Suppressor cell function in oral lichen planus.

Oral lichen planus (OLP) is a common inflammatory condition of the oral mucous membranes which affects between one and two percent of the general population. In accordance with the protracted clinical course of OLP and its association with known auto-immune diseases, the level of self-tolerance is questionable and possibly diminished in patients with this disorder. Normal suppressor T lymphocyte function is reputedly an essential element in the maintenance of self-tolerance, and deficient cell-mediated suppressor activity is implicated in the pathogenesis of auto-immune diseases. For assessment of in vitro cell-mediated suppressor activity in OLP, peripheral blood mononuclear cells (PBMC) from ten patients with OLP and from 11 control subjects were activated with the plant mitogen concanavalin A (Con A), followed by co-culture with autologous responder cells. The ability of irradiated Con A-activated cells to suppress the proliferation of Con A-stimulated responder cells was determined. Con A-induced suppressor activity of PBMC in the OLP patients was significantly less than that in control subjects (p = 0.001). Results of the present investigation complement previous in vitro findings which provided indirect evidence of deficient cell-mediated suppressor activity in OLP, particularly a decreased proportion of circulating CD4+CD45RA+ lymphocytes and reduced Con A-stimulated PBMC proliferation. The depressed Con A-induced suppressor activity of PBMC in the OLP patients provides direct evidence of deficient in vitro cell-mediated suppressor function in OLP, and suggests that defective cell-mediated suppressor circuits and reduced self-tolerance may be involved in the pathogenesis of this disorder.

P. gingivalis-specific T-cell lines produce Th1 and Th2 cytokines.

Cytokines produced by T-cells in periodontal lesions may determine the nature of the adaptive immune response. Since different antigen-presenting cells (APC) may direct the Th1/Th2 response, P. gingivalis-specific T-cell lines were established by different APC subpopulations, and their cytokine profiles were determined. Peripheral blood mononuclear cells induced similar percentages of IL-4+ and IFN-gamma+ T-cells and lower percentages of IL-10+ T-cells. Epstein-Barr virus-transformed B-cells (LCL) induced higher percentages of IL-4+ cells than IFN-gamma+ cells, with lower percentages of IL-10+ cells. Peripheral blood mononuclear cells induced a higher percent of IFN-gamma+ CD8 cells than LCL (p = 0.004). Purified B-cells, monocytes, and dendritic cells induced similar percentages of IL-4+ and IFN-gamma+ cells, although again, the percentage of IL-10+ cells was lower. The results of the present study have demonstrated that, as measured by FACS analysis of intracytoplasmic cytokines, P. gingivalis-specific T-cells produce both Th1 and Th2 cytokines, regardless of the APC population.

Cell-surface proteoglycan expression by lymphocytes from peripheral blood and gingiva in health and periodontal disease.

Cell-surface proteoglycans are involved in lymphocyte migration and activation. This study investigated the expression of syndecan-1, syndecan-4, and glypican in peripheral blood lymphocytes and by lymphocytes in variously inflamed periodontal tissues. Gingival specimens from healthy, gingivitis, or chronic periodontitis sites were stained by means of antibodies against B- and T-lymphocytes and also syndecan-1, syndecan-4, and glypican. Syndecan-1 expression by peripheral blood mononuclear cells (PBMC) from healthy, gingivitis, and chronic periodontitis subjects was assessed by flow cytometry. Syndecan-1 was expressed by B-cells/plasma cells but not T-cells in both gingivitis and chronic periodontitis lesions. Both B-cells/plasma cells and T-cells in gingivitis and chronic periodontitis expressed syndecan-4. Glypican was expressed only by macrophages. Stimulation of PBMC with mitogens and growth factors modulated syndecan-1 expression in both the T- and B-cells. Thus, cell-surface proteoglycan expression by lymphocytes in periodontal inflammation is cell-type-specific and may be modulated by inflammation.

Preliminary functional analysis of human epidermal T cells.

The function of human epidermal T cells (ETC) is unknown. In the present study, dermal T cells (DTC), ETC and keratinocytes were cultured from normal human skin. DTC and ETC lines were expanded in medium containing interleukin 2. The autologous keratinocytes were transfected with a human papillomavirus 16 E6 and E7 plasmid to produce an immortal keratinocyte line "HEK001". Lymphocyte migration and adhesion to HEK001 was assessed in calcein fluorimetric assays. ETC migrated towards HEK001 three to four times more than DTC. ETC adhered to HEK001 two to four times more than DTC. The proportion of ETC expressing the cutaneous lymphocyte-associated antigen was greater than that of DTC (26% and 1%, respectively). The keratinocyte line HEK001 expressed ICAM-1 following stimulation with TNF-alpha or IFN-gamma and following coculture with autologous cutaneous T cells. A blocking anti-ICAM-1 antibody reduced DTC and ETC adhesion to HEK001 by 30% and 50%, respectively. Therefore, cutaneous T cells may upregulate keratinocyte ICAM-1 expression which mediates adhesion to autologous keratinocytes. These results are consistent with the hypothesis that the ETC and DTC populations are distinct. Both directed migration (epidermotropism) and selective retention may be involved in the development and maintenance of the ETC population in normal human skin.

A quantitative cytological study of lesional and non-lesional mucosa in oral lichen planus.

Smears of buccal mucosa, dorsal surface of the tongue and floor of mouth were taken from 10 patients with histologically confirmed oral lichen planus and 12 healthy age- and sex-matched controls. In buccal smears, no significant differences in cytoplasmic and nuclear areas were observed between lesional, adjacent non-lesional and control tissues. However, the cytoplasmic area in smears from lichen planus lesions on the dorsum of the tongue and adjacent clinically normal mucosa was reduced compared with healthy controls. The cytoplasmic: nuclear ratio in smears from clinically normal floor of mouth in oral lichen planus was similarly reduced. Papanicolaou-stained smears from buccal lichen planus showed increased keratinization compared with normal buccal mucosa. These findings demonstrate that quantitative cytology can detect both cytoplasmic and nuclear changes in oral lichen planus.

Distribution of interleukin-2, -4, -10, tumour necrosis factor-alpha and transforming growth factor-beta mRNAs in oral lichen planus.

In the present study, MRNA for the cytokines interleukin-2 (IL-2), IL-4, IL-10 tumour necrosis factor-alpha (TNF-alpha) and transforming growth factor beta-1 (TGF-beta-1) were investigated in oral lichen planus (OLP) lesions using in situ hybridization with 35S-labelled oligonucleotide probes on frozen tissue sections. In addition, the expression of interferon-gamma (IFN-gamma), IL-10 and IL-4 mRNAs was analysed in cultured lesional T lymphocytes from oral lichen planus by polymerase chain reaction. Cells expressing mRNA for IL-2, IL-4, IL-10, TNF-alpha and TGF-beta 1 were found in all the biopsies studied. Approximately 1-2% of the total number of infiltrating cells in the lesions were positive for each of the different cytokine mRNAs. Most biopsies contained basement membrane-oriented, mRNA-positive cells. In the cultured T-cell lines, message for IFN-gamma was detected in all the patients, IL-10 in all but one, and IL-4 in just one of the seven patients investigated. The results suggest that mRNA for both pro- and anti-inflammatory cytokines, i.e., mixed T-helper 1 (TH1) and TH2 cytokine profiles, are generated simultaneously by a limited number of cells in chronic lesions of OLP.

Mucocele of the anterior lingual salivary glands (glands of Blandin and Nuhn): report of 5 cases.

The anterior lingual salivary glands (glands of Blandin and Nuhn) are mixed mucous and serous glands that are embedded within the musculature of the anterior tongue ventrum. Five cases of mucocele of the glands of Blandin and Nuhn are presented. These mucoceles on the anterior tongue ventrum were exophytic and resembled pyogenic granulomata, polyps, or squamous papillomata. In 2 cases, the onset of the mucocele was associated with trauma to the anterior tongue. All cases were mucus extravasation phenomena. A history of trauma and recovery of mucus with fine needle aspiration are helpful in the clinical diagnosis of mucocele of the glands of Blandin and Nuhn, as are the following characteristics of the mucocele: rapid onset, increase and reduction in size, bluish color, and fluid-filled consistency. During surgery, the glands that are deep in the tongue musculature are commonly left behind, resulting in persistence of the lesion. Careful clinical evaluation of these lesions and preoperative awareness of the surgical anatomy of the glands of Blandin and Nuhn may minimize the need for repeated surgical procedures.

Oral cancer in Australia: 1983-1996.

BACKGROUND: The aims of this study were to identify differences in oral cancer incidence and mortality between sexes, age groups, oral sites and Australian States and Territories and recent trends in oral cancer incidence, mortality and age-profile over time. METHODS: Data were obtained from the Australian Institute for Health and Welfare and were age-standardized to the Australian 1991 Population Standard. Differences and trends were assessed with the Wilcoxon matched-pairs signed-ranks test and the Spearman correlation test, respectively. RESULTS: In Australia in 1996, there were 2173 new oral cancers and 400 deaths due to oral cancer, the majority of oral cancers were in the 60+ age group, oral cancer affected men more than women (>2:1), lip cancer accounted for more than 50 per cent of oral cancers and the oral cancer mortality-to-incidence (M:I) ratio was greatest in ACT and NSW and least in QLD and SA. From 1983 to 1996, the annual incidence of lip cancer increased while the M:I ratio of lip cancer decreased. The annual incidence of cervical cancer decreased whereas the annual incidence of intra-oral cancer remained constant. The M:I ratio of cervical cancer was consistently lower than the M:I ratio of intra-oral cancer. CONCLUSIONS: Reducing exposure to environmental carcinogens, increasing public awareness and population screening may reduce the incidence and mortality of oral cancer in Australia.

Oral lichen planus: causes, diagnosis and management.

Oral lichen planus (OLP) is a chronic inflammatory disease of unknown etiology. In this paper we review the clinical and histological features of OLP, process of OLP diagnosis, causes of OLP, management of OLP patients and medical treatment of OLP lesions. Approximately 0.2 per cent OLP patients develop intra-oral carcinoma each year compared with approximately 0.005 per cent Australian adults. Possible mechanisms of increased oral cancer risk in OLP patients are presented. The aims of current OLP therapy are to eliminate mucosal erythema and ulceration, alleviate symptoms and reduce the risk of oral cancer. Patient education may improve the outcomes of OLP therapy and further reduce the risk of oral cancer in OLP patients. Although OLP may be diagnosed clinically, appropriate specialist referral is required for: (i) histological diagnosis; (ii) assessment of causative/exacerbating factors, associated diseases and oral cancer risk; (iii) patient education and management; (iv) medical treatment; and (v) long-term review and re-biopsy as required.

Exfoliative cytology in clinical oral pathology.

Exfoliative cytology is a rapid, non-invasive procedure for assessing dysplastic change within the oral epithelium. The indications for oral exfoliative cytology are reviewed and a technique for cell collection and smear examination is presented. The value of exfoliative cytology in oral cancer screening programmes as a public health measure is discussed.

Current concepts in oral cancer.

Over 750 new intra-oral squamous cell carcinomas are registered in Australia each year. In this article, the authors review the epidemiology, aetiology, genetics and spread of intra-oral squamous cell carcinoma. The mechanisms of field cancerization are discussed. The prevention of intra-oral squamous cell carcinoma is highlighted and future treatments are presented.

The high risk human papillomaviruses and oral cancer: evidence for and against a causal relationship.

Oncogenic human papillomaviruses (HPVs) have been detected in oral squamous cell carcinoma (SCC). HPV16 is the most frequently detected HPV type in oral SCC and is present in up to 22% of cases, either alone or in combination with other HPV types. HPV18 is present in up to 14% of cases. HPV16 and HPV18 are present together in approximately 6% of cases. However, HPV16 and 18 are also detected in normal oral mucosae (10% and 11% of subjects, respectively). These data suggest that high risk HPV infection may be a co-factor in oral carcinogenesis and that latent HPV infection of the oral mucosa is common. A role for HPV infection in oral carcinogenesis is supported by the ability of high risk HPVs to immortalize oral keratinocytes in vitro. Immortalization may involve (i) deactivation of pre-formed tumor-suppressor proteins by viral oncoproteins, (ii) blocking of tumor-suppressor gene transcription as a result of HPV oncogene insertion or (iii) stimulation of cellular oncogene transcription by the upstream insertion of HPV-derived transcription activating sequences. Hence, infection of oral keratinocytes with high risk HPV may be involved in the pathogenesis of some oral SCCs although the evidence implicating HPV in oral carcinogenesis is, at present, mainly circumstantial.

Phenotype and suppressor activity of T-lymphocyte clones extracted from lesions of oral lichen planus.

Lymphocytes were extracted from six biopsy specimens of oral lichen planus. T-lymphocyte lines were expanded in culture with phytohaemagglutinin and interleukin 2, and cloned by limiting dilution. Fifteen T-cell clones were isolated with a probability of clonality of 96.3%. The majority of clones (n = 13) expressed the alpha beta T-cell receptor, and of these, 11 were CD8+ and two were CD4+. Two clones were CD4- and CD8-, and expressed the gamma delta T-cell receptor. The ability of these clones (effectors) to suppress concanavalin-A-stimulated proliferation of autologous lesional T-cell lines (responders) was assessed. Maximum suppressor activity ranged from 17 to 100%. The majority of clones (n = 12), including a CD3+ CD4+ CD8-alpha beta+ clone, displayed suppressor activity which was proportional to the effector to responder ratio. A CD3+CD4+CD8-alpha beta+ clone and a CD3+CD4-CD8-gamma delta+ clone displayed substantial helper activity at higher effector to responder ratios. These results demonstrate differential helper and suppressor activity of T-lymphocyte clones extracted from oral lichen planus lesions. The balance between help and suppression may be a fundamental determinant of immunological activity within the lymphocytic infiltrate of oral lichen planus, and hence may dictate the clinical behaviour of the disease.

Autocytotoxic T-cell clones in lichen planus.

We examined the in vitro cytotoxic activity of cutaneous T-cell lines and clones from lichen planus (LP) patients against autologous epidermal keratinocytes. T cells were cultured from LP lesions and adjacent clinically normal skin and cloned by limiting dilution. Keratinocytes were cultured from LP lesions and adjacent clinically normal skin and immortalized by transfection with the E6 and E7 genes from human papillomavirus 16 (HPV16). The lesional T-cell line from one LP patient contained 27% gammadelta+ T cells and was significantly more cytotoxic against autologous lesional keratinocytes than the T-cell line from clinically normal skin. Clones isolated from the lesional T-cell line were significantly more cytotoxic against autologous lesional keratinocytes than clones isolated from the non-lesional T-cell line. Most cytotoxic clones from LP lesions were CD8+ and most non-cytotoxic clones from LP lesions were CD4+. One cytotoxic clone was CD4- and CD8- and expressed the gammadelta T-cell receptor. Two CD8+ LP lesional T-cell clones showed dose-dependent killing of HPV16 E6/E7-immortalized autologous lesional and normal keratinocytes, but no cytotoxic activity against Epstein-Barr virus-transformed autologous B-cell blasts. The cytotoxic activity of CD8+ lesional T-cell clones against autologous lesional keratinocytes was partially blocked with anti-major histocompatibility complex (MHC) class I monoclonal antibodies. These data support the hypothesis that CD8+ lesional T cells recognize an antigen associated with MHC class I on lesional keratinocytes and that CD8+ cytotoxic T cells lyse keratinocytes in LP lesions.

Review article: The role of oncogenes, tumour suppressor genes and growth factors in oral squamous cell carcinoma: a case of apoptosis versus proliferation.

Mutation, deactivation and disregulated expression of oncogenes and tumour-suppressor genes may be involved in the pathogenesis of oral squamous cell carcinoma (SCC). Deactivation of the p53 tumour-suppressor gene allows cell proliferation and blocks apoptosis of malignant oral keratinocytes. Mutation in the ras oncogene results in persistent mitogenic signalling. Upregulatioed c-Myc expression, in the presence of growth factors, provides an additional proliferative signal. Loss of retinoblastoma tumour-suppressor gene (Rb) function may contribute to oral keratinocyte hyperproliferation and recent evidence suggests that simultaneous deactivation of both p53 and Rb is required for tumourigenesis. Enhanced Bcl-2 and reduced Fas expression inhibit tumour cell apoptosis and may convey resistance to cytotoxic drugs and T cell-mediated cytotoxicity, respectively. Exogenous mutagens such as tobacco, alcohol and viral oncogenes may cause altered expression of oncogenes and tumour-suppressor genes in some cases of oral SCC. The impact of these mechanisms on future therapies for oral SCC is highlighted.

Clonal expansion of lymphocytes from oral lichen planus lesions.

Lymphocytes were extracted from 11 biopsy specimens of oral lichen planus (OLP) by collagenase digestion, and cell lines were expanded with repetitive cycles of stimulation (with phytohaemagglutinin) and rest in media supplemented with interleukin 2. Four OLP lines contained a majority of CD3+CD4-CD8+ cells, in six lines the CD4:CD8 ratio was between 1 and 2, and in one line the CD4:CD8 ratio was 5:1. Limiting dilution of nine lines at 0.3 and 1.0 cells/well resulted in viable wells (putative clones) with plating efficiencies ranging from 0.0 to 18.1 percent and 0.0 to 22.2 percent respectively. The majority of clones were CD3+CD4-CD8+alpha beta+gamma delta-, although three clones were CD3+CD4+CD8-alpha beta+gamma delta- and one clone was CD3+CD4-CD8- and expressed the gamma delta T cell receptor. T cell clones derived from lymphocytes extracted from OLP lesions may be generated and maintained in culture providing opportunity for their further phenotypic and functional characterisation. This strategy may facilitate the identification of a putative oral lichen planus-specific antigen and indicate the frequency of lichen planus-specific T cells within lesions of OLP.

[The changes of suppressor T-cells function in patients with RAU].

OBJECTIVE: To study the change of suppressor T-cell function in patients with RAU. METHODS: Samples of 12 patients with RAU active phase were studied by it suppressor assay. RESULTS: The T-cell suppress rates at ConA 1,2,4 and 8 mg/L were 60%, 40%, 27% and 20% respectively, and were evidently lower when compared with normal controls' 72%, 56%, 41% and 34% (P < 0.05). CONCLUSION: The suppressor T-cell function may be depressed in RAU patients.