Matsuji Hosaka

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.

Lectin binding patterns in salivary glands treated with amylase.

Lectin binding patterns of ConA (Glc, Man), PNA and SBA (Gal, GalNAc), RCA-I (Gal) DBA (GalNAc), WGA (GlcNAc), and UEA-I (Fuc) in the major salivary glands of mice, rats, hamsters, and guinea pigs were reported using paraffin sections subjected to alpha-amylase treatment at 1, 3, and 6 h digestions. Lectin staining following treatment with amylase was generally enhanced in acinar, duct, and GCT cells. However, increasingly different reactions were obtained depending upon the lectins used, the various salivary glands from different specimens treated, and the different properties of the serous, mucous, and sero-mucous cells in the histologic sections. The lectins that demonstrated rather markedly increased staining were ConA, PNA, SBA, WGA, and UEA-I, whereas RCA-I and DBA increased little in comparison, or actually decreased. It appears from these findings that complex carbohydrates within murine salivary glands contained large amount of glucose, mannose, galactose, and N-acetyl galactosamine residues. The basement membranes of glandular cells in salivary glands demonstrated markedly positive ConA staining following alpha-amylase digestion.

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.

Heterogeneous expression of S-100 protein subunits α and β in cystadenolymphomas of salivary glands

Summary Immunohistochemical expression of S-100 protein and its subunits α and β in a total of 27 cases of cystadenolymphomas of the parotid gland is described by means of polyclonal anti-S-100 protein antiserum and monoclonal antibodies against S-100 α and β. Normal salivary glands showed strongly positive staining for S-100α in acinic cells and a negative or slightly positive reaction in ductal cells; however, they were usually negative for S-100β staining. Neoplastic epithelial cells of cystadenolymphomas showed slightly positive staining for S-100α in their cytoplasm, and there was a strongly positive reaction in a limited number of tumour cells. The S-100β subunit was expressed as a fine granular deposition in the apical cytoplasm of tumour cells. S-100 protein staining with the polyclonal reagent was usually negative or slightly positive in the tumour epithelia, but a small number of strongly positive cells were scattered throughout the tumour epithelia, suggesting the presence of Langerhanss cells. Interdigitating cells in lymphoid tissue reacted positively to monoclonal S-100α as well as to the polyclonal antiserum.

Neuroblastoma of parotid gland: Report of a case and immunohistochemical characteristics

A case of a parotid mass in a 2-year-old boy, postoperatively diagnosed as neuroblastoma, a rare tumour not previously reported in the parotid gland is presented. The neoplasm developed within the parotid gland as a painless mass without regional lymphadenopathy. Histopathologically, the tumour showed primitive nerve cells-neuroblasts-with round or oval dark basophilic nuclei and scanty cytoplasm. The cells were arranged in circular rosettes around an eosinophilic mass consisting of very fine filaments originating in the tumour cells or papillary configuration and sometimes scattered in the poorly developed stroma. Immunohistochemical evaluation of the tumour showed a positive immunoreactivity for vimentin, alpha and beta subunits of S-100 protein, neurone-specific enolase (NSE), substance P, met-enkephalin and chromogranin but cytokeratins, desmin, actin, myosin, glial fibrillary acidic protein (GFAP) and calcitonin gene related peptide (CGRP) were negative. The histopathological and immunohistochemical findings conclude a diagnosis of neuroblastoma of the parotid gland.

Distribution difference of lectin binding in salivary gland treated with sialidase and trypsin

Histochemical evaluation of complex carbohydrates in the salivary glands from rodents following sialidase and trypsin treatment were reported by the use of lectins binding technique which specifically link to corresponding sugar residues in macromolecules. Lectin staining in salivary gland generally increased following 1 h sialidase treatment, particularly in guinea-pig specimens. Lectin staining followed by treatment of trypsin (30 min) showed in increase in staining which is characteristic of UEA-I lectin. In long duration treated sections, by either sialidase or trypsin, lectin staining usually decreased in salivary glands. In the present study, complex carbohydrates of salivary glands may be hydrolyzed and degraded by sialidase and trypsin treatments, and lectin binding affinity is then also enhanced due to exposed sugar residues in complex carbohydrates.

Branchial cleft cysts. Histologic and immunohistochemical aspects.

8 cases of branchial cleft cysts and 1 case of branchiogenic carcinoma were examined immunohistochemically for detectable keratin, immunoglobulins (IgG, IgA, IgM), carcinoembryonic antigen (CEA), factor VIII related antigen (Factor VIII RGA), and lysozyme. Lectin binding patterns were also determined. Histologically, cystic lining epithelia were classified into stratified squamous epithelium without keratinization, columnar epithelium with or without cilia, or a mixture of both. Almost all of the cases indicated accompanying lymphoid structures with germinal centers. Keratin expression in epithelial cells was slightly positive, and lectin binding affinities in them were similar to those of oral squamous epithelium. CEA was found on the surface border of columnar epithelial cells, but cystic epithelia in most of the cases were devoid of lysozyme. Endothelial cells of capillary vessels showed positive binding by UEA-1 lectin and the presence of factor VIII RAG. In the lymphoid structures, there were scattered strongly positive lysozyme-staining cells as well as a few lymphocytes bearing IgG, IgA, or IgM.

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.