycobacterial stress protein in patients with orofacial granulomatosis.

Immunoglobulin G antibody titers to the mycobacterial stress protein with molecular weight of 65 kDa (mSP65) were determined by ELISA in sera from 10 patients with orofacial granulomatosis (OFG). Four patients with confirmed Crohn's disease had serum antibodies to mSP65 with titers ranging from 400-950. Of six remaining patients, three patients had serum antibodies to mSP65, with titers ranging from 180-850, whilst no serum antibody to this antigen could be detected in 3 patients. However, other laboratory investigations failed to show any consistent pattern of disturbance in the 10 tested. Thus, the presence of serum antibody to mycobacterial 65 kDa stress protein might prove to be of a diagnostic value for Crohn's disease.

Elevated IgA salivary antibody levels to mycobacterial 65 kDa stress protein in patients with oral candidosis.

Salivary IgA antibody levels to cytoplasmic protein extract of Candida albicans (CSE) and to mycobacterial 65 kDa stress protein (mSP65) were determined by ELISA in 25 patients with chronic atrophic oral candidosis and in 23 controls with clinically healthy oral mucosa. The results showed significantly elevated anti-CSE and anti-mSP65 antibody levels in patients when compared with healthy controls (P < 0.001). A statistically significant relationship was found between salivary antibody titres to CSE but not to mSP65 and C. albicans colony counts in patients with oral candidosis.

The immunogenicity of outer membrane proteins of haemin-depleted Porphyromonas (Bacteroides) gingivalis W50 in periodontal disease.

The antigens from outer membrane protein extracts of Porphyromonas gingivalis (W50), grown under different haemin concentrations, were examined for binding with serum antibodies from patients with severe progressive periodontitis or from periodontally healthy control subjects. P. gingivalis was grown under haemin limitation (0.33 micrograms/ml) and haemin excess (2.5 micrograms/ml) conditions in a chemostat at a mean generation time of 6.9 h, at pH 7.5. Sarkosyl-insoluble fractions of outer membrane proteins from P. gingivalis were prepared, and analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot techniques. The SDS-PAGE analysis of the outer membrane of haemin-limited P. gingivalis identified several new protein components, or changed expression of bands compared with cells grown under haemin excess. Immunoblot analysis showed IgG antibodies to 2 haemin deprivation-induced proteins in patients with severe progressive periodontitis, but not in the control sera. These results confirm the immunogenicity of some of the haemin-regulated outer membrane proteins of P. gingivalis in severe progressive periodontitis.

Serum zinc levels in patients with burning mouth syndrome.

The serum zinc levels were measured by atomic absorption spectrophotometry in 30 patients with burning mouth syndrome, (BMS) and in 30 control subjects with clinically healthy oral mucosa. The mean value of serum zinc levels in the patient group was found to be significantly lower (mean +/- SD = 12.13 +/- 1.40 mumol/L) than the mean value of serum zinc levels in the control group (mean +/- SD = 12.89 +/- 1.62 mumol/L; p less than 0.05). The number of subjects with serum zinc levels less than the minimum normal value (less than 11 mumol/L) was higher in the patients with BMS (30%) than in the control subjects (10%). These results suggest that in some patients low serum zinc levels may be associated with BMS. However, the number of patients with BMS was too small to allow definite conclusions.

T cell proliferative responses to molecular fractions of periodontopathic bacteria.

Soluble antigenic preparations of Veillonella parvula and Bacteroides gingivalis were separated by SDS-PAGE and used after electroblotting and solubilization for in vitro lymphocyte stimulation in 13 patients with severe periodontitis and 12 controls. The cellular responses of controls and patients to V. parvula antigens were represented by four main proliferation-inducing fractions with 74-66, 52-46, 22-19 and 12 kD mol. wt. These fractions induced slightly enhanced DNA synthesis in lymphocytes from eight patients who failed to respond to whole antigenic extract. Lymphocyte samples from Veillonella whole extract unresponsive patients were also examined for in vitro proliferation by B. gingivalis fractions. Almost all stimulatory activities could be classified into five regions of 84-74, 35-31, 28-25, 17-15 and 12 kD.

Elevated antibody levels to mycobacterial 65-kDa stress protein in patients with superficial candidiasis.

Antibody levels to the mycobacterial 65-kDa stress protein (mSP65) were determined by ELISA in sera from patients with chronic atrophic oral candidiasis, vulvovaginal candidiasis, Candida albicans-infected or noninfected oral lichen planus, or recurrent aphthous ulceration and from subjects with clinically healthy oral mucosa. The results showed significantly elevated anti-mSP65 antibody levels in patients with oral or vulvovaginal candidiasis and in patients with Candida-infected lesions of lichen planus when compared with patients with noninfected lichen planus or recurrent oral ulceration and with matched healthy controls (P less than .001). Immunoblot analysis showed that the rabbit antiserum that strongly bound to mSP65 cross-reacted only weakly with a homologous band of a soluble C. albicans extract. Moreover, the binding of antibodies from patients with candidiasis to the mSP65 antigen was not inhibited in the presence of Candida extract. In view of the poor serologic cross-reactivity, it seems plausible that the recall stimulation of anti-mSP65-producing B cells could be induced by helper T cells that cross-react with the structurally homologous protein of C. albicans.

Identification of immunodominant antigens of Candida albicans in patients with superficial candidosis.

The aims of this study were to identify the immunodominant protein antigens of Candida albicans in patients with superficial infections of the oral and vaginal mucosa. Cytoplasmic protein extract from C. albicans was analyzed by the immunoblot technique using sera from 20 patients with chronic atrophic oral candidosis, from 8 patients with vulvovaginal candidosis, and from 20 control subjects. A significantly greater proportion of patient sera reacted with the 65- and 32-kDa antigens when compared with sera from controls (P less than 0.001). Sera from patients also reacted more often with 38- and 29-kDa antigens (P less than 0.05), while sera from both patients and controls reacted with the 47-kDa antigen. The identified 65-, 38-, 32-, and 29-kDa antigens may be of importance for the development and also for the recurrence of superficial candidosis of the oral and/or vaginal mucosa.

Detection of immunodominant antigens of periodontopathic bacteria in human periodontal disease.

Sonicated whole cell extracts and outer membrane proteins (OMP) from Bacteroides gingivalis and Veillonella parvula were analysed by the immunoblot technique using sera from 103 patients with various forms of periodontal disease and from 31 control subjects. B. gingivalis sonicate contained 12 major bands (75-14 kDa) of which the 46, 27 and 14 kDa antigens reacted more frequently with sera from adult and young adult patients with severe periodontitis compared with sera from controls and mild periodontitis patients. The OMP of B. gingivalis contained 6 main antigens of 75, 57, 51, 46, 35 and 19 kDa m.w. The 46 kDa antigen reacted predominantly with sera from both groups of patients with severe periodontitis. V. parvula sonicate contained 11 antigens (76-13 kDa) of which the 76 kDa antigen reacted more frequently with sera from controls and patients with mild periodontitis than with sera from patients with severe periodontitis. Conversely, antibodies to the 39 kDa antigen (absent from OMP) were specifically associated with severe periodontitis. Further monitoring of antibody responses to the 46, 27 and 14 kDa antigens of B. gingivalis and 39 kDa antigen of V. parvula may be of importance for the assessment of severity of human periodontal disease.

Antigens of Actinobacillus actinomycetemcomitans identified by immunoblotting with sera from patients with localized human juvenile periodontitis and generalized severe periodontitis.

Sonicated whole cell extracts and outer membrane proteins from this bacterium were analysed using sera from 31 young patients with localized juvenile periodontitis, 55 young adults with generalized severe periodontitis and from 31 healthy control subjects. The sonicate contained 13 major bands (14-78 kDa); a greater proportion of sera from patients with generalized periodontitis reacted with 40 and 70 kDa antigens when compared with sera from localized juvenile periodontitis and controls. In contrast, a lower proportion of sera from localized juvenile periodontitis reacted with the 29 kDa antigen when compared with severe periodontitis and controls. The outer membrane proteins contained four main antigens of 19, 24, 35 and 67 kDa, which reacted with sera from all three groups. Although, so far, the findings do not allow discrimination between the two diseases, antibody responses to the 29, 40 and 70 kDa antigens of A. actinomycetemcomitans may help in the assessment of severity of the disease in patients with periodontitis.

In vitro lymphocyte proliferation by carbamazepine, carbamazepine-10, 11-epoxide, and oxcarbazepine in the diagnosis of drug-induced hypersensitivity.

In vitro lymphocyte proliferation induced by carbamazepine (CBZ) was evaluated in nine patients with hypersensitivity to this drug. Lymphocytes from all hypersensitive patients responded by significantly enhanced DNA synthesis to CBZ when patients were compared with 33 tested control subjects. However, lymphocytes from five of six hypersensitive patients were not stimulated by carbamazepine-10, 11-epoxide, and oxcarbazepine (OXC), and this was confirmed clinically in two CBZ hypersensitive patients with OXC therapy. The results indicate that OXC may be suitable as an alternative therapy in some patients with CBZ hypersensitivity.

Immunoglobulin A1 and A2 subclass of salivary antibodies to Candida albicans in patients with oral candidosis.

Immunoglobulin A antibody titres to a cytoplasmic protein extract of Candida albicans were determined by ELISA in saliva from 20 patients with oral candidosis and 21 controls. Patients had significantly increased levels of salivary IgA anti-Candida antibodies when compared with controls (P less than 0.001). Antibody levels were associated with IgA1 subclass in 90% of the patients; in contrast, IgA2 subclass was predominant in 67% of the controls. Antifungal therapy resulted in significantly decreased IgA1 titres (P less than 0.05) whilst the mean IgA2 antibody titre remained unchanged. The results indicate that Candida infection may change the subclass pattern of salivary IgA antibodies.

Expression of blood group H antigen type 2 chain by oral carcinoma cells after radiotherapy.

The blood group H antigen type 2 was investigated immunohistochemically in sections of 44 surgical specimens from the oral mucosa. These comprised 35 squamous cell carcinomas obtained from 22 patients and 9 specimens of clinically healthy mucosa. The carcinoma specimens included 10 primary lesions and 25 recurrent lesions from patients who had undergone radiotherapy. The results showed that the specimens of normal oral mucosa stained at higher antibody titers than either group of carcinomas, and that postradiation recurrent tumors stained at higher titers than primary tumors. In 10 patients, both preradiation and postradiation carcinomas were examined; the postradiation lesions showed increased reactivity in 5 patients, no change in 3 patients, and a decrease in antigen reactivity in 2 patients. The expression of antigen H type 2 in the recurrent tumors appeared to correlate with the estimated daily tumor radiation dose; tumors with specific antigen staining took twice as long to recur after radiotherapy than tumors without similar staining. The results suggest that the expression of the blood group H antigen type 2 substance, being a differentiation antigen, is enhanced by the effect of radiation on the malignant cell.

Expression of blood group H antigen by normal, benign, and carcinoma cells of the oral epithelium: immunohistochemical study using monoclonal antibody RS13.

The H antigen was investigated by an indirect immunoperoxidase method in sections of 65 surgical specimens from the oral mucosa. These comprised 29 squamous cell carcinomas, 28 benign lesions, and 8 specimens of clinically healthy mucosa. A monoclonal antibody (RS13) was used to identify the H antigen. The peroxidase stain was positive at high dilutions of the antibody in the epithelium of normal mucosa, and the titers were significantly higher than those within benign lesions (p less than 0.001) and carcinomas (p less than 0.001). However, the titers in the benign lesions varied considerably, with two specimens recorded as negative. In contrast, the reaction for the H antigen was negative in 19 specimens (66%) of the malignant lesions and the endpoint titers of the H positive carcinomas indicated marked loss of the antigen. This loss was significant when compared to the benign lesions (p less than 0.001). The results show that the loss of the H antigen on malignant epithelial cells may be a valuable marker for primary squamous cell carcinoma.

Serum IgG antibodies to lipopolysaccharides in various forms of periodontal disease in man.

Serum IgG antibody titres to lipopolysaccharides (LPS) from two strains of Actinobacillus actinomycetemcomitans were significantly elevated in juvenile periodontitis compared with other types of periodontal disease and with controls (p less than 0.05). The highest antibody titres to Bacteroides gingivalis LPS were in juvenile periodontitis, but this difference was significant only against the control group (p less than 0.01). In adult mild periodontitis there were higher antibody levels to LPS from Veillonella parvula compared with all other groups and controls (p less than 0.05). The possibility that high antibody titres to LPS from A. actinomycetemcomitans may play a protective role in juvenile periodontitis needs further investigation.

Relationship between the serum autoantibody titers and the clinical activity of pemphigus vulgaris.

With human tonsillar epithelium as a substrate for the indirect immunofluorescence assay, a statistically significant relationship was demonstrated between the clinical activity of the disease and the autoantibody titers in seventy-eight serum samples obtained from patients with pemphigus vulgaris. High pemphigus antibody titers were associated with severe ulceration, intermediate titers with moderate lesions, and low titers with mild pemphigus. However, the sequential studies revealed that this relationship was not always consistent throughout the course of the disease, and in three out of fourteen patients the correlation was either poor or negative. These results indicate that serial pemphigus antibody titers may not be consistent enough to be used reliably as the sole guide for monitoring disease activity.

Identification of pemphigus-like antigens expressed by SCaBER cells.

Indirect immunofluorescence revealed the presence of pemphigus-like antigenic activity on SCaBER cell monolayers after incubation with sera from patients with pemphigus vulgaris. In contrast, no fluorescence was observed after incubation of the cells with sera from patients with bullous pemphigoid, systemic lupus erythematosus or healthy individuals. In order to identify this pemphigus-like antigen, the SCaBER cell membrane proteins were solubilized in non-ionic detergent, separated according to their molecular weight by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and electrophoretically transferred to nitrocellulose sheets. After incubation with pemphigus serum followed by 125I-radiolabelled goat anti-human IgG, autoradiography revealed pemphigus antibody binding activity in the 105 Kd region in electrophoretograms of the unreduced SCaBER cell membrane extracts. This pemphigus-like antigen was isolated by affinity chromatography on the Sepharose-linked IgG fraction of sera from patients with pemphigus vulgaris.

Oral epithelial cells as the origin of pemphigus antigen in human saliva.

Absorption of pemphigus sera with normal human saliva or with scraped oral epithelial cells abolished the intercellular antibody titres, as revealed by indirect immunofluorescence. No pemphigus antigenic activity was detected on either human or macaque salivary glands, suggesting that the pemphigus antigen in saliva is a product of the epithelial lining of the oral mucosa. Indirect immunofluorescence on primary cultures of normal oral mucosa demonstrated that pemphigus antigen is synthesized by human oral epithelial cells. Salivary pemphigus antigen was isolated by affinity chromatography on Sepharose-linked IgG-fraction from pemphigus sera. Pemphigus antibody-binding material from the isolated fraction was detected in two bands with apparent molecular weights of 30,000 and 12,000 by immuno-blotting of reduced SDS-polyacrylamide gel electrophoretograms.

Expression of the CA1 determinant by carcinomas and by non-malignant epithelial cells in oral lesions.

The expression of the Ca antigen was investigated in 5 groups of oral lesions comprising 7 squamous cell carcinomas, 2 pre-invasive carcinomas, 7 lesions of types believed to predispose to carcinoma, 19 lesions of types that do not predispose to carcinoma and 5 biopsies of normal oral mucosa. Using an indirect immunoperoxidase method, the neoplastic epithelium reacted positively with the Ca1 antibody in only 4 out of 7 oral squamous cell carcinomas and the reaction varied between the specimens as to the intensity and number of positively stained cells. Several benign oral lesions specifically bound the Ca1 antibody in areas of epithelium showing infiltration with inflammatory cells. These lesions comprised 5 fibrous epulides, 1 pyogenic granuloma, 1 denture-induced hyperplasia and 1 non-diagnostic ulcer. We conclude that the Ca1 antibody is not sufficiently specific for the carcinoma to be of value in the diagnosis of malignant and premalignant lesions of the oral mucosa.

Topical acyclovir in the management of recurrent herpes labialis.

A double-blind, randomized, placebo-controlled trial of 5% acyclovir ointment was carried out for thirty-one episodes of herpes labialis in thirteen patients experiencing severe, frequent recurrences. In addition, these patients continued to use acyclovir ointment or placebo for the treatment of twenty-two further episodes. Treatment was initiated by the patients as soon as possible after onset of prodromal symptoms, and continued five times a day for 5 days. It was found that acyclovir ointment shortened the duration of lesions only by about one day, but it greatly increased the number of abortive lesions resulting from early application. Acyclovir ointment is well tolerated and appears to modify the course of severe recurrent herpes labialis when therapy is initiated by the patient during the prodrome.