Extent and diversity of inflammatory cell infiltrates in squamous cell carcinomas and basal cell epitheliomas of the head and neck

Using monoclonal antibodies reactive with Langerhans' cells (LCs), macrophages, and T cell subpopulations, the density and proportions of cells of the immune system of the normal oral mucosa were determined immunohistochemically, and compared with findings in oral squamous cell carcinomas (SCC) and basal cell epitheliomas (BCE). In normal oral epithelia, the dominant cell type was the LC, positive for CD 1, and expressing HLA-DR antigens (DR⁺). Many intraepithelial cells were lymphocytes of the suppressor/cytotoxic phenotype (CD 8⁺), which was also the most prominent cell type in the normal mueosal slroma. Significant differences were observed for the content of CD 8-, OKM 1-, and CD 4-positive cells in the epithelium of normal oral mucosa, SCC, and BCE, and for the amount of CD 1-positive Langerhans cells in the connective tissue of the different groups of tissues. When CD 4/CD 8 ratios were calculated, differences between SCC and BCE became most evident. A CD 4/CD 8 ratio greater 0.5 was seen to be characteristic for BCE. Thus, in contrast to the striking preponderance of suppressor/cytotoxic lymphocytes (CD 8⁺) in SCC, BCE showed typically almost balanced numbers of suppressor/cytotoxic (CD 8⁺) and helper/induced (CD 4⁺) lymphocytes. This finding further underlines the biological differences recognized between these most common neoplasias of the head and neck.     

Pattern of distribution of T lymphocytes, Langerhans cells and HLA-DR bearing cells in normal human oral mucosa

The tissue distribution of helper/inducer and suppressor/cytotoxic T cells, Langerhans cells (LC) and HLA-DR bearing cells was determined in normal oral mucosa by use of monoclonal antibodies OKT4, OKT8, OKT6 and OKIa1, respectively. OKT4+ and OKT8+ cells were invariably present in normal oral epithelium and in the lamina propria. OKT8+ cells were consistently seen inside the basal cell layer of the epithelium. The distribution of LC in oral epithelium showed regional variation. In palatal epithelium LC were evenly distributed in the basal half of the epithelium, whereas in buccal mucosa the highest concentration of LC was seen in the epithelium overlying the tips of connective tissue papillae. OKIa1 stained dendritic cells in the epithelium and plump cells with small dendritic processes in the connective tissue. Some of the latter were located close to the basal cells of the epithelium. The consistent relationship between immunocompetent cells and the epithelium of the oral mucosa suggests the presence of a local immunologic defence barrier in the oral mucosa.     

Pattern of distribution of T lymphocytes, Langerhans cells and HLA-DR bearing cells in normal human oral mucosa

The tissue distribution of helper/inducer and suppressor/cytotoxic T cells, Langerhans cells (LC) and HLA-DR bearing cells was determined in normal oral mucosa by use of monoclonal antibodies OKT4, OKT8, OK.T6 and OKIal, respectively. OKT4+ and OKT8+ cells were invariably present in normal oral epithelium and in the lamina propria. OKT8+ cells were consistently seen inside the basal cell layer of the epithelium. The distribution of LC in oral epithelium showed regional variation. In palatal epithelium LC were evenly distributed in the basal half of the epithelium, whereas in buccal mucosa the highest concentration of LC was seen in the epithelium overlying the tips of connective tissue papillae. OKIal stained dendritic cells in the epithelium and plump cells with small dendritic processes in the connective tissue. Some of the latter were located close to the basal cells of the epithelium. The consistent relationship between immunocompetent cells and the epithelium of the oral mucosa suggests the presence of a local immunologic defence barrier in the oral mucosa.     

Extent and diversity of inflammatory cell infiltrates in squamous cell carcinomas and basal cell epitheliomas of the head and neck

Using monoclonal antibodies reactive with Langerhans' cells (LCs), macrophages, and T cell subpopulations, the density and proportions of cells of the immune system of the normal oral mucosa were determined immunohistochemically, and compared with findings in oral squamous cell carcinomas (SCC) and basal cell epitheliomas (BCE). In normal oral epithelia, the dominant cell type was the LC, positive for CD 1, and expressing HLA-DR antigens (DR+). Many intraepithelial cells were lymphocytes of the suppressor/cytotoxic phenotype (CD 8+), which was also the most prominent cell type in the normal mucosal stroma. Significant differences were observed for the content of CD 8-, OKM 1-, and CD 4-positive cells in the epithelium of normal oral mucosa, SCC, and BCE, and for the amount of CD 1-positive Langerhans cells in the connective tissue of the different groups of tissues. When CD 4/CD 8 ratios were calculated, differences between SCC and BCE became most evident. A CD 4/CD 8 ratio greater 0.5 was seen to be characteristic for BCE. Thus, in contrast to the striking preponderance of suppressor/cytotoxic lymphocytes (CD 8+) in SCC, BCE showed typically almost balanced numbers of suppressor/cytotoxic (CD 8+) and helper/inducer (CD 4+) lymphocytes. This finding further underlines the biological differences recognized between these most common neoplasias of the head and neck.     

Immunohistochemical demonstration of T cell subsets and accessory cells in oral lichen planus

The nature and distribution of mononuclear cells in 30 non-ulcerated lesions of oral lichen planus (OLP) was investigated, using an immunoperoxidase technique. Most of the infiltrating cells consisted of a mixture of Leu 2a+ and Leu 3a+/3b+ T cells present in the stroma. This study proved histometrically that the emigration of lymphocytes through subepithelial vessels was not selective for major subsets of T cells and subsequent migration to the epithelium was predominant in suppressor/cytotoxic T cell infiltration. HLA-DR+/DQ+ monocyte/macrophage and Langerhans cells formed a relatively minor component of the cellular infiltrates, whereas a considerable number of T cells expressed the MHC antigens. Also, the keratinocytes of the epithelium expressed only DR antigens. These results support the concept that LP is associated with lymphokine-generated inflammation induced by helper/inducer T cells or activated T cells which would include direct basal cell damage or local immunosuppression by suppressor/cytotoxic T cells. Furthermore, this study suggests that monocytes/macrophages and Langerhans cells played a role in antigen presentation, and also that keratinocytes may possess a similar function.     

Immunohistochemical demonstration of T cell subsets and accessory cells in oral lichen planus

The nature and distribution of mononuclear cells in 30 non-ulcerated lesions of oral lichen planus (OLP) was investigated, using an immunoperoxidase technique. Most of the infiltrating cells consisted of a mixture of Leu 2a+ and Leu 3a+/3b+ T cells present in the stroma. This study proved histometrically that the emigration of lymphocytes through subepithelial vessels was not selective for major subsets of T cells and subsequent migration to the epithelium was predominant in suppressor/cytotoxic T cell infiltration. HLA-DR+/DQ+ monocyte/macrophage and Langerhans cells formed a relatively minor component of the cellular infiltrates, whereas a considerable number of T cells expressed the MHC antigens. Also, the keratinocytes of the epithelium expressed only DR antigens. These results support the concept that LP is associated with lymphokine-generated inflammation induced by helper/inducer T cells or activated T cells which would include direct basal cell damage or local immunosuppression by suppressor/cytotoxic T cells. Furthermore, this study suggests that monocytes/macrophages and Langerhans cells played a role in antigen presentation, and also that keratinocytes may possess a similar function.     

髓样来源的抑制细胞在舌癌小鼠中的表达及意义

目的 评价舌癌小鼠外周血和病变部位髓样来源的抑制细胞（MDSC）的变化及意义。方法 建立4NQO舌癌小鼠模型,观察MDSC细胞在外周血的表达,同时观察T细胞亚群的表达,评价MDSC细胞与T细胞亚群及CD4⁺/ CD8⁺比例变化的相关性。观察MDSC细胞在舌黏膜病变部位的表达,并检测舌组织精氨酸酶1（ARG-1）mRNA的表达。结果 4NQO小鼠外周血中MDSC比例随着肿瘤进展显著升高（P<0.01）,外周血中MDSC比例与相应CD3⁺CD8⁺T细胞的比例呈正相关,与CD4⁺/CD8⁺比呈负相关关系。4NQO小鼠异常增生区域的MDSC细胞较正常对照组有所增加,在舌癌组织内及与正常组织的交界处,均浸润了大量的MDSC细胞,而且ARG-1 mRNA水平显著升高（P<0.01）。结论 MDSC细胞在舌癌组织和外周血中的扩增可能是肿瘤进展的重要原因。MDSC细胞可能成为口腔癌免疫治疗的潜在靶点。     

Immunohistochemical study of oral keratoses including lichen planus

Biopsies of non-ulcerated oral mucosa from 13 patients with oral lichen planus and 12 patients with leukoplakia were immunohistochemically stained using monoclonal antibodies to pan T, pan B, T helper and T suppressor/cytotoxic cells and the stained lymphocytes enumerated using an image analyser. The results show the preponderance of T cells infiltrating both oral lichen planus and leukoplakia. The T helper: T suppressor/cytotoxic cell ratio was the same (1:2) for both oral lichen planus and leukoplakia. A similar proportion of T suppressor/cytotoxic cells was found infiltrating the epithelium. These data indicate that T cell subset analysis is of no value in distinguishing oral lichen planus from other oral keratoses.     

46例HIV/AIDS患者在高效抗逆转录病毒治疗第一年间口腔念珠菌感染及免疫状况动态变化

目的:监测HIV/AIDS患者在高效抗逆转录病毒治疗(HAART)第一年间口腔念珠菌感染及免疫状态变化,并探讨其间的关系.方法:随访46例HIV/AIDS患者,在HAART基线、治疗后3、6、12个月检测患者CD4+T淋巴细胞计数,对患者进行口腔检查并记录口腔念珠菌病发生情况,采集患者口腔含漱液,使用沙保罗琼脂培养基和...     

Enumeration of T cell subsets with monoclonal antibodies in minor salivary glands of patients with rheumatoid arthritis

Labial salivary glands of 51 patients with rheumatoid arthritis and those of 25 control patients were examined by the ANAE (acid alpha-naphthyl acetate esterase) technique to determine the percentages of B- and T-lymphocytes and mononuclear phagocytes (MPS cells). Using monoclonal antibodies (OKT3 for all T cells, OKT4 for helper/inducer T cells, OKT6 for thymocytes, and OKT8 for suppressor/cytotoxic T cells) T cell subsets were enumerated. B-lymphocytes predominated in both series of salivary glands, and the percentages of B and T cells were equal in both series. The absolute cell counts in the salivary glands of rheumatics were significantly higher (P less than 0.001) than in those of healthy controls. The number of OKT4+ cells was increased in rheumatics, leading to an elevated OKT4+/OKT8+ ratio when compared with that in controls (P less than 0.01). The results suggest that the basic phenomenon behind the B cell hyperactivity noticed in rheumatics might be due to increased activity of T helper cells rather than reduced number of T suppressor cells, which were shown to remain almost unaffected in the salivary glands of patients with rheumatoid arthritis.     

Intra-epithelial subpopulations of T lymphocytes and Langerhans cells in oral lichen planus

This study has addressed the question of whether there is selective recruitment and distribution of intra-epithelial leucocytes in lesions of oral lichen planus (OLP). T-lymphocyte subsets were examined in the epithelium and peripheral blood of patients and controls using flow cytometry and double immunofluorescence, and the relationship between keratinocyte intercellular adhesion molecule-1 (ICAM-1) expression with T-lymphocyte and Langerhans cell (LC) distribution was examined. The circulating 'memory'subset (CD45RO⁺) of T-helper cells (CD4⁺) was increased from 49.1% in controls to 65.7% in patients ( P = 0.005), while the 'naive'subset (CD45RA⁺), which was absent from control epithelium, comprised 24% of helper cells in OLP ( P =0.037) and all T-cell and LC counts were significantly raised in ICAM-1-expressing areas of epithelium. These data demonstrate changes in intra-epithelial T-lymphocyte and LC populations compared with normal oral mucosa and suggest there is selective recruitment in OLP. In addition, Keratinocyte ICAM-1 expression does appear to be associated with accumulation of infiltrating T lymphocytes and LC.     

Enumeration of T cell subsets with monoclonal antibodies in minor salivary glands of patients with rheumatoid arthritis

Abstract – Labial salivary glands of 51 patients with rheumatoid arthritis and those of 25 control patients were examined by the ANAE (acid α-naphthyl acetate esterase) technique to determine the percentages of B- and T-lymphocytes and mononuclear phagocytes (MPS cells). Using monoclonal antibodies (OKT3 for all T cells, OKT4 for helper/inducer T cells, OKT6 for thymocytes, and OKT8 for suppressor/cytotoxic T cells) T cell subsets were enumerated. B- lymphocytes predominated in both series of salivary glands, and the percentages of B and T cells were equal in both series. The absolute cell counts in the salivary glands of rheumatics were significantly higher (P<0.001) than in those of healthy controls. The number of OKT4⁺ cells was increased in rheumatics, leading to an elevated OKT4⁺/OKT8⁺ ratio when compared with that in controls (P<0.01). The results suggest that the basic phenomenon behind the B cell hyperactivity noticed in rheumatics might be due to increased activity of T helper cells rather than reduced number of T suppressor cells, which were shown to remain almost unaffected in the salivary glands of patients with rheumatoid arthritis.     

Epithelial dendritic cells in pathological human oral tissues

Epithelial dendritic cells (EDC) were examined in human oral tissues with non-specific keratosis, lichen planus and squamous cell carcinoma. Acetone-fixed frozen sections were stained using an indirect immunoperoxidase technique and monoclonal antibodies to the human CD1 thymocyte (OKT6) and HLA-DR antigens. Significantly more T6⁺ and DR⁺ EDC were present in lichen planus tissues than normal controls, tissues with non–specific keratosis and the epithelia overlying/adjacent to squamous cell carcinomas, the latter tissues having comparable numbers of both T6⁺ and DR⁺ EDC. By contrast, significantly fewer T6⁺ EDC and significantly more DR⁺ cells were present in the invasive epithelium of squamous cell carcinomas than the overlying/adjacent epithelium of carcinomas, the non-specific keratosis group and the normal tissues. 23–60% of pathological tissues had either focal or general DR⁺ reactivity in keratinocytes, but there was no correlation between the density of T6⁺ or DR⁺ EDC and the keratinocyte DR status of the tissues. The results suggest that immunological enhancement occurs in lichen planus and possibly immunological impairment may characterize invasive squamous cell carcinoma.     

In situ characterization of mononuclear cells in human chronic marginal periodontitis using monoclonal antibodies

Acetone fixed cryostat sections from 25 patients with adult chronic marginal periodontitis were characterized using an indirect immunofluorescence technique with monoclonal antibodies. The amount of B lymphocytes (Leu-12 positive) varied considerably between the specimens and were usually seen in largest numbers in the most apical parts of the cellular infiltrates beneath the pocket epithelium (PE). Varying amounts of T lymphocytes (OKT 3 positive) were demonstrated in all specimens. The amount of T helper cells (OKT 4a positive) exceeded that of T suppressor/cytotoxic cells (OKT 8 positive) in the cellular infiltrates beneath the PE (OKT 4a/ OKT 8 =1.13). There was a more even distribution of these cell types beneath the oral gingival epithelium (OGE). Langerhans cells were observed within and occasionally subjacent to the OGE. Scattered macrophages (Leu-M3 or OK Ia 1 positive) were observed in the inflammatory cell infiltrates and on the connective tissue papillae beneath the OGE. HLA-DR antigen reacting with OK Ia 1 was present on cells corresponding to OKT 6 positive cells in the OGE and subjacent to the OGE as well as in the inflammatory cell infiltrates beneath the PE and in the perivascular infiltrates. In some specimens HLA-DR antigen was also found to be associated with keratinocytes in the outer parts of the OGE. Occasional NK cells (Leu-7 positive) were localized inside and subjacent to the OGE. There was a considerable variation with respect to the number and distribution of the various mononuclear cells between specimens and from section to section from the same specimen.     

In situ characterization of mononuclear cells in human dental periapical inflammatory lesions using monoclonal antibodies

Mononuclear cells in cryostat sections of human dental periapical inflammatory lesions were studied with the aid of murine monoclonal antibodies and with indirect immunofluorescence microscopy. T lymphocytes (OKT3-positive cells) made up a major part of the cells in the infiltrates. They were found mainly in clusters, although single cells were also seen. T helper cells (OKT4) were more numerous than suppressor/cytotoxic T cells (OKT8-positive cells), with a ratio of approximately 2:1. Langerhans cells (OKT6-positive cells) were not demonstrated: only a few scattered HNK 1-positive cells, probably natural killer cells, were detected. A large number of OKM1- and OKIa 1-positive cells were detected in the infiltrates. Their size and number varied considerably in the different areas of the sections. These cells are probably macrophages. Sheets of small OKIa 1-positive cells were also demonstrated, indicating the presence of B lymphocytes or activated T lymphocytes. The results indicate that immune reactions may be of importance in the pathogenesis of periapical inflammatory lesions.     

Inflammatory cell infiltrate associated with primary and transplanted tumours in an inbred model of oral carcinogenesis

This study characterised the nature of the local cellular immune responses associated with an inbred animal model of oral carcinogenesis. Inbred F344 rats developed moderately- to well-differentiated primary oral squamous cell carcinomas (SCC) after treatment with the carcinogen 4-nitroquinoline N-oxide (4-NQO) in vivo for 5–6 months. The inflammatory cell infiltrate associated with the primary tumours was predominantly of the macrophage lineage (CD45⁺, Ia⁺) and contained smaller numbers of CD8⁺ cells (NK cells, cytotoxic/suppressor T cells), CD5⁺ cells (T cells) and CD25⁺ cells (activated cells; T and NK cells). Keratinocyte cell lines were established from three lingual and one palatal SCC. By contrast to normal keratinocytes, tumour-derived cell lines were immortal and independent of 3T3 fibroblast support. All of the tumour-derived cell lines were tumorigenic in athymic ( nu/nu ) mice and showed contrasting latent periods of tumour development and histological differentiation; normal keratinocyte grafts were non-tumorigenic in athymic mice. Three of four malignant cell lines formed well-differentiated tumours in syngeneic F344 rats; the tumours regressed after 10–14 days. Regressing grafts contained significantly larger numbers of NK cells (CD5^-, CD8⁺) in the inflammatory cell infiltrate compared with that associated with primary tumours (p<0.04). One malignant cell line and normal keratinocytes were non-tumorigenic in syngeneic hosts. The results demonstrate phenotypic variation in the cell-mediated immune responses associated with the actively growing primary SCC and the regressing tumours in syngeneic hosts and suggest that NK cells, possibly activated by local T cell responses, are important for tumour rejection in this model.     

Quantitative analysis of immunocompetent cells in human normal oral and uterine cervical mucosa, oral papillomas and leukoplakias

Using monoclonal antibodies reacting with T-cell subpopulations, Langerhans cells and macrophages, the number and distribution of cells of the immune system in normal oral and cervical mucosa was determined and statistically compared with that in oral papillomas and oral leukoplakias. Increased numbers of labelled cells were found in oral leukoplakias and particularly in oral papillomas. In the epithelium of all specimens, Langerhans cells and T-lymphocytes of the suppressor/cytotoxic phenotype as well as of the helper phenotype were seen. Suppressor/cytotoxic and helper T-lymphocytes were in equal numbers in the epithelium of oral papillomas, but were about 2:1 in all other lesions. In normal oral epithelium, macrophages were rare but were in greater numbers in leukoplakias and papillomas. In the connective tissue of all lesions, more labelled cells were present than in epithelium with T-lymphocytes predominant. Although Langerhans cells were rare in connective tissue, many were seen in oral papillomas.     

Mucosal cell-mediated immunological changes associated with experimental graft-versus-host disease

This study examined the histological changes and local cellular immune response induced within the lingual mucosa in an allogeneic F, hybrid rat model of graft-versus-host disease (GvHD) with a view to studying oral lymphocyte-epithelial cell reactions. Highest levels of disease, as reflected by both a GvHD index and the extent of the oral mucosal changes, were obtained using primed donor (Lewis rats) splenocytes and irradiated hosts (Lew/Da rats). The lingual mucosae of test animals were characterised by irregular epithelial keratosis, an absence of basal cell liquefaction and a diffuse inflammatory cell infiltrate, histological features consistent with an oral lichenoid tissue reaction. Immunohistochemical studies showed that mucosal involvement was characterised by infiltration of the lamina propria by NK cells (CD8⁺, CD5⁻), "activated" cells (CD25⁺) and T cells (CD5⁺) with selective migration of the latter, including a CD5⁺. CD8⁻ subset (helper/inducer T cell), into the epithelium. Epithelial expression of la was invariably associated with these inflammatory cell infiltrates and correlated with the GvHD index. These findings suggest the presence of local mucosal T cell activation in the absence of detectable epithelial cell damage, which may be equivalent to the early initiating events in the pathogenesis of oral lichen planus. However, whilst experimental graft-versus-host disease appears to be a useful model for studying lymphocyte-epithelial interactions, the induced oral mucosal changes are more consistent with a lichenoid reaction rather than lichen planus.     

口腔颌面部固定性药疹的免疫组织化学研究

本文报道了１８例口腔颌面部固定性药疹病例,并应用４种鼠抗入白细胞抗原的单克隆抗体,采用标记的链霉亲本素生物素技术对其中１０例者急性期皮损内浸润的Ｔ细胞及其亚群、Ｂ细胞进行标记。     

Differentiation of T lymphocyte subpopulations, macrophages, and HLA-DR-restricted cells of apical granulation tissue

Sixteen specimens excised from apical granulation tissue were examined for immunocompetent T lymphocytes, mononuclear phagocytes, and HLA-DR-determined cells. Light microscopic visualization of the cells was performed using monoclonal antibodies and the immunocytochemical alkaline phosphatase anti-alkaline phosphatase technique. Quantitative assessment of the biopsy specimens showed that in five diagnosed cysts, eight granulomas, and three specimens taken from scar tissue, macrophages represented the dominating inflammatory cell fraction, followed by T lymphocytes. In the latter group, helper/inducer T cells were most frequent. Cysts showed a statistically significant increase in cell populations of helper/inducer, suppressor/cytotoxic T cells, macrophages, and HLA-DR-coded cells. There were only a few activated T lymphocytes.