Studies on fibronectin in the skin

Fibronectin is a glycoprotein mediating contact between cellular elements and collagen. As judged by indirect immunofluorescence studies fibronectin is abundantly present in normal human skin. It is located in the dermo-epidermal junction area, in the papillary and reticular dermis, about epidermal appendages (pilosebaceous units and eccrine sweat glands) and in the vascular and neural structures.     

Studies on fibronectin in the skin

Fibronectin is an important constituent of normal human skin, mediating cell/cell and cell/fibre interactions. In affected skin in discoid and systemic LE, changes in the distribution of fibronectin in the dermo-epidermal junction and in the papillary dermis are observed with homogenization and degenerative changes, IF negative gaps and slit formation in the dermo-epidermal region, together with IF positive globular bodies and transport of fibronectin into the epidermis. Unaffected skin in LE demonstrates the pattern of fibronectin as found in normal human skin.     

Immunofluorescence in psoriasis: Studies of immunoglobulins,complement deposits,and thre membrane markers

Summary Sixteen psoriatic patients and 11 control subjects were investigated by immunofluorescence for skin immunoglobulins (IG) and complement (c) deposits and for keratinocyte membrane markers with anti 2 microglobulin ( 2m) antibodies (Ab), ConA, and pemphigus Ab. IG and/or c deposits were almost constant in involved epidermis. Three patterns were associated: (1) parakeratotic nuclear (dots-dashes), (2) stratum corneum (SC) intercellular (lamellar pattern), (3) vascular (granular or linear deposits in papillary dermal vessels). In uninvolved epidermis nuclear or vascular deposits could occasionnally be present but intercellular pattern was never found in SC. Control specimens were always negative.The three surface markers investigated (2 m, ConA receptor, Pemphigus antigen) could be demonstrated on psoriatic keratinocytes with only slight differences in distribution when compared with controls. Thus, by this methodology no important abnormality could be found in psoriatic keratinocyte membrane antigens.     

Distribution of carbohydrate residues in normal skin

Summary Sections of biopsies of normal skin obtained from 11 individuals were incubated with 8 lectins using an avidin-biotin complex (ABC). All sections when incubated with the appropriate lectin showed the presence of the following carbohydrate residues: l-fucose, -(1–4)-d-GlcNAc)₂ (N-acetylglucosamine), acetylneuraminic acid, Gal--(1–3)-GalNAc (N-acetyl-galactosamine), -d-galactose, -d-glucose, and -d-mannose. In addition, sections of individuals with blood group A showed -d-GalNAc and sections of individuals with blood group B showed -d-galactose. In the stratum (str.) basale, carbohydrates were present in small quantities, but as the cells matured and moved upward, the incorporation of carbohydrates into the cell membranes increased considerably. In the str. granulosum, lectin reactivity was absent in many sections, probably due to masking by phospholipids. The dark cells in the eccrine glands showed reactivity with all lectins except in the one nonsecretor with blood group A₁, whose dark cells showed no l-fucose and -d-GalNAc. The endothelial cells of the blood vessels showed lectin reactivity except when incubated with concanavalin A. The sebaceous glands showed both cytoplasmic and membrane staining when incubated with various lectins.     

银屑病患者血浆和皮损纤维连接蛋白含量测定及分子降解状态的研究

为了探讨银屑病患者血浆纤维连接蛋白（ＰＦＮ）和皮损可溶性纤维连接蛋白（ＳＦＮ）的存在状态，采用火箭电泳法和蛋白印迹法分别对９０例银屑病ＰＦＮ和２０例银屑病斑块状皮损ＳＦＮ进行检测。结果表明，患者组ＰＦＮ水平显著低于正常人对照组，皮损ＳＦＮ含量明显高于正常人皮肤。扫描结果提示，患者组ＰＦＮ降解增强可能是ＰＦＮ减少的主要原因，不同期银屑病组ＰＦＮ降解程度有差异。皮损ＳＦＮ增多可能是由于局部合成增加及ＰＦＮ向表皮渗漏所致。动态观察银屑病患者ＰＦＮ分子降解状态，具有重要的临床意义     

皮肤鳞癌中纤维连接蛋白表达对癌生物学行为的影响

观察纤维连接蛋白在皮肤鳞癌中的表现形式,探讨其与ＳＳＣ生物学行为和癌间质巨噬细胞,淋巴细胞反应的关系。用ＦＮ、ＰＣＮＡ、ＣＤ６８地５０例ＳＳＣ作免疫组化染色,检测在癌中的表达。结果显示ＦＮ在ＳＳＣ中表现为３种形式,细胞ＦＮ、基膜ＦＮ与间质ＦＮ,细胞ＦＮ与基膜ＦＮ常同时消失,与ＳＳＣ的生长、分化、增殖,转移密切相关,而且其减少程度与局部ＭφＬｃ浸润量的减少有潜在的平行关系。本文结果说明ＳＳＣ中细胞Ｆ     

Distribution of T-cell subsets as defined by monoclonal antibodies in skin lesions of psoriasis vulgaris

Summary Cryostat tissue sections from skin lesions of 16 patients suffering from chronic stationary psoriasis vulgaris were assayed for the presence of distinct T-cell subpopulations with monoclonal antibodies. Using two pan-T surface markers (M-T 4–11 and Lyt 3) the total number of infiltrating T-cells was measured. This cell population was further dissected into Leu 3a (helper/inducer) and M-T 8–11 (cytotoxic/suppressor) positive subsets. Percentages of T-cell subpopulations were within the ranges found in healthy peripheral blood and were thus regarded as normal.     

Carbonic anhydrase — a marker for fenestrated capillaries in psoriasis

Summary A strong correlation between capillary fenestrations and the demonstration of carbonic anhydrase (CA) has previously been shown. In the present histochemical study we sought to determine whether CA could serve as a marker for fenestrated capillaries in psoriasis. In normal human skin capillary staining for CA was found only in the fenestrated capillaries of the perifollicular and periglandular plexus. In psoriatic skin lesions, however, the intrapapillary capillaries also reacted for CA. From ultrastructural investigations it is known that these capillaries are fenestrated. Our findings have shown that there is a strong correlation between fenestrated and CA-positive capillaries in normal human skin as well as in the lesional skin of psoriatics. Therefore, the demonstration of CA activity may serve as a specific and sensitive marker for fenestrated capillaries in psoriasis using an uncomplicated method, which makes it possible to detect numerous fenestrated capillaries in a single histological section.     

仙方消银片对银屑病模型小鼠尾部鳞片颗粒层细胞形成的影响

目的:旨在探索仙方消银片对寻常型银屑病的疗效与作用机制。方法:采用小鼠天然尾部鳞片模型,对实验组小鼠灌胃给药仙方消银片(水溶液)0.5 mL,每日1次,连续14天,并设灌胃给药复方青黛丸(水溶液)作对照,生理盐水灌胃作空白对照,观察鼠尾鳞片颗粒层细胞数。结果:实验组鼠尾鳞片颗粒层细胞数多于对照组,差异有统计学意义(P<0.05)。结论:仙方消银片具有促使动物模型颗粒层形成的作用,提示该方在临床应用时可能纠正银屑病的表皮增生。     

Immunohistochemical staining of proliferating cell nuclear antigen (PCNA) in malignant and nonmalignant skin diseases

Abstract Immunohistochemical staining of proliferating cell nuclear antigen (PCNA) was performed in skin from patients with various malignant and nonmalignant skin diseases using anti-PCNA monoclonal antibodies. The malignant diseases included squamous cell carcinoma (SCC), adult T lymphotrophic leukemia (ATL), mycosis fungoides, malignant melanoma and malignant lymphoma, and the nonmalignant diseases included severe treatment-resistant atopic dermatitis (AD), psoriasis vulgaris, verruca vulgaris, and others. The percentage of PCNA-positive cells (the labeling index, LI) was highest for the malignant diseases (56.5 ± 7.1%). The LIs for severe treatment-resistant AD, psoriasis, and verruca vulgaris were also significantly higher than those for the normal control or nonlesional skin of the patients. The PCNA LIs were, however, not significantly elevated in eczema and contact dermatitis. The high PCNA LIs in severe AD and psoriasis vulgaris were considerably lower in the skin improved by treatment. Labeling with Ki67, a nuclear protein expressed in cycling cells, was also performed in skin from subsets of each patient group. The results were very similar to those found with PCNA labeling. PCNA-positive cells were found throughout the dermis as well as the basal layer in the malignant diseases, whereas they were found only in the basal layer in the nonmalignant diseases. The results suggest that in human skin diseases, the extent of staining for PCNA, which is a cofactor of DNA polymerase-delta and is essential for cell proliferation, correlates with the extent to which the disease is treatment-resistant. In addition, our findings suggest that the PCNA LI and distribution of PCNA-positive cells in the skin may be helpful in the early diagnosis of skin malignancies. Received: 7 July 1998 / Received after revision: 29 March 1999 / Accepted: 26 April 1999     

Intra- and inter-individual variations in cornified envelope peptide composition in normal and psoriatic skin

Summary Cornified envelopes from the stratum corneum of healthy volunteers and from the involved and uninvolved skin of psoriatic patients were electrophoretically purified, and their peptide composition analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS PAGE) after cyanogen bromide cleavage. The resulting envelope peptide patterns (EPPs) were compared. In normal subjects, mainly quantitative minor differences in the EPPs were observed between different individuals. In the same individual, palms and soles could be distinguished from other body sites by their EPPs. The palm and sole samples presented identical patterns which were different from the patterns found with samples from other body sites. In psoriatic patients, EPPs of uninvolved skin resembled closely those of healthy epidermis, but showed striking differences from those of lesional skin. The EPPs of psoriatic lesional skin showed a characteristic accumulation of small peptides with molecular weights of 3–11 kDa. The EPP of lesional skin returned to normal during PUVA therapy, indicating that the changes in the biochemical composition of the cornified envelope are correlated with the clinical status of the disease.     

Staphylococci of the normal human skin flora

Summary 352 strains of Staphylococci of the normal human skin flora were sampled from one volunteer by single scrabbing in a ca. 3 cm² measuring area. They were biotyped by the scheme of Pelzer et al. (1973)—a modified Baird-Parker-Scheme (1963)— and the resistance to antibiotics was investigated by the method of Bauer et al. (1966).All the nine biotypes of Staphylococci were found in variable quantities. It seems problematic to call one biotype as the main type. Morphologically identical colonies of Staphylococci from the indigenous flora of the human skin were not identical in their biotypes as previously described by Pelzer (1976).Only the investigation of all Staphylococci colonies from the culture plate can evaluate all biotypes of Staphylococci of the normal human skin flora, and can give the right quantitative correlation. Staphylococci were found to be sensitive and resistant up to four antibiotics, and one biotype did not show one type of antibiogram.     

寻常性银屑病患者与正常人SPRRs基因的研究

目的　探讨SPRRs基因的外显子编码区序列与银屑病发病的关系。方法　从正常人和寻常性银屑病患者的外周血中提取基因组DNA ,用自动测序的方法测定SPRRs基因的外显子编码区序列。结果　正常人和银屑病患者的SPRR2A ,SPRR2B及SPRR2D基因外显子编码区序列均相同 ;SPRR2EcDNA在 +15 6bp处存在A或G二态性 ,可表现为AA纯合、GG纯合和AG杂合三种基因型 ,但不改变蛋白质编码 ,其分布在银屑病患者和正常人之间差异无显著性。结论　SPRRs基因的外显子编码区序列与寻常性银屑病的发病无明显相关性     

Immunohistochemical localization of type V collagen in normal human skin

Summary Tissue 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 1(V) and 2(V) chains, but not the 3(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.     

Studies on fibronectin in the skin. VII. Production in cell cultures from normal human skin

In the present study, cell cultures of fibroblasts from normal skin have been investigated regarding the production of fibronectin. The development of multimeric insoluble fibronectin is demonstrated as small dots at the cell surface, developing into a branched meshwork of fibrous structures in parallel arrays. Soluble dimeric fibronectin is also found in the culture medium.