Kazufumi Yoneda

Immunohistochemical staining properties of amyloids with anti-keratin antibodies using formalin-fixed, paraffin-embedded sections

Immunohistochemical staining properties of amyloids with anti‐keratin antibodies were investigated using an avidin‐biotin‐per‐oxidase complex (ABC) system on formalin‐fixed, paraffin‐embedded sections. Anti‐keratin antibody EAB‐903 which recognize 66K and 57K daltons keratin peptides reacted with amyloid deposits in both lichen amyloidosus (LA) and macular amyloidosis (MA), but did not react with either primary systemic amyloidosis (AL), secondary systemic amyloidosis (AA) or heredo‐familial amyloid polyneuropathy (AF). However, anti‐keratin antibodies EAB‐904 and MAK‐6 did not react with any types of amyloids. These results suggested that immunohistochemical staining with anti‐keratin antibody EAB‐903 using formalin‐fixed, paraffin‐embedded sections appeared to be a useful method in making differential diagnosis of primary localized cutaneous amyloidosis (AD).

Differentiation of eccrine poroma cells to cytokeratin 1- and 10-expressing cells, the intermediate layer cells of eccrine sweat duct, in the tumor cell nests.

It has been shown that an intermediate cell layer exists between a luminal cell layer and a peripheral cell layer in human eccrine sweat ducts by immunohistochemistry using anti‐keratin antibodies 34βB4 and DE‐K10. These antibodies react to cytokeratin 1 and 10 respectively, and stain the intermediate cells specifically, but not luminal cells nor peripheral cells. Cytokeratin 1 and 10 are considered to appear as a differentiated keratin in the terminal process of epidermal keratinization. We examined 5 cases of eccrine poroma with 34βB4 and DE‐K10. Various numbers of the poroid cells reacted to these anti‐keratin antibodies in 4 cases. Some positive cells were observed around the cuticular cells in two of them. The present study demonstrated that terminal differentiation in terms of keratinization can occur in eccrine poromas, and that the 34βB4‐ and DE‐K10‐positive cells around the cuticular cells differentiate toward the intermediate cells in cytokeratin expression profile and location.

Immunohistochemical study of human eccrine sweat ducts with anti-keratin antibodies : presence of a layer between luminal and peripheral cell layers

It is known that human eccrine sweat ducts are composed of luminal cells and peripheral cells. In this study, the immunohistochemical staining properties of human eccrine sweat ducts were investigated using seven different anti-keratin antibodies by light microscopy. Anti-keratin antibody MA904, which reacts with 68-kDa keratin peptide specifically stained an intermediate cell layer between the luminal cell layer and the peripheral cell layer in the ducts. Anti-keratin antibody CK8,60 stained both the luminal cell layer and the intermediate cell layer. Anti-keratin antibody MA903 stained all of the layers. Anti-keratin antibodies CK4.26, PKK1, and MAK-6 weakly to faintly stained the luminal cells. Anti-keratin antibody PKK3 stained no cells in the ducts. These results suggest that each cell layer has its own characteristic staining pattern with anti-keratin antibodies. Moreover, the presence of an intermediate cell layer was confirmed by immunoelectron microscopy using anti-keratin antibody MA904.

The distribution of keratin type intermediate‐sized filaments in so‐called mixed tumour of the skin

Epithelial cells can be distinguished from various non‐epithelial cells by the presence of keratin‐type intermediate‐sized filaments, which can be detected by immunofluorescence microscopy, using antibodies to α‐keratin. In the present study, two types of antibodies were obtained. One of them was specific for α‐keratin (mol. wt. 49,000 to 69,000 daltons) in whole epidermis, and the other for α‐keratin (mol. wt. 62,000 and 69,000 daltons) in prickle and granular cells but not in basal cells. Four cases of so‐called mixed tumour of the skin were studied by immunofluorescence microscopy using these antibodies. Tumour cell nests of cuboidal and polygonal cells, tubular structures and keratinous cysts reacted with these antibodies, as did individually‐dispersed tumour cells within the myxoid and chondroid matrix. These results indicate that all the tumour cells of the so‐called mixed tumour of the skin are of epithelial origin. Differences in staining intensity between these tumour cells and their specificity for these two antibodies are discussed with reference to keratin differentiation in tumour cells.

Immunohistochemical Staining Properties of Keratin Type Intermediate Filaments in Mammary and Extramammary Paget's Disease

Utilizing three different anti‐keratin antibodies and the avidin‐biotin peroxidase complex system on sections of formalin‐fixed paraffin‐embedded tissues and on cryostat sections, immunohistochemical localization of keratin type intermediate filaments in mammary Pagets disease and extramammary Pagets disease was investigated. Anti‐keratin antibodies EAB‐903 and EAB‐904, which recognize 66K and 57K dalton keratin peptides, did not decorate any Pagets cells in either mammary or extramammary Pagets disease. On the other hand, anti‐keratin antibody MAK‐6, which recognizes 52.5K, 50K, 48K, 45K and 40K daltons keratin peptides, did decorate Pagets cells in both Pagets diseases. These staining properties of Pagets cells were the same as those of secretory cells in normal human sweat glands and mammary glands. Anti‐keratin antibody MAK‐6 is thought to be useful in the diagnosis of mammary and extramammary Pagets diseases.

Elastic fiber and cutaneous amyloidoses

Abstract The deposition of amyloid in the skin is seen in systemic amyloidosis as well as in primary cutaneous amyloidoses, in which amyloid deposits are seen around the elastic fibers. In this chapter, we will focus on the relationship between amyloid deposits and elastic fibers.

Intercellular adhesion molecule-1 on cultured human melanoma cells: influence of cytokines.

Adhesion molecules such as intercellular adhesion molecule‐1 (ICAM‐1) and its counter‐receptor, lymphocyte function associated antigen‐1 (LFA‐1), play very important roles in immune responses. In this study, the effects of cytokines on cultured human melanoma cells (MMG2) were examined, especially focussing on the expression of ICAM‐1 on MMG2 and lymphocyte adhesion to MMG2. Both the expression of ICAM‐1 and HLA‐DR on MMG2 increased after treatment with IFN‐gamma. ICAM‐1 expression began to increase earlier than HLA‐DR expression. TNF‐alpha and IL‐1 beta also increased the expression of ICAM‐1 on MMG2. However, these cytokines did not increase the expression of HLA‐DR. IFN‐gamma had a dose dependent effect on lymphocyte adhesion to MMG2. Pretreatment of IFN‐gamma treated MMG2 with 84H10 (anti‐ICAM‐1 antibody) or pretreatment of lymphocytes with either SPV‐L7 (anti‐LFA‐1 alpha antibody) or IOT10 (anti‐LFA‐1 beta antibody) inhibited the lymphocyte adhesion to MMG2. These results suggest that ICAM‐1 molecules induced on melanoma cells by IFN‐gamma can interact with LFA‐1 molecules on lymphocytes.

A Case of Superficial Spreading Melanoma In situ 2.5 mm in Diameter

A 60‐year‐old woman with superficial spreading melanoma in situ, measuring 2.5 mm in diameter, was examined. She had noticed a very small pigmented lesion 1.2 mm in diameter on her left lower leg in April of 1989. By April of 1990, it had grown to 2.5 mm in diameter. Its edge was irregular, and its color was variegated black to brown. Skin surface markings had disappeared in the center portion. Histopathologically, the lesion was asymmetrical. Atypical large cells nested in the lower epidermis and were scattered singly in the mid and upper epidermis, as seen in Pagets disease. At the periphery of the lesion, single large tumor cells were scattered in the mid epidermis. The tumor cells reacted to monoclonal anti‐melanoma antibody HMB‐45.

Vimentin expression in sweat gland tumours

Immunohistochemical localization of vimentin was studied in 93 cases of sweat gland tumours using a monoclonal anti-vimentin antibody. A strong immunoreactivity of vimentin was observed in modified myoepithelial or neoplastic myoepithelial cells of mixed tumour of the skin, syringoma, and sweat gland adenoma. Tumour cells in outer layers of tubular, ductal, and duct-like structures usually showed positive staining for vimentin, which coincided with modified myoepithelial cells. All tumour cells of clear cell hydroadenoma showed positive vimentin staining. Tumour cells of the luminal border of tubulo-ductal structures of mixed tumours were rarely immunoreactive for vimentin. Positive vimentin staining of tumour cells in the outer zone of tubulo-ductal structures in sweat gland tumours may be related to reactive proliferation of modified myoepithelial cells and simultaneous growth of luminal tumour cells.

HETEROGENEITY OF EPIDERMAL PREKERATIN AND KERATIN BETWEEN SOLE AND BODY SKIN IN HUMANS

Recently, heterogeneity of stratum corneum keratin (α‐fibrous protein) in various anatomic sites has been described in humans as well as cows and rats. In order to study this heterogeneity more clearly in humans, we compared acid soluble (citrate buffer at pH 2.65) prekeratin, acid insoluble (8 M urea‐2‐mercaptoethanol soluble) living cell keratin, and stratum corneum keratin from sole and body (nonsole‐nonpalm) skin using SDS‐polyacrylamide gel electrophoresis (PAGE), amino acid analysis, and immunological analyses.