Some specific human leukocyte antigen (HLA)-DR/DQ haplotypes are more important than individual HLA-DR and -DQ phenotypes for the development of mucocutaneous type of Behçet's disease and for disease shift from recurrent aphthous stomatitis to mucocutaneous type of Behçet's disease

Abstract: The phenotype and haplotype frequencies of human leukocyte antigens (HLA)‐DR and ‐DQ in 32 Chinese patients with the mucocutaneous (MC) type of Behçet’s disease (BD) were calculated and compared with those in 310 healthy control Chinese and with those in 80 Chinese patients with recurrent aphthous stomatitis (RAS). We found that the phenotype frequency of HLA‐DRw8 [corrected P (P_c)<0.005] and the haplotype frequencies of HLA‐DRw8/DQw1 (P_c<0.005), ‐DRw8/DQw5(w1) (P_c<0.0005), ‐DRw12(5)/DQw1 (P_c<0.005), ‐DRw12(5)/DQw6(w1) (P_c< 0.0005), and ‐DRw52/DQw1 (P_c<0.005) in patients with the MC type of BD were significantly greater than those in healthy control subjects. This finding suggests that individual Chinese with HLA‐DRw8 antigen and HLA‐DRw8/DQw1, ‐DRw8/DQw5(w1), ‐DRw12(5)/DQw1, ‐DRw12(5)/DQw6(w1) and ‐DRw52/DQw1 haplotypes are more likely to have the MC type of BD. Furthermore, the relative risks (RRs) of HLA–DRw8/DQw1 (5.6), ‐DRw8/DQw5 (w1) (10.0), and ‐DRw12(5)/DQw6(w1) (14.4) haplotypes in patients with the MC type of BD were equal to or higher than the RR of HLA‐DRw8 phenotype (5.6), suggesting that some of the HLA‐DR/DQ haplotypes may play more important roles than the individual HLA‐DR and ‐DQ phenotypes for the development of the MC type of BD. The phenotype frequencies of HLA‐DR5 (P_c<0.01), ‐DRw8 (P_c<0.005) and ‐DQw1 (P_c<0.05) as well as the haplotype frequencies of HLA‐DR5/DQw1 (P_c<0.005) and ‐DRw8/DQw1 (P_c<0.00005) in patients with the MC type of BD were significantly higher than those in patients with RAS. Moreover, the RRs of HLA‐DR5/DQw1 (29.1) and ‐DRw8/DQw1 (47.4) haplotypes were greater than the RRs of HLA‐DR5 (10.4), ‐DRw8 (23.4) and ‐DQw1 (4.0) antigens. These results suggest that some specific HLA‐DR/DQ haplotypes may be more important than the individual HLA‐DR and ‐DQ phenotypes in the disease shift from RAS to the MC type of BD.     

HLA-DR and DQ antigens in Chinese patients with oral lichen planus

A study of HLA-DR and DQ typing in 44 patients with oral lichen planus (OLP) (28 women and 16 men) and in 107 normal controls of both sexes was performed by using Terasaki's oriental tray. Twenty-eight patients had erosive forms of lichen planus (OLPe). Serologic typing revealed a highly significant increase of HLA-DR 9 and Te 22 antigens in the patient group. Considering our findings and those in other autoimmune diseases, it is hypothesized that OLP is a localized autoimmune disease and the HLA-DR9 in Chinese takes the place of HLA-DR3 in Caucasians in carrying the genes which predispose to the development of autoimmune disease.     

Recurrent aphthous stomatitis in Behçet's disease: clinical featueres and correlation with systemic disease expression and severity

Behçet's disease (BD) is a multisystem disease, in which recurrent aphthous stomatitis (RAS) is a universal finding. We studied the expression of RAS in patients with BD, and the correlation between major or minor RAS and systemic expression and severity of the disease. Thirty-five patients with BD were studied, of whom 13 (37%) had major, 21 (60%) had minor and one (3%) had herpetiform RAS. The frequency of major RAS was significantly higher compared with a control group of patients with idiopathic RAS (37% vs 9%, P<0.05). The BD patients with major RAS had significantly more relapses of oral ulceration in a year, higher numbers of oral ulcers per relapse, and longer duration of aphthous episodes, compared with patients with minor RAS. Oral ulcers also appeared at a significantly younger age in patients with major than with minor RAS. However, the systemic expression of the disease, as well as the disease severity score, were similar in patients with major and minor RAS. The results of this study indicate that major RAS is common in patients with BD, and is associated with a more severe, repeated and prolonged oral disease. Nevertheless, the presence of major RAS in BD does not predict a more severe systemic illness.     

Human cytomegalovirus as a potential etiologic agent in recurrent aphthous ulcers and Behçet's disease

In a preliminary study on anti-human cytomegalovirus (HCMV) antibody (Ab) by ELISA. the serum anti-HCMV/IgG Ab concentrations in 22 patients with recurrent aphthous ulcers (RAU) in the remission stage were found to be significantly higher than in 22 control subjects (121±42 vs 100±27, P<0.05) and in 39 patients with RAU in the active stage (121±42 vs 88±45, P<0.01). Therefore, the potential of HCMV as an etiologic agent in RAU was proposed and studies using the polymerase chain reaction (PCR) and in situ hybridization (ISH) have been performed to investigate the possible presence of HCMV DNA in pre-ulcerative oral aphthous lesions in patients with RAU or Behçet's disease (BD) of the mucocutaneous type. For this purpose, formalin-fixed biopsy specimens were obtained from 13 pre-ulcerative oral aphthous lesions, 2 samples of normal oral mucosa and 1 ileal mucosal lesion from 9 RAU patients and 4 BD patients. Five specimens of normal oral mucosa from 5 normal control subjects and 12 specimens of oral erosive or ulcerative lesions from 12 patients with erosive lichen planus (ELP) were also included. By PCR, HCMV DNA was detected in 5 of the 13 (38.5%) pre-ulcerative oral aphthous lesions. 3 from RAU patients and 2 from BD patients. The ileal mucosa specimen was also HCMV DNA-positive, whereas HCMV DNA was not demonstrated in any of the 7 specimens of normal oral mucosa from RAU patients and normal control subjects; 12 specimens of oral lesions from ELP patients were similarly negative. ISH did not detect HCMV DNA in any of the biopsy specimens from RAU patients and control subjects. Our findings suggest that HCMV may be an etiologic agent in some cases of RAU and BD.     

Clinical and immunological effects of azithromycin in Behçet''s disease

Background: The aim of this study was to evaluate the effects of azithromycin on mucocutaneous manifestations, oral health and immune response in Behçet's disease (BD). Methods: Eight BD patients with active mucocutaneous symptoms were treated with azithromycin for 4 weeks. Oral health, clinical manifestations and in vitro interleukin (IL)‐12, interferon (IFN)‐γ, IL‐10 and monocyte chemotactic protein (MCP)‐1 responses were evaluated before and after treatment. Results: The number of folliculitic lesions, healing time of oral ulcers and scores of plaque indexes (PLIs) were lower after azithromycin treatment (P < 0.05). Scores of PLIs correlated positively with the healing time of oral ulcers (P = 0.02). Although a trend towards increased stimulated IL‐10 responses with azithromycin was observed, no statistically significant difference was found. Stimulated and unstimulated MCP‐1, IFN‐γ and IL‐12 responses were similar before and after treatment (P > 0.05). Conclusion: Azithromycin was observed to be effective in decreasing folliculitic lesions and fastening the healing time of oral ulcers in BD.     

Relationship between periodontal parameters and Behçet's disease and evaluation of different treatments for oral recurrent aphthous stomatitis

Background and Objective:  The highest prevalence of Behçet's disease (BD) has been reported in Turkey, and therefore research of relations between BD and other disorders have become important in the Turkish population. Since oral aphthous ulcers impair tooth brushing, reducing complaints about aphthous ulcers will motivate the patient to maintain better oral hygiene performance and will thus reduce plaque accumulation and periodontal scores. The purpose of this controlled case study was to evaluate the relationship between the severity of periodontal scores and Behçet's disease, and to compare the treatment modalities with neodymium-doped yttrium aluminium garnet (Nd:YAG) laser and medication on the recurrent aphthous ulcers in BD patients by considering the degree of pre- and post-treatment pain, discomfort and functional complications.
Material and Methods:  The periodontal status of 28 BD patients was evaluated according to periodontal indices. The BD patients were also assessed for clinical severity score as described previously. Levels of pre- and post-treatment pain and functional complications were assessed at patient visits on days 1, 4 and 7.
Results:  Periodontal indices, the number of oral ulcers and the daily frequency of tooth brushing were related to the severity scores of BD ( p  < 0.001). The results indicated that BD patients treated with the Nd:YAG laser had less post-treatment pain and fewer functional complications and reported immediate relief of pain and faster healing ( p  < 0.001).
Conclusion:  Our results suggest that periodontal status is worse in BD patients and is associated with disease severity; also, the Nd:YAG laser has better patient acceptance, shorter treatment time and lower rates of pain and post-treatment adverse events among BD patients with oral recurrent aphthous stomatitis.     

Preliminary evidence for an association of Epstein-Barr virus with pre-ulcerative oral lesions in patients with recurrent aphthous ulcers or Behiçet's disease

In this study we used the polymerase chain reaction (PCR), slot blot and Southern blot hybridization, direct sequencing and in situ hybridization (ISH) to show the possible presence of EBV-DNA in pre-ulcerative oral aphthous lesions of patients with recurrent aphthous ulcers (RAU) or Behet's disease (BD). For this purpose, formalin-fixed biopsy specimens were obtained from 13 pre-ulcerative oral aphthous lesions of nine RAU and four BD patients. Five specimens of normal oral mucosa (NOM) from five normal control subjects and 10 specimens of oral erosive or ulcerative lesions from 10 patients with erosive lichen planus (ELP) were also included. EBV-DNA was detected by PCR in 5 of the 13 (38.5%) pre-ulcerative oral aphthous lesions, two from RAU patients and three from BD patients. However, no EBV-DNA was demonstrated in five NOM specimens from normal control subjects and in 10 specimens of oral lesions from ELP patients. EBV-DNA was also demonstrated in patients'peripheral blood lymphocytes and/ or plasma, suggesting that the lymphocytes may be the reservoir of latent EBV infection and there is EBV shedding in the plasma. EBV-DNA was detected by ISH in only one PCR-positive case; the reaction product was found to deposit on the nuclei of some of the epithelial cells and lymphocytes. By immunohistochemistry, expression of Epstein-Barr nuclear antigen and EBV/C3d receptors was also noted in some of the epithelial cells and lymphocytes in this ISH-positive case. Therefore, we suggest that the epithelial cells of pre-ulcerative oral aphthous lesions may be infected by EBV through EBV-infected lymphocytes; also, the cytotoxic T lymphocyte-induced lysis of the EBV-infected epithelial cells, but not the virus-induced cytolysis, may be the main mechanism causing oral ulcer formation. Our data provide preliminary evidence for an association of EBV with pre-ulcerative oral aphthous lesions in RAU and BD patients.     

Expression of interleukin-2 receptor and HLA-DR on lymphocyte subsets of gingival crevicular fluid in patients with periodontitis

Expression of interleukin-2 receptor (IL2R) and HLA-DR on lymphocytes of gingival crevicular fluid (GCF) was examined by two-color flow cytometric analysis. GCF from 15 patients with periodontitis was collected by crevicular washing. Mononuclear cells were isolated by Ficoll-paque gradient centrifugation from inflamed gingival tissue (GT) and peripheral blood (PB) sampled from each of the 15 patients. Lymphocyte subsets were detected by using monoclonal antibodies (mAb) of Leu 12 (CD19), Leu 4 (CD3), Leu 3a (CD4) and Leu 2a (CD8) directed to B cells, T cells, helper/inducer T cells (Th) and suppressor/cytotoxic T cells (Ts), respectively. Anti-IL2R (CD25) and anti-HLA-DR were used as lymphocyte activation markers. IL2R- or HLA-DR-positive fractions in Th, Ts and B cells were calculated. Percentage of IL2R-positive fraction in Th (IL2R+ Th) of GCF (34.0%) was significantly higher than those of GT (18.4%) and PB (13.7%). IL2R-positive fraction in B cells (IL2R+ B) of GCF was the highest among the three groups (23.9% in GCF, 12.5% in GT, 6.3% in PB). Ts did not express IL2R regardless of the origin of the samples. Compared with PB and GT, GCF showed significantly higher HLA-DR expression on Th and Ts in GCF (PB: 8.7% and 27.1%; GT: 27.9% and 50.3%; GCF: 44.7% and 65.3%). These results suggest that lymphocytes in GCF were highly activated and are related to the local host immune response in periodontitis.     

口腔扁平苔藓与HLA的相关性研究

目的：探讨口腔扁平苔藓与人类白细胞抗原的相关性。方法：应用微量淋巴细胞毒试验,检测１９例上海汉族人口腔扁平苔藓外周血的人类白细胞抗原（ＨＬＡ－Ａ、－Ｂ、－ＤＲ、－ＤＱ）的抗原频率。结果：患者组ＨＬＡ－ＤＲ１抗原频率（３６．８４％）较正常对照组（１．０８％）显著增高（Ｐ＜０．００１）。结论：ＨＬＡ－ＤＲ１与口腔扁平苔藓明显相关。带有ＨＬＡ－ＤＲｌ相关基因的个体对口腔扁平苔藓具有遗传易感性     

Langerhans cells expressing HLA-DQ, HLA-DR and T6 antigens in normal oral mucosa and lichen planus

We compared Langerhans cells (LC) expressing HLA-DQ, HLA-DR and T6 antigens in biopsies from the same oral mucosal site in 12 patients with oral lichen planus and eight healthy volunteers. LC expressing each antigen were observed in all the specimens, but in lichen planus the cells were located in higher levels of the epithelium than in controls. Compared with controls, lichen planus contained significantly more HLA-DQ-positive LC (P = 0.04) and fewer HLA-DR-positive LC (P = 0.05), but there was no such difference in T6-positive LC. In lichen planus specimens, there were significantly more LC expressing HLA-DQ and T6 than HLA-DR (P = 0.0001 and 0.02 respectively); no such differences were found in normal mucosa. Epithelial cells in lichen planus expressed HLA-DR antigen, but not HLA-DQ or T6 antigens. We conclude that in lichen planus there is modulation of HLA-DR and HLA-DQ antigen expression by LC, or differences in the number of LC expressing those antigens.     

Expression of MHC Class II antigens (HLA DR, DP and DQ) by keratinocytes in oral lichen planus

The incidence and distribution of keratinocytes expressing the major histocompatability complex antigens HLA DR, DP and DQ and their relationship to the density of the inflammatory infiltrate was determined in lesions of oral lichen planus and normal mucosa using an immunoalkaline phosphatase technique. Seven of eight biopsies of lichen planus showed evidence of keratinocyte HLA DR expression but the pattern was variable both within and between biopsies. A significant increase in density of lymphocytes was found beneath those areas showing HLA DR expression throughout the prickle cell layer compared with those areas showing patchy or no expression. No evidence for keratinocyte expression of HLA DP or DQ was found. These results suggest that expression of HLA DR by keratinocytes in lichen planus may be induced by the lymphocytic infiltrate perhaps as a result of interferon gamma production. The functional significance of such MHC Class II expression remains unknown.     

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.     

Epithelial HLA-DR expression in labial salivary glands in Sjögren''s syndrome and nonspecific sialadenitis

The expression of the Class II major histocompatibility antigen HLA-DR was quantified in the epithelial cells of labial salivary glands from patients with Sjögrens Syndrome (SS) and compared with similar expression in glands showing non-specific sialadenitis and normal controls. In all glands more duct cells were positive than acinar cells but only in sialadenitis and SS was strong epithelial staining seen. The proportions of duct and acinar cells expressing HLA-DR were increased between normals and sialadenitis (P < 0.01) and between sialadenitis and SS (P < 0.001). However, for all cases increased expression of HLA-DR correlated to the increased proportion of inflammatory cells in the gland (P < 0.01). The results indicate that although HLA-DR is expressed on the epithelial cells in the glandular lesions of SS, this is not specific as it is also seen in sialadenitis. This supports the view that such expression is secondary to an inflammatory infiltrate and may not be of importance in initiating autoimmune tissue damage.     

CD9 and HLA-DR expression by crevicular epithelial cells and polymorphonuclear neutrophils in periodontal disease

BACKGROUND, AIMS: The composition of gingival crevicular fluid (GCF) is likely to reflect inflammatory modifications that take place in the gingiva during periodontal diseases. METHOD: In this study, GCF was collected at 3 different sites from 23 periodontal patients. The sites were assessed to be healthy, presenting gingivitis or periodontitis. 10 healthy individuals without any form of periodontal disease formed the control group and were sampled at one site each. The cell content of GCF was collected using Durapore Millipore strips, and 2 types of cells were studied: epithelial cells (EC) and polymorphonuclear neutrophils (PMN). The expression of CD9 and HLA-DR within or on the surface of these cells was studied in immunofluorescence on cytospin smears. RESULTS: Both CD9 and HLA-DR expression on EC differed significantly from control subjects, and the latter decreased according to the severity of the pathology. None of the PMN found in controls expressed CD9 or HLA-DR. However, in periodontal patients, the expression of HLA-DR within PMNs was detectable and increased according to the severity of lesions. CD9 expression on PMNs also increased with inflammation. CONCLUSION: This study shows that clinically healthy sites of periodontal patients already present signs of immunological activation characterised by a down modulation of HLA-DR expression on EC and an upregulation of these 2 molecules in PMN.     

Increased risks of dental caries and periodontal disease in Chinese patients with inflammatory bowel disease

Objectives: Inflammatory bowel disease (IBD) may be associated with oral diseases, but few relevant studies have been reported in China. This study aimed to compare the prevalence, severity and extent of dental caries and periodontal disease in Chinese IBD patients and healthy controls. Materials and methods: In this cross-sectional study, questionnaires and oral examinations were completed for 389 IBD patients [265 with Crohn’s disease (CD) and 124 with ulcerative colitis (UC)] and 265 healthy controls based on the established criteria of the World Health Organization. Tobit regression, multiple linear regression and logistic regression were performed to analyse the data. Results: After adjusting for confounders, the decayed, missing and filled surfaces indices were significantly increased in the CD and UC patients compared with those in the controls (P < 0.001). Patients with CD [odds ratio (OR) = 4.27, 95% confidence interval (95% CI): 2.63–6.95, P < 0.001] and UC (OR = 2.21, 95% CI: 1.24–3.94, P = 0.007) had significantly higher risks of dental caries than controls. Significantly higher percentages of sites with probing pocket depth ≥ 5 mm and clinical attachment loss ≥ 4 mm were observed in CD and UC patients compared with controls (P < 0.001). A fully adjusted model revealed that CD and UC were risk indicators for periodontitis (OR = 4.46, 95% CI: 2.50–7.95, P < 0.001; OR = 4.66, 95% CI: 2.49–8.71, P < 0.001, respectively). No significant differences in dental caries and periodontal disease were observed between the CD and UC patients. Conclusions: Chinese IBD patients have a higher prevalence, severity and extent of dental caries and/or periodontal disease than controls, and require oral health education and multidisciplinary treatment.Key words: Dental caries, periodontal disease, inflammatory bowel disease, Crohn’s disease, ulcerative colitis     

牙周病的中医药治疗概况和展望

牙周病的中医药治疗在我国有悠久的历史，它与牙周基础治疗结合，可以弥补西医在牙周病治疗上偏重局部的缺陷，获得更好的治疗效果。本文主要从以下几个方面进行综述：①中医药治疗牙周病的进展情况，②中西医结合治疗牙周病的合理性，③中医药治疗牙周病研究方面的不足及展望。     

Epithelial expression of HLA class II antigens and Feγ receptors in patients with adult periodontitis

Abstract The distribution of HLA class II (DR, DP, DQ) and FcγR (I, II, III) was analyzed in the epithelia of patients with advanced marginal periodontitis using cryostat sections incubated with monoclonal antibodies (MoAb) against the Langerhans cell (LC) (CDla) and various subtypes of HLA class II and FcγR, and the indirect immunofluorescence technique. In the oral gingival epithelium (OGE), LC were concentrated subjacent to the connective tissue papillae, while in the pocket epithelium (PE), they were most abundant at the gingival margin. HLA-DP, DQ, and DR stained LC in both OGE and PE. HLA-DQ+ LC were significantly fewer than DP+ and DR+ LC. HLA-DR also stained keratinocytes (KC) in the whole extension of both OGE and PE. HLA-DP was also observed on KC, but not HLA-DQ. FcγR II stained both LC and focal areas of KC. In PE FCγR II+ LC were concentrated near the bottom of the pocket, while in the OGE, they were concentrated at the gingival margin. FcγR III was present only on KC, especially in the basal and suprabasal layer. The results indicate that the epithelial cells are actively involved in the development and maintenance of the inflammation of periodontal disease.