Shinichiro Sumitomo

Involucrin expression in epithelial tumors of oral and pharyngeal mucosa and skin

Involucrin has been recognized recently as a marker of terminal differentiation of squamous epithelial cells and also as a useful marker for keratinization; its expression in epithelial tumors of oral and pharyngeal mucosa and skin was examined. Involucrin in normal oral mucosa and skin was restricted to the granular and upper spinous layers and was absent in the basal layer. Hyperkeratosis was characterized by strong positive staining for involucrum in spinous and granular cell layers. A similar pattern was noted in seborrheic keratosis and verruca vulgaris. Condyloma acuminatum specimens revealed slight staining, whereas Paget cells were negative. Calcifying epitheliomas of Malherbe were usually unreactive. Papillomas exhibited the regular distribution of involucrin, as found in normal squamous epithelium. Basal cell carcinomas were generally negative, whereas squamous cell carcinomas showed an irregular distribution of involucrin. Immunohistochemical staining for involucrin may be useful for identification of keratinizing cells in epithelial tumor foci, just as is the use of monoclonal antibody to keratin KL1.

Immunolocalization of keratins in salivary gland pleomorphic adenoma using monoclonal antibodies

Monoclonal antibodies to keratins (KL1 and PKK1) were tested on paraffin sections in forty cases of pleomorphic adenoma from salivary glands. Squamous cells in the epidermis were positive for KL1, except for the basal cells, which were positive for PKK1. In normal salivary glands, ductal cells were positive for KL1 and PKK1 keratins, whereas KL1 staining of the large excretory ducts was weaker than that of PKK1. Myoepithelial cells were positive only for KL1. In pleomorphic adenoma, the following reactions were noted: a positive staining for KL1 and PKK1 was present in the inner layer of cells within tubular and ductlike structures, a markedly positive staining for KL1 was present in squamous metaplastic cells, and a negative staining reaction with KL1 and PKK1 in spindle-shaped tumor cells was observed within the outer layer. Spindle-shaped or fibroblast-like cells in the stroma, probably myoepithelial in origin, showed an irregular staining expression for KL1 and PKK1.

Comparison of CEA distribution in lesions and tumors of salivary glands as determined with monoclonal and polyclonal antibodies.

SummaryImmunohistochemical localization of carcinoembryonic antigen (CEA) with conventional antibody to CEA (anti-CEA), nonspecific crossreacting antigen (NCA)-absorbed polyclonal antibody to CEA (NCAa-CEA), and monoclonal antibody to CEA (Mono-CEA) have been compared in obstructive lesions and salivary gland tumors. Normal salivary glands gave strong staining of the luminal borders of acinar cells with anti-CEA, whereas no staining occurred with Mono-CEA. Obstructive lesions showed occasionally marked staining with anti-CEA in some acinar cells, but there was no reaction with Mono-CEA. Of 69 pleomorphic adenomas examined, 34 were positively stained with anti-CEA, 18 with NCAa-CEA and 8 with Mono-CEA along the luminal borders of the tumor cells. The frequency of positive staining of material within tubular lumina was similar with all three immunoreagents. Neoplastic cells were positive with Mono-CEA in only three cases, while eight cases were positive with NCAa-CEA and 11 cases with anti-CEA. In salivary gland tumors true CEA is be found mainly at the border of tumor cells, but the frequency of positive reactions is low.

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.

Characterization of cells in salivary gland lesions by immunohistochemical identification of carcinoembryonic antigens

Immunohistochemical demonstration of carcinoembryonic antigen (CEA) was reported in normal salivary glands and in pathologic lesions. Staining patterns of CEA and nonspecific cross-reacting antigen-absorbed CEA (NCAa-CEA) were compared. Normal salivary glands disclosed positive staining by CEA on border and luminal sides of acinar cells and occasionally in components secreted into ductal spaces with both antigens used. Chronic obstructive lesions displayed an intense CEA staining in ductlike structures and in material secreted into their lumina in the two antigens used. Pleomorphic adenoma exhibited varying intensities of CEA in neoplastic epithelial cells of ductal structures. In contrast, they showed a slight staining reaction to NCAa-CEA. Squamous-cell carcinomas showed a strong CEA reaction, whereas they showed no reaction or a trace reaction to NCAa-CEA. Positive staining to CEA in squamous neoplastic lesions was related to nonspecific reacting antigens.

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 bone morphogenetic protein (BMP) in mixed tumor of skin

Immunohistochemical reaction of bone morphogenetic protein (BMP) was assessed in 19 cases of skin mixed tumor and 5 cases of skin appendage tumors by using monoclonal antibody raised against BMP. All cases of skin mixed tumor showed positive staining for BMP in modified myoepithelial cells located at the periphery of tubulo-ductal or solid structures, and in plasmacytoid or tumor cells in hyalinous structures. Chondroidally changed cells also showed a strong BMP immunoreactivity. Tumors originating from sweat glands were devoid of BMP immunoreactivity. It is concluded that BMP is synthesized and produced in modified or transformed myoepithelial cells in skin mixed tumor and participates in the process of chondroid changes in the tumor.

Comparison of proliferating cell nuclear antigen index in benign and malignant salivary pleomorphic adenoma

The expression of proliferating cell nuclear antigen (PCNA) was studied in benign and malignant pleomorphic adenomas by using monoclonal antibody to PCNA. Carcinoma in pleomorphic adenoma (n = 8), cell-rich variant (n = 6) and typical pleomorphic adenoma (n = 6) were selected in this study. The PCNA index in carcinoma in pleomorphic adenoma showed a higher index of nuclear staining (mean 22.9%, S.D. 6.2) than in typical pleomorphic adenoma (mean 6.9%, S.D. 3.4) or a cell-rich variant of pleomorphic adenoma (mean 8.8%, S.D. 3.3). A significant difference in PCNA index was found between benign and malignant pleomorphic adenoma (P < 0.05). The present study suggests that PCNA index significantly differs between pleomorphic adenoma and carcinoma in pleomorphic adenoma, but in the prediction of malignant transformation potential it should be combined with routine histopathological examination.

Immunohistochemical study of fibroblast growth factor-2 (FGF-2) and fibroblast growth factor receptor (FGF-R) in experimental squamous cell carcinoma of rat submandibular gland.

Fibroblast growth factor-2 (FGF-2) is a member of the heparin-binding growth factor family and has mitogenic activity. Immunohistochemical expression of FGF-2 and its receptor (FGFR) was evaluated in experimentally induced squamous cell carcinoma as well as transforming cells of rat submandibular gland (SMG) induced by 9,10-dimethyl-1,2-benzanthracene (DMBA)/sponge implantation. Proliferating cells detected by proliferating cell nuclear antigen (PCNA) staining during carcinogenesis were also compared with FGF-2 and FGFR stainings. In the normal SMG, FGF-2 and FGFR were present in the excretory, striated and intercalated duct cells. Pillar and transition cells of granular convoluted tubule (GCT) showed FGF-2 staining, PCNA-labeled cells in normal SMG were rarely observed. In 2-3 weeks after carcinogen implantation, the reactions of FGF-2 and FGFR were expressed in epithelial islands, duct-like structures and affected ductal segments. PCNA-positive cells were developed in these epithelial structures. In 4-8 weeks after carcinogen implantation squamous epithelium appeared surrounding DMBA/sponge and gradually transforming with high PCNA labeling in the based cells and strong staining of FGF-2 and FGFR. Squamous cell carcinoma arose within about 12 weeks of the experiment. In squamous cell carcinoma, there was an intense immunohistochemical expression of FGF-2 and FGFR. Basal and parabasal layers of the squamous cell carcinoma showed high PCNA labeling. FGF-2-positive cells were found in the connective tissue stroma and in inflammatory cells around the proliferating duct-like structure. Coexpression of FGF-2 and FGFR was indicated in transforming cells during carcinogenic processes and in experimental squamous cell carcinoma of rat SMG. These findings suggested that FGF may play an important role for squamous metaplasia and carcinogenesis in rat SMG as an autocrine factor and FGF-positive stromal cells may also act to stimulate epithelial proliferation.

Immunohistochemical localization of carbonic anhydrase I and II in submandibular salivary glands of the mouse, rat, hamster and guinea pig

Tissues were fixed in Bouin and Carnoy solutions, and the peroxidase, anti-peroxidase (PAP) method was used. Serous acinar cells of the guinea pig were positive for carbonic anhydrase (CA), but acinar cells of rodents were negative. Granular convoluted tubule (GCT) cells of the mouse, rat and hamster had varying degrees of staining for CA isoenzyme I and II. Striated and excretory ducts from all four species were usually positive for CA. Scattered cells containing a high concentration of CA I and CA II were present in striated ducts and GCT of the hamster, GCT of the rat and striated and excretory ducts of the guinea pig.