Shunsuke Kumasa

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 Keratin Proteins in Sweat Gland Tumours by the Use of Monoclonal Antibody

A total of 34 cases (eccrine poroma: 2, cylindroma: 2, eccrine spiradenoma: 4, syringocystadenoma papilliferum: 1, hydroadenoma papilliferum: 1, clear cell hydroadenoma: 7, mixed tumour: 16) of sweat gland tumours of the skin were described in terms of immunohistochemical distribution of keratins using polyclonal anti-keratin antiserum (TK, detecting 41-56 KDa keratins) and monoclonal antibodies (KL1, 55-57 KDa; PKK1, 40, 45, 52.5 KDa). Keratin expression in eccrine poroma, spiradenoma and syringocystadenoma was similar to that in the ductal segment of normal sweat glands. Cylindroma showed usually slight staining for kertins. Tumour cells of hydroadenomas showed not so prominent staining for any of the keratins; however, histologically, tumour cells indicated marked variation, and the degree of keratin proteins also was different among these histological variants. Mixed tumours of the skin were strongly decorated with anti-keratin antibodies along the luminal surface cells of typical structures, while no staining occurred in outer side cells. Luminal tumour cells may be differentiated from secretory coil cells, whereas outer side cells may have a myoepithelial origin, as outer layer cells found in pleomorphic adenoma of salivary glands.

Pathologic and enzyme histochemical studies on bone formation induced by bone morphogenetic protein in mouse muscle tissue.

Bone morphogenetic protein (BMP) irreversibly induced the differentiation of mesenchymal-type cells into osteoprogenitor cells for endochondral ossification. During the process of BMP-induced differentiation in mice, 4 cell type (chondroblasts, osteoblasts, chondroclasts, and osteoclasts) were examined for phosphatase and succinate dehydrogenase using a wide range of buffers (4.0 less than or equal to pH less than or equal to 9.2). During the chondroid tissue-forming stage (1 week), chondroblast-like or osteoblast-like cells expressed phosphatase activity at 6.8 less than or equal to pH less than or equal to 9.2; chondroclast-like or osteoclast-like cells expressed phosphatase activity at 4.0 less than or equal to pH less than or equal to 5.8. However, mature chondrocytes found in hyaline cartilage expressed phosphatase activity between 6.6 less than or equal to pH less than or equal to 7.6 (2 weeks). During the process of endochondral ossification, alkaline phosphatase activity decreased in osteoblast-like cells with traces of acid phosphatase activity still detectable. Chondroclastic and osteoclastic giant cells were characterized by intense succinate dehydrogenase activity.

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.

Epithelial membrane antigen as a marker of human salivary gland acinar and ductal cell function

Monoclonal antibody to epithelial membrane antigen (EMA) from human milk fat globule membrane was used with the immunoperoxidase method to examine the distribution of EMA in normal human salivary glands and in their obstructive lesions. EMA staining in the normal salivary gland was limited to luminal and lateral borders of acinar cells. The antigen was also present in high concentration on the luminal side of ductal cells. In obstructive lesions or sialoadenitis, lateral border and luminal positive staining in atrophic acinar cells was reduced or absent, and the EMA staining in duct-like structures was also markedly decreased on even absent in chronic stage. The degree of reduction in EMA staining was dependent on the extent of degeneration. Expression of immunohistochemically detectable EMA in paraffin sections suggests the antigen to be useful as a new marker of salivary gland function, of either acinar compartments or ductal segments.

Calcifying epithelioma of Malherbe with ossification. Special reference to lectin binding and immunohistochemistry of ossified sites

Lectin binding sites and immunohistochemically detectable fibronectin and carbonic anhydrase II (CA II) were examined in induced bone structure of calcifying epithelioma of Malherbe. Pathological features of ossification were particularly evident in the border zone between tumor epithelium and stroma, and epithelial foci adjacent to sites of bone formation were conspicuously basophilic. The use of lectin binding in these foci coincided with the basophilic epithelial zones: Con A, RCA‐1, PNA, SBA, and WGA bound strongly, indicating thepresence of glucose (Glu), mannose (Man), galactose (Gal), N‐acetyl‐D‐galactosamine (GalNAC), and N‐acetyl‐D‐glucosamine (GlcNAC). Fibronectin was also detected in the same epithclial‐mesenchymal interacting layers as the positive lectin‐binding sites. Staining for CA II was strongly positive in giant cells and in epithelial zones in the bone‐inducting areas. The bond induction mechanism in the stromal tissue of calicifying epithelioma of Malherbe may initially involve the action CA II in the epithelial‐mesenchymal interacting zone, which then brings about ossification of the matrix which is fibronectin‐positive and rich in lectin‐binding sites.

Immunohistochemical and Enzyme Histochemical Studies in Bone Formation Induced by Bone Morphogenetic Protein (BMP) in Mouse Muscle Tissue

It has been a well-known phenomenon that bone morphogenetic protein (BMP) heterotopically induces bone formation by causing the differentiation of osteoprogenitor cells from mesenchymal cells [1,2]. Two hypotheses of the cell origination and mechanism of bone formation by BMP have been proposed: EMP irreversibly induces the differentiation of perivascular mesenchymal cells into osteoprogenitor cells [3,4]; BMP is capable of stimulating the differentiation of skeletal muscle into cartilage [5–7]. The biological roles of bone formation induced by BMP still remain unclear. In the presnt study, the process of chondro-osteogenesis and cell differentiation for endochondral ossification by BMP implanted into mouse skeletal muscle tissue, were evaluated in terms of the immunohistochemical method for laminin, fibronectin, glycosaminoglycan, and S-100 protein, and the enzyme histochemical method of phosphatase activity with wide range of buffers (pH 4.0 to pH 9.2).