Differentiation of Langerhans Cells from Interdigitating Cells Using CD1a and S-100 Protein Antibodies

The present study shows that Langerhans cells can be differentiated from Interdigitating cells at the light microscopic level. Superficial lymph nodes and skin taken from necropsies and the lymph nodes of dermatopathic lymphadenopathy (DPL) were used for this experiment. Sections of lymph node and skin were embedded using the acetone, methyl benzoate and xylene (AMeX) method and dendritic cells were immunostained with anti S-100 protein antibody (S-100, and OKT-6 (CD1a) using the restaining method. Langerhans cells in the skin were positive for both CD1a and S-100. Dendritic cells positive for both CD1a and S-100, and dendritic cells positive for S-100, but not for CD1a were observed in superficial lymph nodes. In normal superficial lymph nodes, there were more interdigitating cells than Langerhans cells. The majority of the dendritic cells in the DPL were Langerhans cells. We conclude that the S-100 and CD1a positive cells are Langerhans cells, and the S-100 positive-CD1a negative cells are interdigitating cells.     

Immunocytochemical detection of dendritic cell by S-100 protein in the chicken.

Positivity for S-100 protein in paraffin embedded chicken lymphoid tissue was found by using a polyclonal antibody against whole bovine S-100 protein. The S-100 protein-containing cells were observed in the locations which have been reported to contain avian dendritic cells such as the medulla of the bursal follicles, and the germinal centers and T-dependent areas in the spleen, Peyer's patches, caecal tonsil and Harderian gland. Positive cells were also found in the location where ellipsoid associated cell have been described, and between epithelial cells covering the Peyer's patches and the caecal tonsil, as well as between the cells lining the ducts within the Harderian gland. Macrophages were devoid of immunostaining. Our results confirm the location described elsewhere for chicken dendritic cells and indicate that S-100 protein can be considered as a cell marker for the identification of the chicken dendritic cell. Intraepithelial positive cells may be interdigitating dendritic cells in an unusual location (their function being the transport of the antigen from the epithelium to the diffuse lymphoid tissue), or cytotoxic T-lymphocytes which, in mammals, are immunoreactive for S-100 protein.     

Differentiation of Langerhans Cells from Interdigitating Cells Using CD1a and S-100 Protein Antibodies

The present study shows that Langerhans cells can be differentiated from Interdigitating cells at the light microscopic level. Superficial lymph nodes and skin taken from necropsies and the lymph nodes of dermatopathic lymphadenopathy (DPL) were used for this experiment. Sections of lymph node and skin were embedded using the acetone, methyl benzoate and xylene (AMeX) method and dendritic cells were immunostained with anti S-100 protein antibody (S-100, and OKT-6 (CD1a) using the restaining method. Langerhans cells in the skin were positive for both CD1a and S-100. Dendritic cells positive for both CD1a and S-100, and dendritic cells positive for S-100, but not for CD1a were observed in superficial lymph nodes. In normal superficial lymph nodes, there were more interdigitating cells than Langerhans cells. The majority of the dendritic cells in the DPL were Langerhans cells. We conclude that the S-100 and CD1a positive cells are Langerhans cells, and the S-100 positive-CD1a negative cells are interdigitating cells.     

Immunohistochemical demonstration of S-100 protein in epithelial cells of bovine oviduct

The present study deals with an immunohistochemical localization of S-100 protein in the bovine oviduct. The epithelium of the infundibulum, ampulla and isthmus showed a positive staining for S-100 protein. The immunoreactivity for S-100 was observed both in the ciliated and nonciliated (secretory) cells of the oviductal epithelium at any stages of the estrous cycle. The immunoreactivity was also found in nervous elements and endothelial cells of blood vessels. No cell outside these cells showed any immunoreactivity for S-100. Although the functional significance of S-100 protein in the oviductal epithelium remains to be elucidated, the present results introduce new perspectives into the investigation of function and localization of S-100 protein.     

Intermediate filament expression in non-neoplastic pituitary cells.

Fifty-one non-neoplastic human pituitary glands, including examples with Crooke's hyalinization or amyloidosis, were examined by an immunoperoxidase method using antibodies to keratin, vimentin, neurofilaments (NFs), glial fibrillary acidic protein (GFAP), desmin, actin, S-100 protein and a variety of pituitary hormones. It was confirmed that most of the epithelial cells in the pituitary gland express keratin immunoreactivity. These cells included endocrine cells in the anterior lobe, endocrine cells and squamous metaplastic cells in the pars tuberalis, columnar and ciliated epithelia forming follicular structures and salivary-type epithelium in the pars intermedia, and anterior lobe cells infiltrating the posterior lobe. This study also demonstrated that keratin and NFs may be co-expressed in endocrine cells in the pituitary anterior lobe, that keratin, vimentin and GFAP may be co-expressed in the epithelial cells forming cyst-like follicle in the pars intermedia, and that vimentin and GFAP may be co-expressed in folliculo-stellate cells and pituicytes. In addition, the GFAP and S-100 protein-negative high columnar epithelium in the pars intermedia tended to be positive for adrenocorticotropic hormone and melanocyte stimulating hormone, while the low columnar epithelium with the co-expression of GFAP and S-100 protein was negative for pituitary hormones.     

S-100 protein subunits in bovine oviduct epithelium:In situ distribution and changes during primary cell culture

Summary Cultures of bovine oviduct epithelial cells are widely used in co-culture systems to improve the results ofin vitro fertilization. The aim of the present study was to evaluate the suitability of S-100 protein as a differentiation marker for bovine oviduct epithelial cellsin vitro. The distribution of S-100α and S-100β was examined immunohistochemically in bovine oviduct epitheliumin situ and in primary cell cultures derived from it. Three segments of the Fallopian tube (isthmus, ampulla and fimbriae) were compared and analysed during different stages of the oestrus cycle (luteal phase and follicular phase). Ciliated and non-ciliated cells of the epithelium reacted with anti-S-100α, S-100a(αβ) and S-100β antibodies, except for isthmic non-ciliated cells, which did not bind anti-S-100β or anti-S-100a(αβ). In addition, basal cells never showed immunoreactivity for S-100. In confluent monolayers of cultured oviduct epithelial cells, disappearance of reactivity for S-100 paralleled morphological signs of dedifferentiation (loss of cilia, cytoplasmic vacuolization). Free-floating oviduct epithelial cells, in contrast, retained morphological differentiation and still expressed S-100 antigen even after seven daysin vitro. The immunohistochemical findings were confirmed by polyacrylamide gel electrophoresis and Western blotting. The results indicate that the presence of S-100 is closely connected to morphological differentiation and to the specific functional condition of bovine oviduct epithelial cells.     

Langerhans cells in odontogenic tumours and cysts as detected by S-100 protein immunohistochemistry

Immunohistochemical demonstration of S-100 protein in Langerhans cells (LCs) was made in odontogenic epithelial tumours (71 cases), radicular cysts (40 cases), follicular cysts (28 cases), odontogenic keratocysts (11 cases), primordial cysts (7 cases) and fissual cysts (6 cases). With the use of polyclonal antiserum against S-100 protein, positive LCs, dendrical or irregular in shape were found in tumour or cystic epithelia, and sometimes in stromal connective tissue. Incidence of positive S-100 staining LCs was 11 cases out of 61 ameloblastomas, 22 cases out of 40 radicular cysts, 3 cases of 28 follicular cysts, and other lesions in both odontogenic tumours and cystic diseases lacked LCs. The cases with S-100 protein positive LCs were usually accompanied with a high degree of inflammatory infiltration in their lesions; on the contrary, the negative cases also generally lacked inflammatory responses.     

Role of bombesin receptor activated protein in the antigen presentation by human bronchial epithelial cells

Bombesin receptor activated protein (BRAP) was identified in a bacterial two‐hybrid screen for proteins interacting with bombesin receptor subtype‐3 (BRS‐3). We found that BRAP is widely expressed in the airway epithelium of human lungs and may play a role during the stress response of lung epithelium. In this work, we explored the potential roles of BRAP in the antigen presenting function of human bronchial epithelial cells (HBECs). Overexpression of a BRAP recombinant protein in a human bronchial epithelial cell line resulted in a reduction of FITC‐OVA uptake by HBECs, which indicated that the antigen uptake ability is inhibited. The analysis of the protein expression of surface molecules including B7 homologs and the major histocompactability complex (MHC) class II molecules showed that the expression levels of HLA‐DR and B7DC increased while the levels of B7‐H1 and B7.2 decreased. Since those surface molecules are all related to antigen presenting process, the altered expression pattern of those molecules provides further evidence showing that BRAP overexpression leads to a change in antigen presenting function of HBECs. Moreover, overexpression of BRAP in HBECs caused a decrease of co‐cultured lymphocytes proliferation and a changed pattern of cytokines produced by lymphocytes in the presence of FITC‐OVA, which indicated that changes in the maturation pattern and functions of co‐cultured lymphocytes were induced by BRAP overexpression. Overall, our results suggested that overexpression of BRAP may play a role during the antigen presenting process of bronchial epithelium by inhibiting the antigen uptake ability of bronchial epithelial cells. J. Cell. Biochem. 114: 238–244, 2012. © 2012 Wiley Periodicals, Inc.     

Calcifying epithelial odontogenic tumor: an immunohistochemical case study

Calcifying epithelial odontogenic tumor (CEOT) is a rare benign odontogenic tumor. A case of CEOT in a 25-year-old female is presented here. Histologically, the case showed sheets of polyhedral epithelial cells with deep eosinophilic cytoplasm and prominent nuclei. Nuclear pleomorphism and hyperchromatism were evident. Globules of amyloid-like material among the tumor cells were prominent. Also found was a small area demonstrating a cribriform pattern. Immunohistochemical studies with antibodies against basement membrane proteins (laminins 1 and 5, collagen type IV and fibronectin), pan-cytokeratins AE1/AE3, vimentin, S-100 protein and CD 1a were performed. Tumor cells expressed laminins 1 and 5, fibronectin, cytokeratins and vimentin. The amyloid-like material was not reactive to all antibodies examined. A number of dendritic cells among sheets of tumor cells were revealed with strong staining for S-100 protein and CD 1a. These dendritic cells are likely to be Langerhans cells. Hence, immunohistochemistry is a useful method to study the variant of CEOT.     

Dendritic cells in bronchial adenocarcinoma

We studied 28 bronchial adenocarcinomas (including 11 bronchiolo-alveolar carcinomas) stained for S-100 protein by the PAP technique in order to visualize dendritic cells in the neoplastic area. Dendritic cells were found in all cases varying in number from single to multiple and from region to region. Bronchiolo-alveolar carcinomas contained a similar quantity of these cells. The most numerous dendritic cells up to several scores per microscopical low-power field almost in all fragments of the tumour were found in three adenocarcinomas with numerous foci of the squamous cell metaplasia. Normal and metaplastic squamous epithelium of the bronchial mucosa did not show the presence of dendritic cells.     

Significance of S-100 protein immunostaining in the immunohistological analysis of normal and neoplastic lymphoid tissues--an appraisal

S-100 protein is a heterogeneous fraction of dimeric polypeptides (alpha and beta subunits) that can exist in different combination forms within the various tissues. Concerning the S-100 protein immunodetection within lymphoid tissue, the heterogeneity of the S-100 antigen, the tissue quality (frozen or paraffin-embedded after treatment with different fixatives) and the treatment of the tissue with different immunostaining methods and antibodies of different nature, all make for inconsistent results obtained in the immunohistological studies reported in the literature. Most of the S-100-positive cells of the lymphoreticular system are dendritic cells involved in the immune response (interdigitating reticulum cells, Langerhans cells, and follicular dendritic reticulum cells), other S-100-positive cells belonging to the mononuclear/phagocytic system. S-100 protein immunostaining may be used as a helpful immunohistological diagnostic clue to certain malignancies of the immune system (follicular center cell lymphomas) on the basis of their specifically related dendritic cell microenvironment. In addition to monoclonal antibodies for the immunophenotypic characterization of dendritic cells and macrophages and to enzyme reactions, the combined use of anti-S-100 antibodies specific for each of the S-100 protein subunits, tested with sensitive procedures, would be a very useful tool in the attempt to classify the proliferative disorders of dendritic cells and macrophages.     

Warthin's tumor as a hamartomatous dysplastic lesion: a histochemical and immunohistochemical study.

The etiology of Warthin's tumor was sought by histochemical and immunohistochemical methods using 7 surgically extirpated samples and normal salivary glands as a control for the epithelial component. All the samples exhibited a variety of amyloid deposition in the interfollicular area of the lymphoid component. The interfollicular lymphoid cells were both T-cells and cells of B-cell lineage with an almost 1 to 2 population ratio. Most antigen-positive B-cells were plasma cells that exhibited polyclonality of immunoglobulin. B-cells were also present in the lymphoid mantles and a few were found in the germinal centres. The epithelial component exhibited mucinous and proteinaceous fluid in the lumen and varied immunohistological reactions; being particularly positive to carcinoembryonic antigen, S-100 protein, and B-cell antigen; quite similar to that of normal salivary duct cells. The results suggest that Warthin's tumor may not be a hamartomatous neoplasm at all but a hamartomatous dysplastic lesion.     

Aberrant expression of HLA-DR antigens of the MHC class II in bronchial epithelial cells in asthmatic patients

The study of cellular events in the bronchi of asthmatic patients shows significant epithelial destruction. The ciliated cells are more often destroyed than others in the respiratory epithelium. Using highly specific monoclonal antibodies, we detected that the epithelial cells were positive for HLA-DR antigen expression, whereas those from healthy subjects were negative. The aberrant expression could be explained by the presence of T lymphocytes in the asthmatic bronchial mucosa. The activated lymphocytes (CD4+) are known to release gamma interferon, which is a lymphokine that induces or enhances the expression of HLA-DR antigens. These cells, which are known to mediate cytotoxicity in an Ag-specific and Ia-restricted way, could take part in the shedding process of epithelial cells in asthma.     

Differential distribution of immunoreactive S-100 protein in mammalian testis

The present study deals with the immunohistochemical localization of S-100 protein in the testes of seven mammalian species including rat, cat, dog, pig, sheep, cattle and horse. Significant differences are demonstrated in the cellular distribution and intensity of immunoreaction for the protein. In bull, ram, boar and cat tests S-100 protein was localized in the cytoplasm and nuclei of Sertoli cells. A particularly intense staining was seen in the modified Sertoli cells of the terminal tubular segment. With the exception of the cat and horse S-100 protein immunoreactivity was additionally found in epithelial cells of the straight testicular tubules and in the epithelial cells of the rete testis. Endothelial cells of capillaries, veins and lymphatic vessels are regularly S-100 immunoreactive in ruminants. Leydig cells were found to be strongly positive for S-100 protein in the cat and rat testes and to a lower degree in pig and horse testes. Finally a distinct immunostaining of peritubular cells was restricted to the testis of dogs and rats. The remarkable species-specific variations of immunoreactivity for S-100 protein in different cell types of the testis support the hypothesis that S-100 protein is a multifunctional protein and may have a different function in testicular physiology.     

Immunohistochemical study of a case of malignant müllerian mixed tumor in comparison with the activity of normal uterine tissue

A case of malignant Müllerian mixed tumor of the uterus, exhibiting a histology of heterologous osteosarcomatous differentiation, is presented. Special emphasis is placed on the characteristic immunohistochemical reactivity of the tumor tissue in comparison with that of normal uterine tissue in the proliferative phase. Keratin, cytokeratin, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA), and human chorionic gonadotropin (HCG) were found only in the carcinomatous element. However, neuron specific enolase (NSE), S-100 protein (S 100) and vimentin were identified in almost all tumor tissue elements. Desmin and actin were not stained in any elements. Myoglobin was only detected weakly in the squamous carcinomatous element. Undifferentiated cell element, composed of small, round, spindle, or polygonal cells showed positive reactions to NSE and S 100, but not to any other antibodies. As compared to the reactivities of the normal proliferative endometrium, the glandular epithelial cells were positive with NSE, S 100, vimentin and CEA, but the stromal cells were only positive with vimentin. Such a multitudinous and concomitant expression of antigenicity to the different tumor elements indicates a close relationship to its mesodermal Müllerian origin, and NSE, S 100 and vimentin might be most adequate indicators of these types of tumors.     

Fine needle aspiration of Langerhans cell histiocytosis of the lymph nodes. A report of six cases

BACKGROUND: Fine needle aspiration (FNA) diagnosis of Langerhans cell histiocytosis (LCH) of the lymph nodes has been described rarely. CASES: LCH was confined to the lymph nodes in six children. The FNA smears showed high cellularity composed of many isolated Langerhans cells (LCs) with nuclear grooves and intranuclear inclusions. Also seen were numerous eosinophils, lymphocytes, giant cells, dendriticlike cells, macrophages and Charcot-Leyden crystals in a background of eosinophilic granules. Immunohistochemical study revealed a positive reaction with S-100 protein. CONCLUSION: The presence of LCs with nuclear grooves, eosinophils, giant cells and a positive reaction with S-100 protein aided the diagnosis of LCH of the lymph nodes. Charcot-Leyden crystals, intranuclear inclusions and dendriticlike cells were seen in this study. These findings have not been reported before in lymph node FNA smears of LCH.     

Mucoepidermoid carcinomas: immunohistochemical studies on keratin, S-100 protein, lactoferrin, lysozyme and amylase

Immunohistochemical expression of 8 cases of mucoepidermoid carcinomas (G-I, 3 cases; G-II, 2 cases; and G-III, 3 cases) revealed marked heterogeneity of the proteins examined. Immunohistochemically detectable keratins (TK, KL1, and PKK1) were distributed in epidermoid cells, but were absent in mucous secreting cells. Strongly positive deposits of keratin proteins were detected in squamoid tumor cells in the G-I tumors. The tumor cells displayed positive staining for S-100 alpha, but did not stain with polyclonal S-100 antiserum or with monoclonal S-100 beta. The cells showing highest reactivity for S-100 protein were scattered in neoplastic foci and were probably Langerhans cells. Lactoferrin and lysozyme reactions were generally negative in tumor foci; but a positive reaction for lactoferrin was found in luminal tumor cells although rarely, and lysozyme staining was occasionally noted in histiocytes in the stroma. Amylase activity was usually absent in the tumor cells, with the exception of one case in which it was confined to the tumor cells. Mucoepidermoid carcinomas of various grades indicated marked heterogeneity in terms of various immunohistochemically detectable proteins.     

Immunostaining of a cell type in the islets of Langerhans of the monkey Macaca irus by antibodies against S-100 protein

Summary S-100 protein-immunoreactive cells were demonstrated by immunocytochemical procedures in the pancreatic islets of Langerhans in the monkey Macaca irus. By use of antibodies against human S-100 protein or bovine S-100 protein, these cells were observed in all islets in the head and tail portions of the pancreas. Immunostained cells were usually located in the center of the islets or sometimes found in a more widely distributed form, but they were never arranged in a regular concentric fashion. The number of immunoreactive cells varied from one islet to another but it was relatively limited making up only 0.75%–6.3% of all insular cells. With the use of the double-immunoenzymatic procedure for demonstration of the four main endocrine cell types (insulin-, glucagon-, somatostatin-and pancreatic polypeptide producing elements), it was possible to establish that S-100 protein-immunoreactive cells represent a distinct cell type. Antibodies against S-100 protein-stained neuroinsular complexes. The present findings speak in favor of a new cell type to be added to the large variety of S-100 protein-immunoreactive cells outside the central nervous system.     

Nasal Epithelial Cells Can Act as a Physiological Surrogate for Paediatric Asthma Studies

Introduction

Differentiated paediatric epithelial cells can be used to study the role of epithelial cells in asthma. Nasal epithelial cells are easier to obtain and may act as a surrogate for bronchial epithelium in asthma studies. We assessed the suitability of nasal epithelium from asthmatic children to be a surrogate for bronchial epithelium using air-liquid interface cultures.

Methods

Paired nasal and bronchial epithelial cells from asthmatic children (n = 9) were differentiated for 28 days under unstimulated and IL-13-stimulated conditions. Morphological and physiological markers were analysed using immunocytochemistry, transepithelial-electrical-resistance, Quantitative Real-time-PCR, ELISA and multiplex cytokine/chemokine analysis.

Results

Physiologically, nasal epithelial cells from asthmatic children exhibit similar cytokine responses to stimulation with IL-13 compared with paired bronchial epithelial cells. Morphologically however, nasal epithelial cells differed significantly from bronchial epithelial cells from asthmatic patients under unstimulated and IL-13-stimulated conditions. Nasal epithelial cells exhibited lower proliferation/differentiation rates and lower percentages of goblet and ciliated cells when unstimulated, while exhibiting a diminished and varied response to IL-13.

Conclusions

We conclude that morphologically, nasal epithelial cells would not be a suitable surrogate due to a significantly lower rate of proliferation and differentiation of goblet and ciliated cells. Physiologically, nasal epithelial cells respond similarly to exogenous stimulation with IL-13 in cytokine production and could be used as a physiological surrogate in the event that bronchial epithelial cells are not available.     

Secretory differentiation and cell type identification of a human fetal bronchial epithelial cell line (HFBE)

A human fetal bronchial epithelial cell line (HFBE) grew in an undifferentiated pattern under conventional culture conditions. Despite a somewhat fibro-blastic shape the cells maintained immunoreactivity to cytokeratin, carcinoembryonic antigen and epithelial membrane antigen. When grown on a collagen gel in a growth-hormone-supplemented medium, their spindle shape became more conspicuous. With an additional supplement of vitamin A (6 μg/ml), most of the cells underwent differentiation by producing many bright inclusion bodies which proved to be strongly positive with periodic acid-Schiff and weakly positive with alcian blue staining. Electron microscopy revealed a well-developed rough endoplasmic reticulum, an enlarged Golgi apparatus and many highly electron-dense secretory granules resembling those of Clara cells. Biochemical analysis demonstrated that HFBE cells cultured on collagen gel with vitamin A secreted hyaluronic acid and neutral glycoproteins containing mainly N-linked glycoproteins whose glycans were of a complex type. A monoclonal antibody (SEC-41) generated against the neutral glyco-proteins detected a glycoprotein of approximately 52 kDa in the spent culture medium of differentiated HFBE cells. This antibody also reacted with the intracytoplasmic secretory granules in these cells. When tested on frozen sections of lung tissue, the immunohistochemical reactivity of the SEC-41 antibody was confined to Clara cells, some type II pneumocytes in the adult lung, and respiratory epithelial cells in the fetal lung. More-over, this antibody could detect secretory glycoprotein in broncho-alveolar lavages from two patients. This paper clearly demonstrates that cells derived from human fetal bronchial epithelium can be cultivated in an undifferentiated precursor state and, under appropriate culture conditions, can be stimulated to undergo differentiation into a Clara cell type.