Noriyasu Murase

Heterogeneity of keratin distribution in the oral mucosa and skin of mammals as determined using monoclonal antibodies

SummaryThe immunohistochemical localization of keratins in the oral epithelia of several mammals was investigated using the monoclonal antibodies to keratins, PKK1 (41–56 kilodaltons) and KL1 (55–57 kilodaltons). The staining patterns obtained in different locations of the oral mucosa and of the skin epidermis were compared. In the papillae on the dorsal surface of the tongue, some areas exhibited marked PKK1 staining, while other area were PKK1 negative. In general, rodent oral epithelia were negative for PKK1 in the basal layer, while comparatively strong PKK1 staining was observed in cells of the upper spinous layer. In the epidermis, positive PKK1 reactions were confined to the basal layer, while KL1 staining was occasionally seen in the basal layer of oral epithelia. In cats, dogs, and monkeys, different PKK1 and KL1 binding patterns were observed in oral epithelia. Also, the distribution in oral epithelia differed from that seen in the epidermis of these animals. In the epidermis, the distribution of PKK1 and KL1 was regular, with PKK1 usually being confined to the basal layer, while KL1 binding was found in the spinous and granular cell layers, and was dependent on the degree of keratinization. In the animals studies, keratin expression as detected by PKK1 and KL1-was different in the skin epidermis and oral epithelia, and the localization of these keratins differed in the various types of oral mucosa.

Comparison of lectin binding patterns in salivary glands of mice and rats with special reference to different fixatives used

Lectin binding patterns in the salivary glands of mice and rats were compared among specimens treated with 9 kinds of fixatives and then individually subjected to 10 kinds of lectin staining. Formalin-fixed sections showed positive lectin binding with fine granular materials in the GCT cells, and alcohol- or acetone-fixed sections revealed vacuolated patterns of irregular lectin binding with insufficient morphologic detail. Bouins, Hellys, and Zambonis fixatives displayed an adequate distribution of lectin binding which corresponded to the histological aspects. Different lectins gave different characteristic binding patterns in SMGs of mice; i.e., PNA and SBA binding was positive in female GCT cells, but absent in the male. On the contrary these lectins gave positive binding in the male acinar cells but negative in the female. These contradictory results were obtained for PNA and SBA binding between the GCT cells and acinar compartments of the mouse SMGs. The GCT and acinar cells in the SMGs of mice and rats also gave contradictory results; i.e., mice GCT cells displayed positive Con A staining but negative PA/Con A staining, whereas mice acinar cells were stained weakly by the Con A and staining strongly by the PA/Con A methods. Rats GCT cells indicated negative Con A, and strong PA/Con A staining; whereas rats acinar cells gave a positive Con A and negative PA/Con A reaction.

Immunohistochemical localization of αl-antitrypsin and αl-antichymotrypsin in salivary pleomorphic adenomas

Immunohistochemical identification of αl-antitrypsin (αl-AT) and αl-antichymotrypsin (αl-ACh) in pleomorphic adenomas of salivary glands is reported in order to compare their distribution profiles with those of lysozyme and lactoferrin, already described elsewhere. Normal salivary glands indicated positive αl-AT staining in ductal segments and had no αl-ACh in any glandular cell. Pleomorphic adenomas displayed moderate positivity to αl-AT staining in duct-like, tubular and glandular epithelia which was particularly intense in luminal cells. The limited number of tumour cells which showed duct-like structures with a single cellular layer arrangement, displayed the highest staining to αl-ACh. Strongly αl-AT positive tumour cells located on the inner side of luminal cavities were also markedly positive to αl-ACh. Spindle shaped tumour cells existed outside tubular and ductal structures and were negative to αl-AT and αl-ACh. Distribution of αl-AT in salivary glands was similar to that of lysozyme as is usual in ductal segments or their transformed cells, and occurrence of αl-ACh localization rather resembled that of lactoferrin, with occurrence in acinar compartments and changed epithelia within acini. The biological role of a specific immunohistochemical distribution of αl-AT and αl-ACh in pleomorphic adenomas may be associated with a self regulating mechanism which inhibits degradation by tissue proteinases.

Immunohistochemical evaluation of factor VIII related antigen, filament proteins and lectin binding in haemangiomas.

Immunohistochemical identification of factor VIII related antigen (F VIII RAG) filament proteins (actin, myosin, filamin, vimentin and desmin) and lectin binding patterns of Con A, PNA, SBA, WGA, RCA-1, UEA-1 and DBA in the endothelial cells and the musclar layers of haemangiomas and normal blood vessels are reported, using paraffin sections with the HRP method. The endothelial cells of haemangiomas were usually strongly positive to F VIII RAG as were those from capillary vessels and other small vessels. Some of the endothelium from haemangiomas and angiokeratomas was negative for factor VIII. The vessel walls of hemangiomas showed staining slightly positive for microfilaments (actin, myosin, filamin). The smooth muscle layer in small vessels showed a more marked staining with actin. Vimentin and desmin reactions in the vessel walls of, haemangioma and in normal vessels were slight or moderate. UEA-1 lectin binding was constantly positive in endothelial cells from hemangiomas and in blood vessels. SBA and WGA binding appeared in the border layer of endothelium in haemangiomas and normal vessels.

Differential distribution of immunohistochemically detected keratin proteins in mammalian oral epithelia.

Keratin proteins were immunohistochemically demonstrated in different parts of the oral epithelium. Keratin staining in the squamous-cell epithelium was restricted to the spinous and granular cell layers, with a comparatively low reaction in the basal layer cells and none in the superficial cornified layer. In comparing the keratin staining levels, those in the buccal and sublingual epithelia were rather higher than those in the hard palatal epithelia. Staining intensities for keratin proteins were not the same in either different locations of the oral epithelium or in the same location in different animals.

Immunohistochemical demonstration of epidermal growth factor and nerve growth factor in experimental carcinogenesis in the mouse submandibular gland

SummaryImmunohistochemical demonstration of epidermal growth factor (EGF) and nerve growth factor (NGF) was made during chemical carcinogenesis in the mouse submandibular gland. The granular convoluted tubule cells in the normal male submandibular gland contained larger amounts of EGF and NGF than in the female. The initial phase and early stages in chemical carcinogenesis showed degranulation of the granular convoluted tubule cells with a marked decrease in EGF and NGF. Premalignant lesions such as duct-like structures and multicystic lesions showed variable staining for EGF and were usually negative for NGF. Material secreted into the luminal spaces revealed increased staining for EGF and NGF. Scattered tumor cells of the poorly differentiated squamous-cell carcinoma type and desquamated tumor cells contained abundant EGF, but no NGF. No positive reaction for EGF or NGF was found in the induced squamous-cell carcinoma cells.

Immunohistochemical demonstration of lysozyme and lactoferrin in salivary pleomorphic adenomas

SummaryImmunohistochemical identification of lysozyme and lactoferrin was made in salivary pleomorphic adenomas (147 cases) and the staining patterns were evaluated with respect to the histological features and histogenesis. In normal salivary glands, the intercalated duct cells gave positive staining for lysozyme in major glands, and serous acinar cells, demilune cells, and interlobular duct cells were positive in minor glands. Lactoferrin staining was irregularly positive in serous cells and ductal epithelium. In pleomorphic adenomas, the reaction for lysozyme was positive in 14% (21/147) of the cases, and was confined to luminal cells of tubulo-ductal structures. Lactoferrin in pleomorphic adenomas was distributed in luminal tumor cells (51%; 75/147), in outer tumor cells (3%; 4/147), and in both luminal and outer tumor cells (5%; 7/147) in tubulo-ductal structures; it was also detected in plasmacytoid myoepithelial cells (5%, 8/147). However, modified myoepithelial cells and other types of neoplastic myoepithelial participants were negative for lactoferrin staining. The occurrence of both lysozyme and lactoferrin in salivary pleomorphic adenomas suggests their participation in the local defense mechanism in the tumor.

Keratin distribution in precancerous stages of experimental carcinogenesis in mouse submandibular glands

SummaryThe immunohistochemical distribution of keratin is reported in experimental carcinogenesis in the mouse submandibular gland (SMG). The initial changes included degranulation of granular convoluted tubule (GCT) cells and the appearance of keratin in the degranulated cells. There was a gradual increase in the area showing keratin staining in the altered tubule cells. Duct-like and cystic structures exhibited an intense keratin staining of their lining epithelium. The squamous cell carcinomas induced varying degrees of keratinization and positive immunohistochemical keratin staining. The latter technique provided a useful marker for distinguishing tumor cells of segmental duct origin in the salivary gland.

Keratin proteins identified in epithelial fragments of salivary mucoceles

Immunohistochemically detected keratin proteins in mucoceles of oral mucosa were used to served as markers to identify epithelial-derived cells of minor salivary glands. Normal ducts of minor salivary glands showed an intense keratin staining. Epithelial lining and or epithelial fragments in cystic walls of mucoceles displayed a strong reaction to keratin proteins too, whereas granulation tissue or connective tissue fibers of the walls were not seen. Foamy macrophages present in cystic cavities signify higher staining with the use of keratin proteins. Mucoceles in oral minor salivary glands are probably caused by ductal obstructions leading to continuous mucous secretion. Keratin proteins were used as an epithelial marker of ductal segments.

Carcinoembryonic antigen in mucoceles of oral mucosa

Immunohistochemical detection of carcinoembryonic antigen (CEA) was reported in mucocele and associated minor salivary glands of oral mucosa. Higher levels of staining for CEA occurred in salivary glands with attached mucoceles, in which acinar cells and ductal segments in the gland displayed concentrations of CEA higher than those in normal minor salivary glands. Floating cells also gave positive CEA staining, whereas epithelial fragments and connective tissue in the mucocele wall were lacking in CEA. It was suggested that mucoceles in oral mucosa accumulated CEA in associated minor salivary glands.