Keratinocytes In Normal Epidermis Research Articles

A new target for squamous cell skin cancer?

Prostaglandins (PGs) derived from arachidonic acid of cell membrane are synthesized by PG G/H synthase (cyclooxygenase; COX-1/-2) and signal as autocrine/paracrine lipids (1, 2). Similar to other tissues, COX-1 is constitutively expressed in keratinocytes of normal epidermis, whereas COX-2 expression is more variable and regulated (3, 4). COX-2 is induced for example by cytokines and growth factors; accumulated prostaglandins from COX-2 regulate pain, inflammation, and cancers (2, 3). In the case of cancer, increased levels of PGs disrupt differentiation and thus contribute to the sensitization of cells to carcinogens and ensuing hyperplasia (5, 6). Therefore, the inhibition of COX-2 activity with aspirin like compounds has been suggested as a potent chemo-preventive therapy to suppress tumor development, not just in skin but particularly for gastrointestinal cancers (2, 7).

Immunolocalization of the Tumor-Sensitive Calmodulin-Like Protein CALML3 in Normal Human Skin and Hyperproliferative Skin Disorders

Background and ObjectiveCalmodulin-like protein CALML3 is an epithelial-specific protein regulated during keratinocyte differentiation in vitro. CALML3 expression is downregulated in breast cancers and transformed cell lines making it an attractive marker for tumor formation. The objective of this study was to survey CALML3 localization in normal epidermis and in hyperproliferative skin diseases including actinic keratosis, squamous and basal cell carcinoma as well as verruca and psoriasis and to compare CALML3 immunoreactivity with the proliferation marker Ki-67.MethodsParaffin-embedded tissue sections from normal human skin and hyperproliferative skin disorders were examined by immunohistochemistry and analyzed for localization and expression of CALML3 and Ki-67.ResultsCALML3 was strongly expressed in differentiating layers of normal skin, staining the periphery in suprabasal cells and exhibiting nuclear localization in the stratum granulosum. CALML3 nuclear localization was inversely correlated to Ki-67 staining in each disease, indicating that CALML3 nuclear presence is related to terminal cell differentiation and postmitotic state.ConclusionsIncreased CALML3 expression in suprabasal layers is characteristic for differentiating keratinocytes in normal epidermis, and nuclear expression of CALML3 inversely correlates with expression of the proliferation marker Ki-67. This suggests that CALML3 is a useful marker for normal and benign hyperplastic epidermal development, whereas the loss of nuclear CALML3 indicates progression to a proliferative and potentially malignant phenotype.

Drebrin, an Actin-Binding, Cell-Type Characteristic Protein: Induction and Localization in Epithelial Skin Tumors and Cultured Keratinocytes

Isoform E2 of drebrin, an actin-binding protein originally identified in neuronal cells, has recently been identified in diverse non-neuronal cells, mostly in association with cell processes and intercellular junctions. Here, we report on the presence of drebrin in normal human skin, epithelial skin cancers, and cultured keratinocytes. Keratinocytes of normal epidermis contain almost no drebrin but the protein is readily seen in hair follicles. By immunohistochemistry and immunoblot, basal cell carcinomas (BCC) are rich in drebrin, and confocal laser scanning and immunoelectron microscopy show accumulation at adhering junctions, in co-localization with actin and partially with plaque proteins. In squamous cell carcinomas, keratoacanthomas, and in epidermal precancers, drebrin is heterogeneously distributed, appearing as mosaics. Primary keratinocyte cultures contain significant amounts of drebrin enriched at adhering junctions. When epithelium-derived cells devoid of drebrin are transfected with drebrin-enhanced green fluorescent protein, constructs accumulate in the cell periphery, and immunoprecipitation shows complexes with actin. During epidermal growth factor induced formation of cell processes, drebrin retains this junction association, as observed by live cell microscopy. Our results suggest novel functions of drebrin such as an involvement in cell-cell adhesion and tumorigenesis and a potential value in diagnosis of BCC.

Parathyroid hormone-related protein expression and secretion in a skin organotypic culture system.

Parathyroid hormone-related protein (PTHrP), an important factor in the pathogenesis of humoral hypercalcemia of malignancy, is produced by many normal tissues, including the epidermis, where it is thought to play a role in the regulation of keratinocyte growth and differentiation. Most in vitro studies of normal keratinocytes use monolayer cell cultures, which have limitations, including the inability to reproduce the stratified structure of the epidermis. The objective of this study was to investigate PTHrP production and secretion, and mRNA expression in skin organotypic cultures. The cultures consisted of an artificial dermis with differentiating keratinocytes grown at the air-liquid interface. Immunohistochemical assessment of cytokeratins 14 and 10/13, involucrin, and proliferative cell nuclear antigen (PCNA) demonstrated that keratinocytes differentiated in a manner similar to keratinocytes in normal epidermis. PTHrP expression was demonstrated in all viable layers of the epidermis, as well as in some fibroblasts of the collagen lattice by immunohistochemistry and in situ hybridization. Since most fibroblasts expressed alpha-smooth muscle actin, these cells were interpreted to be consistent with myofibroblasts. PTHrP expression by myofibroblasts suggests a possible role for PTHrP in the regulation of contractibility of these cells. PTHrP was also detected in conditioned media for 50 days. In conclusion, because of its superior tissue morphology and ability to induce organized keratinocyte differentiation, this culture system will be an excellent model to study the role of PTHrP in pathologic and physiologic processes involving the epidermis in vitro.

Expression of proliferating cell nuclear antigen in Spitz nevus

Background: Epithelioid and spindle cell nevus (ESN; Spitz nevus) is a histologically well-described entity. We hypothesized that the features of ESN may reflect activation by a proliferative stimulus. Objective: Nevocytes and keratinocytes in ESN and control specimens were examined for expression of the proliferation marker, proliferating cell nuclear antigen (PCNA). Methods: Standard immunohistochemical methods were used to examine the expression of PCNA in a series of ESN, other nevi, and malignant melanoma. Results: PCNA was detected in nevocytes in a significant percentage of ESN but not in other nevi. PCNA expression was increased in basilar keratinocytes in ESN when compared with staining of basilar keratinocytes in normal epidermis. In other melanocytic nevi and nonin-flamed melanoma, PCNA expression in keratinocytes was similar to that in normal control tissue. Conclusion: Increased PCNA labeling in nevocytes and keratinocytes in ESN suggests that a growth stimulus, present within some of these lesions, affects both keratinocytes and nevocytes.

Detection of cytokine-induced protein γ-immune protein-10 (γ-IP10) in atypical melanocytic proliferations

gamma-Immune protein-10 (gamma-IP10) is a cytokine whose expression has been shown to be induced by interferon-gamma. It is a member of a group of closely related cytokines (e.g., interleukin 8 and platelet factor 4) with chemotactic properties. gamma-IP10 has been detected in keratinocytes, lymphocytes, monocytes, and endothelial cells in immunologically mediated processes, such as positive tuberculin skin tests, and in growth-activated keratinocytes, such as in psoriasis. Keratinocytes in normal epidermis do not produce gamma-IP10. We tested the hypothesis that keratinocytes adjacent to dysplastic nevi and melanomas would produce gamma-IP10, perhaps as part of an immune response to a tumor, and that this response would not be seen in ordinary melanocytic nevi. We used an affinity-purified, polyclonal rabbit anti-gamma-IP10 antibody to examine 10 nevi with moderate to severe histologic dysplasia, one superficial spreading melanoma, and 10 compound melanocytic nevi with no features of dysplasia. As predicted, keratinocytes surrounding all of the cytologically atypical melanocytic lesions displayed strong staining with gamma-IP10. There was no staining of keratinocytes adjacent to ordinary melanocytic nevi. The observed keratinocyte staining with gamma-IP10 may be related to a host immune response to antigenically abnormal cells.

Transforming growth factor‐β1 localization in normal and psoriatic epidermal keratinocytes in situ

Transforming growth factor-beta 1 (TGF beta 1) is a potent inhibitor of epithelial cell proliferation and its effects on growth and differentiation have been extensively characterized in cultured keratinocytes. We used two TGF beta 1-specific polyclonal antibodies (anti-LC and anti-CC) to determine the presence of TGF beta 1 peptide in keratinocytes in sections of normal human skin in situ and in both plaque and nonplaque skin from individuals with psoriasis. In contrast to the differentiation phenotype expressed by keratinocytes in normal epidermis, keratinocytes in the psoriatic plaque exhibit a hyperproliferative/regenerative differentiation phenotype. Anti-TGF beta 1 staining was observed primarily in the epidermis. Anti-LC TGF beta 1 antibody stained nonproliferating, differentiated suprabasal keratinocytes intracellularly in normal skin but did not stain psoriatic plaques from five of seven patients. In contrast, anti-CC TGF beta 1 antibody stained suprabasal keratinocytes extracellularly in psoriatic plaques, but did not stain normal skin. Both anti-LC and anti-CC stained suprabasal keratinocytes intracellularly in nonplaque psoriatic skin. Thus, the conformation or structure of TGF beta 1 and its localization vary in keratinocytes with distinct differentiation phenotypes suggesting that TGF beta 1 is a potential modulator of keratinocyte differentiation in vivo. Selective association of TGF beta 1 with nonproliferating keratinocytes in the suprabasal layers of the epidermis and its exclusion from the proliferating keratinocytes in the basal layer suggest that it may be a physiological regulator of keratinocyte proliferation. In addition, the intracellular localization of TGF beta 1 peptide in both normal and psoriatic keratinocytes suggests that it is constitutively synthesized by epidermal keratinocytes in vivo.

Ultrastructural Localization of Lectin-Binding Sites in Normal Skin

The distribution of carbohydrate residues in keratinocytes of normal epidermis was studied. Normal skin was embedded in Lowicryl. Thin sections were incubated with concanavalin A (Con A), peanut agglutinin (PNA), wheat germ agglutinin (WGA), Ulex europaeus agglutinin I (UEA I), dolichos biflorus agglutinin (DBA), and soybean agglutinin (SBA). A positive reaction in the dermis, in the basal lamina (lamina densa, lamina lucida), intracellularly and within the plasma membrane including the desmosomes was obtained after incubation with Con A and WGA. PNA binding sites were found predominantly in the plasma membrane between the desmosomes. The labeling with Con A, WGA, and PNA was most pronounced in the upper stratum spinosum and granulosum. Incubation with UEA revealed heavy labeling of the keratohyalin granules and the cytoplasm of the corneocytes. Incubation with DBA and SBA revealed weak labeling of the keratinocytes. The study of the distribution of carbohydrate residues in normal epidermis is important, since alterations in this distribution might be linked to autoimmunity or malignant transformation.

Distribution Patterns of the OKM-5 Antigen in Normal and Diseased Human Epidermis

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Distribution patterns of the OKM 5 antigen in normal and diseased human epidermis

The distribution of OKM 5-positive dendritic cells in the epidermis was investigated in 75 cases of inflammatory dermatoses and in 14 cases of normal human skin by immunohistochemical and morphometric methods. Furthermore 6 cases of normal human skin, 14 cases of nevocellular nevi, 26 cases of malignant melanoma, 7 cases of contact dermatitis and 6 cases of mycosis fungoides have been examined with special emphasis on the expression of OKM 5 antigen on keratinocytes. OKM 5-positive dendritic cells were present in normal human epidermis at a density of 46 +/- 3.4 cells/mm2 section area. However, there was a significant increase in cutaneous drug eruptions (166 +/- 17.2 cells/mm2; U-test: p less than or equal to 0.05). Concerning OKM 5-positive keratinocytes, a mean percentage of 10.2% +/- 5.0% OKM 5-positive keratinocytes was found in nevocellular nevi, compared to 0.5% +/- 0.5% in the adjacent skin. The corresponding values for malignant melanomas were 52.5% +/- 3.4% (lesional epidermis) and 7.1% +/- 2.2% (adjacent epidermis). There were significant differences of both lesional and adjacent epidermis between nevi and melanomas (U-test: p less than or equal to 0.05). Our cases of contact dermatitis revealed a mean percentage of 19.3% +/- 6.7% OKM 5-positive keratinocytes, whereas in mycosis fungoides the corresponding value represents 42.7% +/- 6.2%. The differences between the percentage of OKM 5-positive keratinocytes in normal epidermis and contact dermatitis as well as mycosis fungoides were significant (U-test).(ABSTRACT TRUNCATED AT 250 WORDS)

Histologic Distribution of Staining by a Monoclonal Antibody (ψ-3) in Psoriasis and Occurrence of ψ-3 Antigen in Other Cutaneous Diseases

psi-3 is a monoclonal antibody that recognizes a 135,000 molecular weight structural component of maturing keratinocytes in psoriasis (the psi-3 antigen) but fails to bind to any constituent of keratinocytes in normal epidermis. This paper describes the occurrence of the psi-3 antigen in a variety of dermatopathologic conditions using immunoperoxidase (biotin-avidin-peroxidase) and immunofluorescence methods which show excellent concordance. In 35 of 36 specimens of psoriasis vulgaris, psi-3 antibody consistently immunolabels the cytoplasm of keratinocytes above the basal layer. At the edges of psoriatic plaques, psi-3 antibody staining extends for a variable distance into lesion-free epidermis. A similar pattern has been found in a certain number of other conditions described in the paper, including squamous cell carcinoma and condyloma acuminatum, but not Darier's disease, basal cell carcinoma, nor lamellar ichthyosis. In all but one condition, the outermost or basal layer of cells is never stained. The only disease in which the lowermost cell layer is stained is a lichen planus-like lesion. The occurrence of psi-3 antigen cannot be correlated with any histologic feature of psoriasis such as acanthosis, loss of the granular layer, or hyperproliferation. The antigen appears to be a unique keratinocyte constituent which is expressed in certain pathologic conditions and which is not detected by any other histologic or immunophenotyping method. It is a potentially valuable addition to the panel of antibodies available for characterizing epithelial cells.