Immunohistochemical localization by monoclonal antibody of human epidermal growth factor in mixed tumours of the skin

Immunohistochemical distribution of human epidermal growth factor (hEGF) was described in 17 cases of mixed tumour of the skin with monoclonal antibody. In normal sweat glands, epithelial cells in the secretory portion and in the transitional area between secretory portion and duct showed prominent staining for hEGF. In the salivary pleomorphic adenoma type of mixed tumour of the skin, luminal tumour cells of tubular and duct-like structures gave a very characteristic hEGF staining reaction. The tumour cells showing strong staining for hEGF were scattered throughout the solid foci in this type of mixed tumour. Tubular epithelial cells in the clear cell adenoma type also displayed a positive hEGF reaction. And apocrine mixed tumours strong staining for hEGF occurred on the apical side of tubular and ductal tumour cells. In view of the immunohistochemical staining patterns for hEGF, the histologic origin of mixed tumours of the skin is suggested to be cells in the secretory portion and those in the transitional portion between secretory portion and duct of the sweat gland.     

Immunohistochemical study of carbonic anhydrase in mixed tumours and adenomas of sweat and sebaceous glands

Immunohistochemical distribution of carbonic anhydrase II (CA) in mixed tumours and adenomas of sweat gland origin and in sebaceous adenomas was demonstrated by the PAP method. Normal sweat glands, both eccrine and apocrine, clear cells of the secretory coils, and ductal epithelial cells all showed conspicuous staining for CA, and sebaceous glands were also positive. Mixed tumours of the skin indicated strongly positive staining for CA in the luminal cells of tubular and duct-like or cystic structures, while most of the other tumour cells were negative. In solid or massive foci, CA positive cells were found scattered among the cellular mass. Sebaceous adenomas were usually moderately positive for CA throughout the tumour.     

Phytohemagglutinin-binding sites in the skin. A useful histochemical marker of acrosyringium and distal portions of intradermal sweat ducts

Phytohemagglutinin (PHA)-binding sites were histochemically investigated in normal adult skin and various skin tumors using peroxidase-labeled lectin. In the normal skin, acrosyringium and distal portion of intradermal duct of eccrine and apocrine sweat glands were intensely stained with PHA. Other elements of the skin were virtually negative. Among various kinds of sweat gland tumors and other skin tumors, syringoma and eccrine porocarcinoma were intensely stained with PHA. Small ductal structures of nodular hidradenoma and mixed tumor of the skin were also stained with PHA. No other skin tumors were positive with PHA staining. The present study clearly showed that PHA is a useful histochemical marker of distal portion of eccrine and apocrine sweat ducts.     

Differentiation-associated localization of small prolne-rich rotein in normal and diseased human skin

Summary The expression of SPRR (small proline-rich protein) was investigated in normal human skin and in diseased skin from patients with psoriasis, squamous cell carcinoma, basal cell epithelioma. Naevus pigmentosus, ichthyosis vulgaris and several inflammatory skin diseases, by immunohistochemical staining. A polyclonal antibody was raised against a synthetic peptide for a C-terminal common region for SPRR l and SPRR 3. In immunoblot analysis, a positive band of 18kDa was detected, which showed the presence of SPRR l in human epidermal keratinocytes. In normal epidermis, positive staining for SPRK was observed in keratinocytes in the granular layer and the uppermost or two spinous cell layers, with no staining of the other spinous or basal layers. The staining was obvious at the cell periphery, weak at the cytoplasm, and absent in the nucleus. Staining was observed in several outer layers of the follicular infundibulum to the isthmus. No staining was detected in the inner root sheath of the hair follicles, hair matrix, sebaceous gland, eccrine gland, eccrine duct, melanocytes. Langerhans cells or fibroblasts. The arrectores pilorum, striated muscles, muscle layers of vessels, and myoepithelia of eccrine gland, were weakly stained. In psoriatic skin, stained keratinocytes were distributed in the spinous cell layers except for the basal layer, in ichthyosis vulgaris. SPRR was barely expressed in the uppermost living cell layers of the epidermis in epidermolytic hyperkeratosis. degenerated squamous cells widely expressed SPRR. In Darier's disease, dyskeratolic cells were clearly stained. In squamous cell carcinoma, staining was observed in keratotic cells around horny pearls. In basal cell epithelioma, naevus pigmentosus, and malignant melanoma, the tumour cells or naevus cells were not stained. The distribution of SPRR was similar to that of involucrin in normal and several diseased skin, except for ichthyosis vulgaris. We conclude that SPRR is expressed in close association with epidermal differentiation in normal skin and skin diseases. The alteration of the expression of the proteins correlated to terminal differentiation, and differs from disease to disease.     

Two cases of clear cell acanthoma: an immunohistochemical study

Two cases of clear cell acanthoma are reported. The expression of carcinoembryonic antigen (CEA), involucrin and keratin proteins in the tumors was investigated immunohistochemically. In 1981, Penneys et al. reported that this tumor was not of sweat gland origin because of the absence of CEA. This study confirmed this, furthor, the pattern of positive reaction of involucrin also indicated that this tumor is not of swcat duct origin.     

An investigation of cytokeratin expression in skin epithelial cysts and some uncommon types of cystic tumours using chain-specific antibodies

Summary The differentiation state of skin epithelial cysts and some uncommon types of epithelial skin tumours was investigated by immunohistochemical staining, mainly using cytokeratin (CK) polypeptide-specific monoclonal antibodies. Samples of interfollicular epidermis, hair follicles and eccrine sweat glands were included as reference tissues. The CK reactivity in epidermoid cysts and milia is not restricted to CKs involved in epidermal-type differentiation, i.e. CK1, 5, 10 and 14, but in addition CK16, a hyperproliferative keratinocyte marker is suprabasally expressed. CK1 and 10 are other prominent suprabasal markers, while CK14 seems to be preferentially expressed in the basal cell layer. Of the non-epidermal CKs, only CK4 was focally or more extensively detected in about 50% of the cases. In terms of CK reactivity, keratinization of trichilemmal cysts corresponds to the keratinization of the anagen-phase hair follicle in the isthmus. The CK reactivity is again restricted to CK1, 5, 10, 14 and 16. However, the CK1 as well as CK10 reactivity is subject to serious limitations, since both CKs were only convincingly observed in foci of terminal differentiation. Eccrine hydrocystoma obligatorily expresses a complex CK set, including CK7, 8, 14, 18 and 19. This CK set perfectly corresponds to the CK composition observed in acini of eccrine sweat glands. In addition, a discontinuous CK4 and 16 reactivity was seen in about 50% of the sites, while CK1 and 10 displayed a strictly focal appearance. On the other hand, syringoma produces in its distinct structures, a CK pattern reminiscent of the one observed in eccrine sweat gland ducts and includes CK1, 5, 10, 14, 16 and 19. Finally, the CK expression pattern of pilomatricoma includes CK1, 8, 10, 14 and 19, and is reminiscent of the CK staining of hair bulb matrix cells differentiating in the keratogenous zone in the direction of hair cortex. The reactivity of CK1 and 10 was mainly restricted to foci of squamoid differentiation and also to transitional cells bordering on shadow cells, as far as it concerns CK10. Occasionally, CK7 and 16 were observed in individual cells or small cell groups. In our view these CK reactivity patterns are useful to judge the differentiation state reached in pathological conditions, but so far do not allow us unequivocally to determine the site of origin of these lesions.     

Immunohistochemical distribution of adult T-cell leukemia-derived factor/thioredoxin in epithelial components of normal and pathologic human skin conditions.

Tissues from normal human skin and various skin diseases were studied with the immunoperoxidase technique using an antibody to adult T-cell leukemia-derived factor (ADF), a homologue of human thioredoxin. Normal human skin showed positive immunostaining for ADF/thioredoxin in the outer root sheath of hair follicle, sebaceous glands, and secreting components of apocrine and eccrine sweat units, but not in the unexposed interfollicular epidermis and other parts of both hair follicles and the sweat units. Immunoreactivity of benign skin tumors gave similar distribution to their normal counterparts; trichilemmal cyst, nevus sebaceus, senile sebaceous hyperplasia, and mixed cell tumor were positive for immunostaining, whereas epidermal cyst and pilomatricoma were not. No immunoreactivity was detected in malignant skin tumors such as basal cell carcinoma and poorly differentiated squamous cell carcinoma. Solar keratosis, well-differentiated squamous cell carcinoma, some of metastatic lesions of squamous cell carcinoma, and extramammary Paget's disease reacted with the antibody. These immunoreactivities reflected numerous functions of thioredoxin in higher organisms. Our findings suggest that the expression of ADF/thioredoxin in both normal and abnormal human skin is related to epithelial cell differentiation.     

细胞角蛋白在皮肤附属器肿瘤的表达及其诊断意义

目的:观察7种细胞角蛋白(cytokeratins,CK)在皮肤附属器肿瘤的表达,并探讨其意义。方法:应用免疫组织化学S-P法对49例不同皮肤附属器肿瘤进行CK7(K72.2)、CK8(C-51)、CK10(DE-K10)、CK14(LL002)、CK17(E3)、CK18(DC10)、CK19(KS19.1)标记,观察不同细胞角蛋白的表达和分布模式。结果:49例皮肤附属器肿瘤中7种细胞角蛋白的表达和分布有所不同。毛发分化的肿瘤中CK表达多数较弥漫;而汗腺分化的肿瘤中,不同CK表达位置明显不同,每种肿瘤各有特点。结论:综合分析CK1018表达情况,可能有助于汗腺肿瘤与毛囊肿瘤的鉴别,CK10-18 支持为汗腺肿瘤,CK10 18-支持为毛囊肿瘤。     

七种细胞角蛋白在皮肤上皮性肿瘤中的表达

目的 观察7种细胞角蛋白在皮肤上皮性肿瘤中的表达,并探讨其意义.方法 应用免疫组化S-P法对54例不同的皮肤上皮性肿瘤和20例正常皮肤进行细胞角蛋白7(K72.2)、细胞角蛋白8(C-51)、细胞角蛋白10(DE-K10)、细胞角蛋白14(LL002)、细胞角蛋白17(E3)、细胞角蛋白18(DC10)、细胞角蛋白19(KS19.1)标记,观察不同细胞角蛋白的表达.结果 54例皮肤上皮性肿瘤包括,鳞状细胞癌10例、基底细胞癌10例、毛发肿瘤19例、皮脂腺癌2例、汗腺肿瘤13例.皮肤上皮性肿瘤中7种细胞角蛋白的表达和分布有所不同.鳞状细胞癌、基底细胞癌和毛发分化的肿瘤中细胞角蛋白多数呈弥漫表达;而汗腺分化的肿瘤中,不同部位表达不同细胞角蛋白,每种肿瘤各有特点.结论 选择地检测一组细胞角蛋白的联合表达,有助于皮肤上皮性肿瘤的诊断和鉴别诊断.     

Hedgehog pathway does not play a role in hidradenitis suppurativa pathogenesis

Hidradenitis suppurativa is a chronically relapsing skin disorder with onset after puberty and is characterized by inflammatory lesions in hair follicle and apocrine sweat gland-bearing skin that manifests as abscesses with formation of cysts and sinus tracts. Hedgehog family genes are required in normal embryonic skin, hair follicle, sebaceous and sweat gland development. Mutations of hedgehog pathway in adult skin have previously been found in basal cell carcinomas and in alopecia as well as in epidermal cysts and in odontogenic keratocysts. Therefore, we suggested that the hedgehog pathway might play a role in formation of sinus tracts and cysts as newly formed structures in hidradenitis suppurativa patients. None of the sinus tracts or cysts in 81 hidradenitis suppurativa histological slides from 34 patients showed positive finding for sonic hedgehog mutation. According to our findings, we have to conclude that there is no evidence that sonic hedgehog pathway is part of hidradenitis suppurativa pathogenesis.     

Distribution of metallothionein in normal and pathological human skin

The expression and distribution of metallothionein (MT) in frozen sections of normal and pathological human skin was studied using the monoclonal antibody L2E3 directed against MT derived from human fetal liver. Immunohistochemical staining of normal fetal and adult skin revealed strong reactivity in basal keratinocytes of epidermis and outer hair root sheath, hair matrix cells and the secretory coil, but not the exocrine portion of eccrine glands; myoepithelial cells around apocrine sweat glands were similarly stained. In epidermal hyperplasia, variable numbers of suprabasal keratinocytes were stained, whereas in interface dermatitis, interrupted staining was found in the basal layer. Weak or scattered staining was observed in squamous tumours, whereas basal cell carcinomas did not show consistent staining. The distribution of MT in normal skin was in line with the germinative role of basal keratinocytes and hair matrix cells, whereas its distribution in hyperplastic epidermis was in line with experimental animal data, and reflected the increase in the germinative pool in these conditions. It is concluded that monoclonal antibody L2E3 may serve as a valuable immunohistochemical marker in diagnostic cutaneous pathology since it labels basal keratinocytes selectively, and since it discriminates between eccrine and apocrine sweat glands.     

Immunohistochemical demonstration of carcinoembryonic antigen and related antigens in various cutaneous keratinous neoplasms and verruca vulgaris

Carcinoembryonic antigen (CEA), which is a well-known marker for the normal sweat gland apparatus and its neoplasms in the skin, was recently demonstrated in sebaceous neoplasms. The aim of this study was to examine the expression of CEA and related antigens in the other cutaneous keratinous neoplasms and verruca vulgaris. Normal adult skin, squamous cell carcinoma (SCC), senile keratosis, Bowen's disease, basal cell carcinoma (BCC), seborrhoeic keratosis and verruca vulgaris were stained immunohistochemically with a panel of monoclonal and polyclonal antibodies that recognize different epitopes of CEA and related molecules. Localization of the antigens was compared with that of involucrin and proliferating cell nuclear antigen. The strongest expression of CEA-related antigens, other than non-specific cross-reacting antigen (NCA) -50/90, was seen in SCC and verruca vulgaris, while no detectable expression was seen in BCC. Senile keratosis, Bowen's disease and seborrhoeic keratosis showed the predominance of the CEA-related antigens over CEA weakly expressed. Strong expression of both CEA and NCA-50/90 was seen only in SCC. All the expressions were limited to the cells situated in the upper epidermal cell layers of the tumours, at the centre of tumour islands in SCC and along the pseudohorn cysts in seborrhoeic keratosis, where involucrin was coexpressed. We suggest that CEA and related antigens are not only markers for sweat gland differentiation in the skin, as currently accepted, but are also expressed in various cutaneous keratinous neoplasms and verruca vulgaris. The expression may correlate with the terminal differentiation of the tumour cells, the strong coexpression of CEA and NCA-50/90 may correlate with the malignant potential of the tumour types, and the mechanisms that control the expression of CEA and related antigens in the neoplasms may be similar to those operative in verruca vulgaris.     

Activity of 17β-hydroxysteroid dehydrogenase in various tissues of human skin

The activity of 17β-hydroxysteroid dehydrogenase (17β-HSD) was assayed in various tissues microdissected from the freeze-dried human skin of fourteen subjects. The apocrine sweat gland, sebaceous gland and hair follicle possessed a high activity of 17β-HSD. The enzyme activity was negligible in the epidermis, except that the scalp epidermis showed much the same activity as the hair follicle. The dermis showed variable activity because of contamination with other components.     

Immunohistochemical demonstration of ferritin in sweat gland and sweat gland neoplasms

Using a rabbit anti-human liver ferritin antibody, we examined the binding patterns of this reagent in normal skin and observed a unique binding pattern limited to the outermost layer of the eccrine duct. Examination of a variety of sweat gland neoplasms revealed 2 distinct patterns. One was the binding of this antibody to the outermost layer of cells in the epithelial cords of syringoma, producing a characteristic ring when seen in cross-section. This pattern of binding did not occur in other neoplasms known to be related to the eccrine duct such as dermal duct tumor and eccrine poroma. Only sparse sporadic binding occurred in other eccrine and apocrine neoplasms. A second characteristic binding pattern, not related to that noted in syringoma and diffuse in pattern, was seen in acrospiroma and in a number of adnexal carcinomas. Diffuse ferritin expression has been described in malignant neoplasms in tissues other than skin. Diffuse ferritin staining of certain sweat gland neoplasms may be an indication of biologic activity and potential aggressivity of these neoplasms.     

Immunohistochemical markers of sweat gland tumors

Using immunoperoxidase methods, normal sweat glands, 44 benign and 4 malignant sweat gland tumors were tested for the presence of carcinoembryonic antigen (CEA), pregnancy-specific-B1-glycoprotein (SP1) and actin (ACT). CEA and SP1 stained the secretory and duct-lining cells of normal eccrine glands. Among benign tumors, 74% were positive for CEA and 44% for SP1. The staining reaction was found mainly in luminal secretions and surrounding cells. Staining by SP1 was reduced, but not suppressed, after absorption with the purified antigen. ACT was found in myoepithelial cells of the secretory tract of normal glands and in basal cells of all cases of hidradenoma papilliferum. Only 3 sweat gland carcinomas reacted for CEA. In a malignant chondroid syringoma, no ACT-positive cells were seen in the myxochondroid stroma. The potential value of CEA, SP1 and ACT in the diagnosis of sweat gland tumors is discussed.     

Anti-hair keratin monoclonal antibody (HKN-2)

BALB/c mice were immunized with human hair fibrous proteins. Then their spleen cells were fused with Sp2/0-Agl4 mouse myeloma cells. Antibody-producing hybridomas were selected by ELISA method and cloned by limiting dilution. A monoclonal antibody was chosen and designated as HKN-2. Immunohistochemically, on frozen sections of normal human skin, HKN-2 was found to recognize the suprabasal cells of epidermis and hair follicle, the cells in the keratogenous zone of hair shaft, the sebaceous cells and the ductal and myoepithelial cells of sweat glands. The basal cells of the epidermis and lower hair follicle, hair matrix cells and secretory cells of sweat glands did not react with HKN-2. An immunoelectron microscopic method showed that the positive reaction to HKN-2 was located on the tonofilaments in the cytoplasm. It was concluded that there might be a common antigenic determinant between hair and other skin epithelial tissues. A complexity in keratin manifestation in each epithelial tissue of the skin was suspected.     

Infundibular keratoses—a prototype of benign infundibular tumours

Three cases of a benign follicular tumour of infundibular origin are reported. The neoplasms were solitary, verrucous, slowly-growing papules or nodules on the face, which were diagnosed clinically as verruca vulgaris or seborrhoeic keratoses. Histologically, several epithelial lobules were seen, mainly above the level of the surface of the surrounding skin, with characteristic funnel-shaped invaginations. The tumours occasionally contained vellus hairs or were connected with sebaceous glands and/or hair follicles at their bases, indicating their follicular origin. The tumour masses consisted of peripherally arranged basaloid and inner squamoid cells. The latter cells contained more glycogen and appeared paler with haematoxylin and eosin (H&E) stains than the normal inter-follicular squamous cells. Neither clear cells nor squamous eddies were observed. Palisading of the basaloid cells was not a prominent feature. The name 'infundibular keratosis' is proposed for such tumours, which probably represent the prototype of infundibular tumours of the hair follicle.     

Eccrine sweat glands associate with the human hair follicle within a defined compartment of dermal white adipose tissue

It has always been assumed that sweat glands are distributed throughout the skin unconnected with hair follicles (hair “roots”) and their associated sebaceous (or “grease”) glands, which, together with other specialised glands and the muscles that make hairs stand up, are known as pilosebaceous units. Sweat glands and pilosebaceous units, in other words, have always been considered separate structures, with sweat glands opening on to the skin surface between hairs rather than alongside them. Sweat glands on the palms of the hands and the soles of the feet develop differently and were not considered in this study. Using a number of techniques, including looking down the microscope at sections of hair follicles taken from the scalp during hair transplants, and at manual and computer‐assisted 3D reconstructions of sections of scalp skin, this study from Spain and Manchester, UK, suggests that in hair‐bearing areas the deepest part of the sweat gland, a coiled structure, is in fact located very close to the deepest part of the pilosebaceous unit, sitting right by the sheath of the hair follicle, below the sebaceous gland and the point where the hair muscle attaches. Moreover, these structures sit together within cone‐shaped projections of fat that reach upwards into the lowest layer of the skin, the thick reticular dermis, from the fat underneath. The authors speculate that sweat glands, pilosebaceous units and this particular fatty tissue (collectively the “adnexal skin unit”) may interact in certain skin diseases, during wound healing and with drug treatments.     

Lectin-binding sites in eccrine sweat gland tumours

Lectin-binding sites in sections of formalin-fixed, paraffin-embedded eccrine sweat gland tumours were investigated using fluorescein isothiocyanate (FITC) conjugated peanut agglutinin (PNA), FITC conjugated Ricinus communis 1 agglutinin (RCA-1) and FITC conjugated wheat germ agglutinin (WGA). In 22 benign eccrine sweat gland tumours, lectin-binding sites were noted primarily on the cell surface, and in the secretions. In five malignant sweat gland tumours, all showed cytoplasmic lectin-binding sites in variable proportions of malignant cells in addition to cell surface staining. These results indicate that cytoplasmic lectin-binding sites may be a useful marker of neoplastic transformation of eccrine sweat gland tumours.     

小鼠毛囊周期中Thy-1抗原的表达

目的　观察拔毛诱导的毛发生长周期中毛囊Thy 1抗原的表达。 方法　自然休止期C57BL/6小鼠 ,通过拔毛诱导其毛发自休止期进入生长期 ,应用ABC免疫组化法连续观察毛囊Thy 1抗原的表达及组织学改变。结果　拔毛后第 2天 ,漏斗部毛囊内层角质形成细胞明显表达Thy 1抗原 ,随后转为毛囊周围浸润细胞及基质较强的表达Thy 1抗原 ,以 7～ 9天最强 ,以后逐渐减弱 ,至 17～ 2 0天基本消退。结论　拔毛刺激毛发生长周期中毛囊上皮可一过性表达Thy 1抗原。毛囊上皮的这种Thy 1抗原的表达 ,在皮肤免疫应答中可能起重要的作用