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.     

Identification of Langerhans cells in human gingival epithelium

The purpose of this study was to qualitatively compare three recent techniques of Langerhans cells detection in oral epithelium and to quantitatively compare Langerhans cells in clinically normal and clinically inflamed human gingival biopsies. Eleven subjects were selected who displayed chronic periodontitis and moderate gingival inflammation. A quadrant associated with clinically inflamed tissues was not treated, while the remaining teeth were scaled and root-planed. Two gingival biopsies were taken: clinically normal, treated tissue; and clinically inflamed, untreated tissue. Langerhans cells were stained using HLD-DR, S-100 and OKT6. They were quantitated using a standard grid for OKT6-stained sections only. Approximately 5 times as many Langerhans cells were identified in the biopsy specimens of clinically inflamed human gingiva as in clinically normal gingiva of the same patient. Of the methods studied, OKT6 was qualitatively determined to be the best for visualization of these cells. An immunologic role in the host response to chronic periodontal disease is postulated for Langerhans cells.     

Characterization of mononuclear cells in cyclosporin A induced gingival enlargement

Abstract – Gingival biopsies from five patients with cyclosporin A (CsA) induced enlargement of the gingiva contained prominent mononuclear infiltrates beneath the epithelial basement membrane. These infiltrates and surrounding tissues were studied using monoclonal antibodies directed against lymphocytes and their subgroups (OKT 3, OKT 4, OKT 8, Leu 3 and Leu 4), monocytes (OKM 1) and the framework structure of HLA-DR antigens (OKI al). As revealed by the reactivity of these antibodies the vast majority of the mononuclear cells in two individuals consisted of monocytes and in the others of T-Iymphocytes. Virtually no B-lymphocytes were observed. The ratio between T-cells bearing T 4 and T 8 antigens, in separate biopsies, varied between 6 and 0.3. Control biopsies were obtained from the gingiva of seven healthy individuals without gingivitis. These biopsies contained relatively few mononuclear cells all of which had T-lymphocytc phenotype.     

Characterization of mononuclear cells in cyclosporin A induced gingival enlargement

Gingival biopsies from five patients with cyclosporin A (CsA) induced enlargement of the gingiva contained prominent mononuclear infiltrates beneath the epithelial basement membrane. These infiltrates and surrounding tissues were studied using monoclonal antibodies directed against lymphocytes and their subgroups (OKT 3, OKT 4, OKT 8, Leu 3 and Leu 4), monocytes (OKM 1) and the framework structure of HLA-DR antigens (OKI al). As revealed by the reactivity of these antibodies the vast majority of the mononuclear cells in two individuals consisted of monocytes and in the others of T-lymphocytes. Virtually no B-lymphocytes were observed. The ratio between T-cells bearing T 4 and T 8 antigens, in separate biopsies, varied between 6 and 0.3. Control biopsies were obtained from the gingiva of seven healthy individuals without gingivitis. These biopsies contained relatively few mononuclear cells all of which had T-lymphocyte phenotype.     

Langerhans cells in human chronic gingivitis and phenytoin-induced gingival hyperplasia

Langerhans cell numbers in oral epithelium increase as dental plaque accumulates. The anti-convulsant drug phenytoin predisposes to gingival hyperplasia in certain patients who take this medication for epilepsy and who also have poor oral hygiene. In this study 7 patients with phenytoin-induced gingival hyperplasia were compared with 5 subjects with chronic marginal gingivitis. On initial examination and on completion of the hygiene phase of periodontal therapy (a period ranging from 3.0 to 4.25 months), clinical indices of plaque and gingivitis were recorded and biopsies were taken from the lower anterior labial gingiva. Frozen sections were stained by an immunoperoxidase technique using the monoclonal antibody OKT6, and the number of Langerhans cells in a defined cross-sectional area was counted. In phenytoin-induced gingival hyperplasia there was a marked increase in Langerhans cells (13.8 +/- 0.45) when compared with chronic gingivitis (7.7 +/- 0.31; p less than 0.05). Both groups showed marked reductions in their plaque and gingival indices and numbers of Langerhans cells once treatment had been completed. However, levels of Langerhans cells in the drug-induced hyperplasia remained significantly higher (3.5 +/- 0.26) than in chronic gingivitis (1.5 +/- 0.22; p less than 0.05).     

Reduced numbers of Langerhans cells and increased HLA-DR expression in keratinocytes in the oral gingival epithelium of HIV-infected patients with periodontitis

BACKGROUND: HIV-seropositive (HIV+) patients become increasingly susceptible to periodontal diseases as HIV infection proceeds. We have previously shown that HIV+ patients with chronic marginal periodontitis (CMP) have remarkably increased numbers of gingival plasma cells in the connective tissue underlying the oral gingival epithelium, but depressed specific serum IgG levels towards periodontopathogenic bacteria. Langerhans cells (LC) and keratinocytes (KC) are antigen-presenting cells that are important in promoting immune responses. METHOD: In this study we examined, by means of immunofluorescence, the distribution and numbers of LC and activated KC in biopsies taken from inflamed periodontal sites in HIV+ and HIV patients with CMP. RESULTS: In the pocket epithelium in both patient groups, basal layer KC expressed HLA-DR molecules. In the oral gingival epithelium of HIV+ patients, basal layer KC also expressed HLA-DR molecules and numbers of LC were decreased as compared with HIV persons. CONCLUSION: The findings suggest that the oral gingiva in HIV+ patients may be affected by inflammation.     

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.     

Distribution of Langerhans cells in clinically healthy human gingival epithelium with special emphasis on junctional epithelium

Abstract – Twenty-one biopsies of clinically healthy marginal gingiva from children, who performed conventional oral hygiene but received no additional professional prophylaxis, were studied in order to obtain information on distribution and density of Langerhans cells (LC) in the oral gingival epithelium (OGE), the sulcular epithelium (SE) and the junctional epithelium (JE). A simple freeze-separation technique was found to create acceptable histomorphology of JE in specimens obtained adherent to teeth, while partially and non-adherent ones were rejected. The majority of LC in OGE, were highly dendritic and stained intensively with OKT6 monoclonal antibodies. The distribution was network-like with a density of 21.0± 3.2 LC/0.1 mm² crosssectional epithelial area. A similar although less dense distribution was found in SE (8.6 ± 3.0 LG/0.1 mm²). These observations, confirm previous findings. In JE 2 groups of LC were identified: 1) Weakly stained LC with very few and short dendrites distributed in a scattered way (2.8± 1.4 LC/0.1 mm²) in the apical three-fourths of JE in most specimens. Present evidence suggests that these cells might be immature cells of Langerhans lineage. 2) Clusters of LC (9.4 ± 2.9 LC/0.1 mm²) with dendrites of moderate lengths and numbers and a varied fluorescence intensity; they were found in a few specimens in the coronal one-fourth of JE and at the border zone to SE. Such clusters might represent genuine variation in the distribution of LC or reactions to initial/ early plaque formation.     

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.     

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.     

Endothelial cell leukocyte adhesion molecule-1 (ELAM-1) and intercellular adhesion molecule-1 (ICAM-1) expression in gingival tissue during health and experimentally-induced gingivitis

The changes in vascular adhesion molecule expression and numbers of infiltrating leukocytes during a 21-day experimental gingivitis episode were investigated immunohistochemically. Monoclonal antibodies to ELAM-1 (1.2B6), ICAM-1 (6.5B5), CD3 (OKT3-pan-T cell) and neutrophils (PMN-elastase) were used to identify positive vessels and leukocytes within gingival biopsies taken on d 0, 7, 14 and 21. Vascular endothelium expressed ELAM-1 and ICAM-1 both in clinically 'healthy' tissue (d 0) and in experimentally inflamed tissue (d 7 to 21). Positive vessels were found mainly in the connective tissue subjacent to the junctional epithelium where the highest numbers of T cells and neutrophils were also seen. Although T cells were found in all tissue areas studied, neutrophils were largely concentrated in the junctional epithelium and the subjacent connective tissue but were absent from the oral epithelial region. As the experimental gingivitis developed, the number of T cells or neutrophils in the different tissue regions did not change significantly although the most intense vascular ICAM-1 and ELAM-1 staining redistributed to the CT adjacent to the junctional epithelium. A prominent feature was the intense ICAM-1 positive staining of the junctional epithelium and its absence in the closely adjacent oral epithelium, in both clinically 'healthy' and inflamed tissue. The gradient of ICAM-1 in junctional epithelium, with the strongest staining on the crevicular aspect plus the vascular expression of ELAM-1 and ICAM-1 in both clinically 'healthy' and inflamed tissue may be crucial processes which direct leukocyte migration towards the gingival crevice.     

Host response to titanium dental implant placement evaluated in a human oral model

BACKGROUND: Recent reports have questioned if metal sensitivity may arise from exposure to titanium. The objective of this study was to histologically evaluate non-perforated mucosa covering submerged maxillary titanium implants with regard to induced tissue reactions. METHODS: Thirteen patients, 21 to 69 years of age, without previous implants were included. After initial examination, the bone crest areas destined for dental implant placement were exposed, and threaded external hex dental implants were inserted. Prior to wound closure, a full mucosal tissue slice was biopsied from the edge of the mucoperiosteal flap (baseline). The patients were monitored monthly for 6 months. At the abutment connection, biopsies were taken by a 6-mm punch, altogether yielding 26 specimens. Tissue reactions were analyzed by coded histometric analysis at four defined areas at increasing distance from the oral epithelium, including ratios of inflammatory cells (IC)/epithelial cells, IC/fibroblasts, and number of dense particles. RESULTS: The stained sections portrayed gingival tissue with intact oral epithelium and connective tissue with variable accumulation of IC. Experimental biopsies demonstrated mineralized areas and dense particles of different sizes. Analysis of variance revealed a higher IC/fibroblast ratio for level 3 at baseline compared to level 3 at 6 months (P<0.01). Furthermore, a significant decrease in IC/fibroblast ratio was observed between levels 2 and 3 and 2 and 4 at 6 months (P<0.001). The connective tissue level facing the cover screw contained the highest number of dense particles (P<0.01). CONCLUSIONS: Tissue sensitivity reactions to titanium implants were not disclosed. All 6-month biopsies contained dense particles that were most likely metals.     

Macrophage subpopulations in gingival overgrowth induced by nifedipine and immunosuppressive medication

BACKGROUND: The immunomodulating effects of both immunosuppressive and nifedipine medication have been associated with drug-induced gingival overgrowth. The aim of the study reported here was to evaluate the presence of macrophage subpopulations in normal human gingiva and in gingival overgrowth induced by nifedipine and immunosuppressive medication. METHODS: Gingival samples were taken from 11 nifedipine-medicated cardiac outpatients (nifedipine group), 11 triple-medicated organ-transplant recipients also taking nifedipine (immunosuppression plus nifedipine group), 12 triple-medicated organ-transplant recipients (immunosuppression group), and 20 generally healthy individuals (control group). Cryostat sections were stained with mAbs for inflammatory 27E10, reparative RM3/1, and resident 25F9 macrophages using an avidin-biotin enzyme complex method. Total numbers of mAb-labeled cells were determined in connective tissue beneath sulcular epithelium, connective tissue beneath oral epithelium, and middle connective tissue. Expression of 27E10 was determined in keratinocytes in the oral epithelium. Statistics analyses were undertaken using the chi-square test, the Mann-Whitney U test, the independent samples t test, analysis of variance, and analysis of covariance. RESULTS: Greater numbers of inflammatory 27E10-positive macrophages were found in all 3 medicated groups and counting zones than in the control group except in connective tissue beneath sulcular epithelium in the immunosuppression group. The incidence of specimens expressing 27E10 antigen throughout the oral epithelium was significantly higher in the immunosuppression group (8 of 12) than in the control group (4 of 20) and the nifedipine group (2 of 11). Numbers of reparative RM3/1-positive macrophages were significantly greater in the immunosuppression group in connective tissue beneath oral epithelium than in the control group. The effect was markedly associated with degree of inflammation. Numbers of resident 25F9-positive macrophages were lower in connective tissue beneath sulcular epithelium in the immunosuppression group, and higher in middle connective tissue in the nifedipine group than in the control group. CONCLUSION: Our results show that the nature of drug-induced gingival overgrowth differs somewhat between immunosuppressive and nifedipine medications.     

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.     

Human gingival Langerhans cells in health and disease

Epithelial Langerhans cells in samples of healthy and diseased gingival tissue were studied using ATPase histochemistry and the monoclonal antibodies OKT6 and anti HLA-DR. In healthy gingiva Langerhans cells were seen in both oral and sulcular epithelium; they were generally positioned in the basal layers. No Langerhans cells were seen in junctional epithelium. In diseased tissue there was a large increase in the number of Langerhans cells in both oral and sulcular epithelium with many more being situated in the stratum spinosum. There was an increase in the expression of the Class 2 antigen, HLA-DR, and morphological polarization occurred with dendrites preferentially orientated towards the surface. No Langerhans cells were seen in the pocket lining epithelium of periodontally diseased gingiva.     

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.     

Cytolethal distending toxin damages the oral epithelium of gingival explants

The cytolethal distending toxin (Cdt), expressed by the periodontal pathogen Aggregatibacter actinomycetemcomitans, inhibits the proliferation of cultured epithelial cells by arresting the cell cycle. The gingival epithelium is an early line of defense against microbial assault. When damaged, bacteria collectively gain entry into underlying connective tissue where microbial products can affect infiltrating inflammatory cells, leading to the destruction of the attachment apparatus. Histological evaluation of rat and healthy human gingival tissue exposed ex vivo to the Cdt for 36 and 18 hours, respectively, revealed extensive detachment of the keratinized outer layer and distention of spinous and basal cells in the oral epithelium. Treated human tissue also exhibited disruption of rete pegs and dissolution of cell junctions. Cells in the connective tissue appeared unaffected. Primary gingival epithelial cells, but not gingival fibroblasts, isolated from the same healthy human tissue were cell-cycle-arrested when treated with the toxin. These findings provide new evidence that the Cdt severely damages the oral epithelium, ex vivo, by specifically targeting epithelial cells, in situ. The Cdt shows preferential targeting of the epithelium as opposed to connective tissue in animal and human gingival explant models. Abbreviations: cytolethal distending toxin (Cdt), connective tissue (CT), 4',6-diamidino-2-phenylindole (DAPI), human gingival epithelial cells (HGEC), human gingival explants (HGX), human gingival fibroblasts (HGF), junctional epithelium (JE), oral epithelium (OE), rete pegs (RP), sulcular epithelium (SE).     

Langerhans cells in the gingival epithelium

Langerhans cells (LCs) were specifically demonstrated by monoclonal antibody OKT 6. In healthy gingival tissue LCs were mainly seen in stratum spinosum of the surface epithelium. They had small nuclei and with long cell processes. The LCs of oral epithelium in marginal gingivitis and adult periodontitis tissue were more in numbers than in healthy tissue. In juvenile periodontitis tissue LCs numbers seemed to be increased obviously in comparison to the healthy tissue. The LCs were round and often located in both deep spinous and basal layers. These results demonstrate that LCs play an important role in the local immune response of the periodontal tissues.     

Immunohistochemical identification of antigen presenting cells in rat salivary glands

Mucosal dendritic cells affect immune responses through secretion of cytokines and exposure of na?ve B- and T-lymphocytes to foreign matter as antigen presenting cells (APCs). APC in oral tissues may play a role in the development of local and secretory immune responses [Crit. Rev. Oral Biol. Med. 7 (1996) 36]. Previous studies have shown that APC are present in the interstitial tissues of rat salivary glands [Arch. Oral Biol. 40 (1995) 1015]. This study sought to further define the distribution of APC in salivary glands. The major glands and ducts of male Sprague-Dawley and Wistar rats were fixed with 4% paraformaldehyde and prepared for immunofluorescence and pre- and post-embedding immunoelectron microscopy. Monoclonal antibodies to the dendritic cell marker Ia antigen (OX-6 antibody), monocyte lineage cytoplasmic antigen (ED-1), and resident tissue macrophage antigen (ED-2) were visualized with FITC-conjugated secondary antibodies for light microscopy and HRP- and gold-labelled secondary antibodies for electron microscopy. Light microscopy revealed numerous OX-6-positive cells with branching processes in the epithelium of striated and excretory ducts of both rat strains, as well as in the connective tissue stroma. ED-1-positive cells had a similar distribution but exhibited a more compact shape with fewer processes. ED-2-positive cells were found only in the connective tissue. Acinar and duct epithelial cells were unreactive. Electron microscopy confirmed that both OX-6-positive and ED-1-positive, non-epithelial cells were present within the duct epithelium. The presence of APC in the duct epithelium suggests that these ducts may be exposed to antigens, possibly by retrograde access from the oral cavity, and that APC located in the salivary gland epithelium may participate in local immune responses.     

Immunohistochemical analysis of lymphocyte subpopulations in cyclosporin A-induced gingival overgrowth

BACKGROUND: Cyclosporin A (CsA) is an immunosuppressive agent that is known to induce gingival overgrowth (GO). Pharmacological, genetic, immunologic, and inflammatory factors seem to be involved in the complex pathogenesis of drug-induced GO. Lymphocyte subpopulations in human gingival connective tissue have been implicated in the pathogenesis of inflammatory periodontal diseases. One purpose of this study was to quantify CD4, CD8-, CD57-, and epithelial membrane antigen (EMA)-positive cells in the gingiva of renal transplant recipients treated with CsA, and compare them to findings in healthy controls. A second aim was to correlate cell numbers with clinical findings. METHODS: The study included 19 kidney recipients who were taking CsA and had significant GO (CsAGO+), 13 recipients who were taking CsA but showed no GO (CsAGO-), and 14 systemically healthy individuals with gingivitis (C). Sections from gingival biopsies were incubated with monoclonal antibodies for CD4, CD8, EMA, and CD57, and then analyzed using the avidin-biotin complex method. In each specimen, the mononuclear cell types were quantified and their distribution was evaluated in 3 separate tissue zones: S = subepithelial connective tissue beneath the sulcular epithelium; O = subepithelial connective tissue beneath the oral epithelium; and M = middle connective tissue. RESULTS: There were no significant differences among the groups with respect to the numbers of CD4+ and CD8+ cells in each of the 3 zones (P >0.05). In zone S, the CsAGO+ group had significantly more EMA-positive cells than either the C or CsAGO- groups (P <0.05). There were significant differences among the groups regarding numbers of CD57+ (natural killer) cells in zone M, with the lowest cell numbers in the CsAGO+ patients (P<0.05). CONCLUSIONS: The results showed that low numbers of natural killer cells are important in the expression of plaque-induced inflammatory changes in CsA-associated GO. It appears that these cells may influence the drug's ability to induce proliferative activity.