Miyako Chanoki

Increased expression of lysyl oxidase in skin with scleroderma

Summary Lysyl oxidase initiates cross‐linkage of collagen and elastin by catalysing the formation of a lysine‐derived aldehyde. In order to study cross‐linking in scleroderma, we used monoclonal antibodies to lysyl oxidase to determine the localization of this enzyme in systemic and localized scleroderma, and compared the distributions obtained with that in normal skin. Using an indirect immunofluorescent antibody method and an avidin‐biotinylated enzyme complex method. 11 cases of diffuse type of systemic scleroderma and seven cases of localized scleroderma were studied. In the oedematous stage of systemic scleroderma, intracellular and extracellular lysyl oxidase were remarkably increased in the dermis, particularly in groups around blood vessels. In the sclerotic stage of systemic scleroderma, lysyl oxidase was detected intracellularly in fibroblasts and extracellularly among collagen bundles between the lower dermis and the subcutaneous fat tissue. In localized scleroderma, a marked increase in lysyl oxidase was observed in mononudear cells and libroblasts near blood vessels in the lower dermis and in the subcutaneous fat tissue, in addition to the extracellular deposits between collagen bundles. The increase in lysyl oxidase in localized scleroderma was much more common than in the oedematous stage of systemic scleroderma. These findings indicated that intracellular and extracellular expression of lysyl oxidase expression was greater in sclerodermatous skin than in normal skin.

Immunohistochemical localization of lysyl oxidase in normal human skin.

Lysyl oxidase (EC 1.4.3.13), a copper‐dependent enzyme which catalyses the formation of aldehyde cross‐links, and acts primarily on collagen and elastin, is known to be increased during wound healing and in fibrotic disorders including liver cirrhosis and atherosclerosis, and to be decreased in some hereditary connective tissue diseases and in malignant cell lines. A recent study showed that lysyl oxidase might possess tumour suppressor activity as an antioncogene for ras. Little is known about the localization of this enzyme in human skin. In this study, we determined immunohistochemically the localization of lysyl oxidase in normal skin of young and elderly subjects obtained from sun‐exposed and unexposed regions of the body. All skin samples tested had similar distributions of lysyl oxidase. The enzyme was present both extracellularly and intracellularly. Extracellularly, a few granular aggregates of immunoreactants were observed along collagen and elastic fibres. These granules were more common in the adventitial portion of the dermis than in the reticular portion. Of all sun‐exposed and unexposed regions studied, the skin of the face displayed the greatest amount of extracellular immunoreactants. Immunopositive granules were observed intracellularly in fibroblasts, vascular endothelial cells, sweat glands, sebaceous glands, arrector pili muscles and some keratinocytes. These findings provide evidence that, as suggested in recent reports, lysyl oxidase may have a variety of intracellular functions.

Chédiak‐Higashi Syndrome: Report of a Case and Review of the Japanese Literature

We report the case of a Japanese female infant with Chédiak‐Higashi syndrome born to consanguineous parents. At birth she had fair skin but, when she was three months old, marked hyperpigmentation of the sun‐exposed skin areas developed. Microscopic examination of blood and electron microscopic examination confirmed the diagnosis. She enjoyed good health until she was two years old when she had pneumonia with marked hepatosplenomegaly. It is important for dermatologists and pediatricians to be aware of the skin manifestations of this disease because hyperpigmentation after sun exposure may be a characteristic, initial feature of this condition.

Immunohistochemical localization of type I, III, IV, V, and VI collagens and laminin in neurofibroma and neurofibrosarcoma

By using antibodies to type I, III, IV, V, and VI collagens and laminin, we examined the localization of interstitial collagens and basement membrane components with indirect immunofluorescence and immunoelectron microscopy (IEM). Furthermore, the morphological changes of these collagens in malignant transformation were investigated. In neurofibroma. IEM showed type I, III, and V collagens to be present diffusely on cross-striated collagen fibrils, whereas type VI collagen was present between the fibrils. Type IV collagen and laminin were observed to surround tumor cells. In neurofibrosarcoma, tumor collagen bundles that reacted with antibodies to type I, III, V, and VI collagens were irregularly arranged. Immunofluorescent deposits that reacted with anti-type IV collagen and anti-laminin antibodies were decreased in number, showing a thin and sparse arrangement.

Primary cutaneous adenoid cystic carcinoma: ultrastructural study and immunolocalization of Types I, III, IV, V collagens and laminin

A primary adenoid cystic carcinoma of the skin is reported. Light microscopy revealed pseudocysts. PAS‐positive basement membrane and true glandular lumen, which in aggregates are specific for adenoid cystic carcinoma. Perineural invasion was also observed. Ultrastructural examinations revealed three types of cystic spaces; pseudocysts, true glandular lumens and intercellular spaces. Enzyme histochemical examinations showed positive reactions for eccrine enzymes, including phosphorylase and succinic dehydrogenase and negative for apocrine enzymes. Immunolocalization of collagens and laminin revealed that basement membranes of the pseudocysts involve Type V collagen as well as Type IV collagen and laminin.

Farber's lipogranulomatosis in siblings: light and electron microscopic studies

Two cases of Farbers lipogranulomatosis in siblings are reported. The clinical features included contractures of the limbs with swelling of the joints and subcutaneous nodules and erythematous infiltrated plaques. On histology there were many large foam cells in the dermis, and electron microscopy showed numerous large cells with round cytoplasmic lamellar and microtubular bodies.

Immunohistochemical localization of type V collagen in normal human skin

SummaryTissue distribution of type V collagen in normal human skin was studied using an indirect immunofluorescent technique to determine whether type V collagen is present in the interstitium or in the basement membrane. Type V collagen was isolated from the human placenta by pepsin digestion and was purified with fractioning salt precipitations. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that type V collagen contained α1(V) and α2(V) chains, but not the α3(V) chain. Specificity of the rabbit antibodies to type V collagen was assessed using enzyme-linked immunosorbent assay (ELISA) and an immunoblotting method. Antibodies showed no cross-reactivity to other collagens, laminin, and fibronectin. With an indirect immunofluorescent technique, type V collagen was found to be widely distributed throughout the dermis. Intense fluorescent staining was noted in the papillary dermis and adnexal dermis surrounding hair follicles and eccrine glands. The basement membrane of the dermoepidermal junction, skin appendages, and capillaries was not stained. By indirect immunoperoxidase double staining, type V collagen was not found to be deposited on type IV collagen present in the basement membrane. Immunoelectron microscopic studies showed that type V collagen was not located in the basal lamina. These results suggest that type V collagen is distributed in the interstitium, but not in the basement membrane of normal human skin.

PROGRESSIVE SYSTEMIC SCLEROSIS ASSOCIATED WITH CUTANEOUS AMYLOIDOSIS

A 43‐year‐old woman presented with a 3‐year history of Raynauds phenomenon and a six‐month history of numbness in both arms. Sclerosis was noted on the entire body surface. The skin of the face was smooth and the lips were constricted (Fig. 1). The fingers and hands were atrophic and sclerotic, and full extension of the fingers and metacarpal joints was impossible (Fig. 2). There was pigmentation on the dorsal aspect of the hands. From the nape to the upper back, pruritic wavy, rippled or reticular pigmented macules in addition to sclerosis were noted (Fig. 3). Other parts of her skin did not show such a wavy pigmentation. Physical examination revealed no specific findings in the lung, heart, and abdomen. Neurologic examination was unremarkable. Motor function including muscle tonus was normal.

The Subbasement Membrane Distribution of Type IV Collagen in Normal Human Skin

Samples of normal human skin were obtained from 48 sites in 26 subjects ranging in age from 2 to 85 years. The samples were examined by indirect immunofluorescence and immunoelectron microscopy using anti‐human type IV collagen antibodies produced by immunizing rabbits with type IV collagen extracted from human placenta. Fluorescence was observed as granular or fine fibrous patterns, not only in the basement membrane at the dermo‐epidermal junction, around the vessels, and the accessory organs of the skin, but also in the dermal regions in the vicinity of the basement membranes. This suggests the presence of type IV collagen in the dermis deep to the basement membrane. Ultrastructurally, the extrabasal lamina distribution of type IV collagen was noted as a partial distribution around the fibroblasts that existed close to the basal lamina. These findings are considered to be important in examining the function of this collagen in the dermis and the dynamics and metabolism of the basement membrane under normal and abnormal conditions.

Immunohistochemical localization of lysyl oxidase in normal human skin

Lysyl oxidase (EC 1.4.3.13), a copper-dependent enzyme which catalyses the formation of aldehyde cross-links, and acts primarily on collagen and elastin, is known to be increased during wound healing and in fibrotic disorders including liver cirrhosis and atherosclerosis, and to be decreased in some hereditary connective tissue diseases and in malignant cell lines. A recent study showed that lysyl oxidase might possess tumour suppressor activity as an antioncogene for ras. Little is known about the localization of this enzyme in human skin. In this study, we determined immunohistochemically the localization of lysyl oxidase in normal skin of young and elderly subjects obtained from sun-exposed and unexposed regions of the body. All skin samples tested had similar distributions of lysyl oxidase. The enzyme was present both extracellularly and intracellularly. Extracellularly, a few granular aggregates of immunoreactants were observed along collagen and elastic fibres. These granules were more common in the adventitial portion of the dermis than in the reticular portion. Of all sun-exposed and unexposed regions studied, the skin of the face displayed the greatest amount of extracellular immunoreactants. Immunopositive granules were observed intracellularly in fibroblasts, vascular endothelial cells, sweat glands, sebaceous glands, arrector pili muscles and some keratinocytes. These findings provide evidence that, as suggested in recent reports, lysyl oxidase may have a variety of intracellular functions.