Malignant Skin Diseases Research Articles

TGF-α Is Widely Expressed in Differentiated as well as Hyperproliferative Skin Epithelium

Transforming growth factor-alpha (TGF-alpha) is a potent mitogen for epithelial cells that is expressed at low levels in normal epidermis and overexpressed in psoriasis. Epidermal growth factor (EGF) has been shown to inhibit hair growth but stimulate the growth of sebaceous and sweat glands, suggesting a potential role for a member of the EGF/TGF-alpha family in the normal development and function of skin appendages as well as epidermis. The present work demonstrates TGF-alpha protein in eccrine ducts, and eccrine, sebaceous, and apocrine glands. The proliferative dermal hair bulb does not express TGF-alpha in contrast to the differentiated outer root sheath hair follicle epithelia. In addition, hyperproliferative skin diseases including bullous congenital ichthyosiform erythroderma, squamous cell carcinoma, and psoriasis show increased TGF-alpha expression. Thus, TGF-alpha may play a role in the morphogenesis and function of normal skin appendages and its overexpression is common in benign and malignant hyperproliferative skin diseases.

The distribution of immuno-reactive tenascin in the epithelial-mesenchymal junctional areas of benign and malignant squamous epithelia

Tenascin is an extra cellular matrix glycoprotein which is distributed in the mesenchyme surrounding various organs during embryogenesis. It has also been demonstrated in some normal adult tissues and in the matrix of human tumours. The present study has been carried out to analyse the distribution of tenascin in non malignant and malignant skin disorders, in squamous cell carcinomas of the head and neck, in squamous cell carcinoma xenografts and in a squamous cell carcinoma cell line grown on collagen gel. Immunohistochemical localisation of tenascin was performed, using a monoclonal antibody specific for tenascin, by the indirect immunoperoxidase method with silver enhancement. Tenascin was heterogeneously distributed in the extra cellular matrix of squamous cell carcinomas and in squamous cell carcinoma xenografts. It was absent in basal cell carcinoma and in the squamous cell carcinoma cell line grown on collagen gel. The distribution of tenascin in squamous cell carcinoma and basal cell carcinoma is discussed in relation to tumour invasion and differentiation.

The significance of Leu 8 negative T cells in lymphoid skin infiltrates: malignant transformation, selective homing or T-cell activation?

The expression of Leu 8 was studied on skin biopsies from a large group of patients with benign and malignant skin disorders and correlated with the expression of T-cell differentiation antigens and activation markers. The effect of in vitro stimulation of peripheral blood T cells and T-cell subsets on the expression of Leu 8 antigen was also determined. In all the skin diseases studied an inverse relationship was found between the proportions of cells expressing Leu 8 and HLA-DR. A deficiency of Leu 8 positive cells was not specific for mycosis fungoides, but was also found in several reactive dermatoses. Stimulation of peripheral blood cells with phytohaemagglutinin (PHA), concanavalin A (Con A), and anti-CD3-PMA resulted in a considerable decrease of Leu 8 antigen expression on day 3 in both CD4+ and CD8+ T cells. These data suggest that the low proportion of Leu 8+ T cells in mycosis fungoides and several reactive skin disorders is not related to malignant transformation or selective homing of Leu 8- T cells, but probably results from local T-cell activation.

Keratinocyte and epidermal leukocyte expression of CD36 (OKM5) in benign and malignant skin diseases

CD36 recognizes a 88 Kd glycoprotein, found on different cell populations involved in immunoregulation and are induced on keratinocytes by in vitro treatment with gamma-interferon. Therefore, we obtained skin biopsies from 48 patients with various dermatological diseases and from 5 healthy volunteers and stained these with monoclonal antibodies OKM5 (CD36), anti-HLA-DR and OKT6 (CD1a) using a three stage immunoperoxidase method. In normal skin, CD36 expression was not observed. In contrast, keratinocytes in diseased skin were CD36+. In most cases, the staining was restricted to the stratum granulosum and the stratum spinosum, but in psoriasis, squamous cell carcinoma and lymphomatoid papulosis, more extensive staining of keratinocytes was seen. In addition, CD36+ epidermal leukocytes were found in allergic patch-test infiltrates and in mycosis fungoides. The findings of CD36 expression by epidermal cells within a broad spectrum of dermatological diseases indicate a role for these cells in the regulation of immune reactions in the skin.

Synthetic Retinoids Inhibit the Antigen Presenting Properties of Epidermal Cells in Vitro

The clinical efficacy of retinoids in benign and malignant skin diseases involving immune mechanisms suggests that they affect the immunologic functions of the epidermis. However, these effects have yet to be demonstrated. The action of vitamin A (retinol) and the synthetic retinoids, isotretinoin, etretinate, acitretin, and arotinoid-free acid have been studied on the lymphocyte proliferation induced by phytohemagglutinin (PHA), by the mixed lymphocyte reaction (MLR), and the mixed epidermal cell-lymphocyte reaction (MECLR). The results for PHA-induced proliferations were highly variable for all the retinoids. However, in MECLR, the synthetic retinoids consistently reduced the proliferation by 20%-30%. This occurred at therapeutic drug concentrations of about 10(-7)M. In MLR, a minor decrease of 10%-15% was only found for higher concentrations (10(-5)M). Retinol induced no effect in either reaction. Further analysis of acitretin on MECLR showed that it reduced lymphocyte proliferation in a dose-dependent fashion. This reduction was combined with a decrease in cytotoxic T-lymphocyte induction (CTL). Addition of 10(-6)M acitretin at various times also revealed that its presence at cell culture initiation was necessary to inhibit proliferation significantly. Furthermore, cell treatments prior to MECLR showed that exposure of epidermal cells to acitretin was essential to produce this inhibition, suggesting that it acts directly on epidermal cells. Consequently, it is suggested that the specific inhibitory effect of synthetic retinoids on lymphocyte activation in MECLR may partly account for their therapeutic action on the skin.

Altered Keratin Expression in Benign and Malignant Skin Diseases Revealed with Monoclonal Antibodies

We studied keratin expression in benign epidermal skin diseases, and in Bowen's disease by using three monoclonal cytokeratin antibodies. In adult normal skin, these antibodies bind only to the follicular epithelium (PKK1), the basal keratinocytes (PKK2), or the suprabasal cells in interfollicular epidermis (KA5). Additionally, in fetal epidermis, the PKK1 antibody reacts with basal keratinocytes. In psoriasis and lichen planus, the PKK2 antibody distinctly revealed all epidermal cell layers by immunostaining. However, a negative basal cell-like layer was revealed in both lesions with the KA5 antibody. In pityriasis rubra pilaris, the basal cell layer was uniformly stained with the PKK2 antibody, but only some keratinocytes in upper cell layers showed fluorescence and, in chronic eczema, the 3-4 lowest epidermal cell layers were reactive. The PKK1 antibody did not stain interfollicular keratinocytes in any of the benign proliferative skin diseases studied. In Bowen's disease, a heterogeneous staining pattern with varying intensity among individual cells was seen with all of the antibodies used. Our results suggest different changes in keratin expression in chronic benign and malignant epidermal diseases that may reflect the mechanisms behind these changes.

Detection of transforming growth factor alpha in normal, malignant, and hyperproliferative human keratinocytes.

Transforming growth factor alpha (TGF-alpha) is a 50-amino acid peptide, previously demonstrated only in transformed cell lines and human tumors, which is structurally homologous to epidermal growth factor (EGF). TGF-alpha expression in keratinocytes from normal individuals, patients with psoriasis, and patients with malignant skin diseases was investigated using an mAb raised against synthetic human TGF-alpha. mAb A1.5 reacted with TGF-alpha, but not EGF, in a sensitive ELISA. Keratinocytes in eight nodular basal cell carcinomas, one morpheic basal cell carcinoma, and one squamous cell carcinoma demonstrated intense membranous immunoperoxidase staining with mAb A1.5. Of even greater interest was the observation that the overlying normal epidermis, as well as the epidermis from five normal skin specimens, were stained by the mAb. Keratinocytes in plaques from 18 psoriasis patients were more intensely stained than those from normal skin. Cultured normal keratinocytes demonstrated membranous staining with mAb A1.5. Absorption of mAb A1.5 with synthetic human TGF-alpha completely removed the reactivity of mAb A1.5 with both basal cell tumors and normal epidermis. The demonstration of TGF-alpha in normal keratinocytes suggests that it plays a role in normal keratinocyte growth, wound healing, and in the pathogenesis of acanthosis.

Mononuclear cell patterns in the skin. An immunohistological and morphometrical analysis.

A large variety of skin diseases is characterized by the presence of mononuclear cell infiltrates in the dermis and to some extent in the epidermis. We have investigated frozen material of 566 lesions of benign and malignant skin diseases by immunohistological methods with a panel of 20 monoclonal antibodies; quantitative studies using computer-assisted image analysis were additionally performed in 80 specimens. In all reactive lesions, mononuclear cells were arranged in distinct compartments simulating the architecture of normal lymphatic tissue. A qualitatively uniform T cell compartment (mainly helper-inducer T lymphocytes, suppressor-cytotoxic T lymphocytes, Langerhans' cells) with slight quantitative differences was found in all inflammatory skin diseases, in peritumoral infiltrates, and also in normal skin. The B cell compartment (B lymphocytes, few T lymphocytes and monocytes) is only rarely seen (some B cell lymphomas and pseudolymphomas). The monocyte compartment (monocytes, few T lymphocytes) associated with a T cell compartment is typical for granulomatous skin lesions. The epidermis forms a separate functional region, which might evolve into a lymphoepithelial compartment in diseased skin. Low-grade malignant lymphomas of the skin display similar architectural patterns as reactive lesions, whereas high-grade malignant lymphomas do not. Obviously the mononuclear cells in the skin are usually arranged in compartments and form a limited number of distinct patterns. As similar patterns are found in diseases that are completely different with respect to clinical appearance, histological features, cause, and outcome, the patterns are not disease specific but reflect general anatomical-functional relationships of inflammatory cells and the skin.

Immunohistological aspects of cutaneous T-cell infiltrates.

SUMMARY In many benign and malignant skin disorders, lymphocytes and associated monocytic cells predominate among the various types of cells infiltrating the dermis and/or the epidermis. Immunohistological techniques utilized for the purpose of combining immunological parameters and morphological criteria have been reviewed. The rosetting techniques, enzyme histochemistry and immunohistological techniques using polyclonal and monoclonal antibodies were reported for light microscopy. In the field of immunoelectron microscopy, immunoperoxidase labelling and the immunogold technique were also subject to discussion. These techniques may be supplemented by various methods of cell biology (obtaining enriched cell suspensions of infiltrate cell subsets). The possiblity of quantitative studies was considered. Results obtained in cutaneous pathology using these techniques were reviewed and discussed. Among others, human skin tuberculin lesions and DNCB skin tests represent models of delayed hypersensitivity. Infiltrates in the following conditions were presented: non-tumoral lesions (lichen planus, erythema multiform, erythema fixum, GVH and allograft rejection, cutaneous sarcoidosis and other infiammatory infiltrates); in lymphomas and related conditions; in tumoral lesions (warts and papillomas, peritumoral infiltrates around epidermal neoplasias). Finally Langerhans cells in infiammatory dermatosis, in skin tumours and in histiocytosis X were considered.

Plastic and Reconstructive Treatment of Facial Skin Lesions

There are two reasons why I chose skin defects for my lecture today: on the one hand, we are very often confronted with malignant skin diseases in the head-neck region, especially in the older patient. On the other hand, we are convinced that the head and neck surgeon is especially qualified to treat this kind of disease as he considers not only the surface of the disease but also underlying structures and tissues like salivary glands, nerves, bone, and the pneumatic system of the sinuses. I shall try now to deal with the distinct regions of the face separately wherever possible. Extensive soft tissue defects of the frontal region may be reconstructed using a bridge flap from the scalp or with full-thickness skin grafts taken from the supraclavicular fossa or from the groin. The advantage of constructing a bridge flap is that it has the same thickness as the original tissue. However, this flap is usually used only as provisional cover, since hair growth is undesirable in the forehead area. The free skin graft is in many cases more advantageous, as large areas of it can be provided without causing large-scale defects in the donor region. In the frontal region, however, free skin grafts should be used only after an interval of several weeks, when the level of the defect has been elevated by granulation tissue. This superficial layer of granulation tissue must be removed immediately prior to skin graft transplantation. The second granulation tissue layer has a more substantial structure and provides excellent nourishment for free skin grafts. Frontal skin defects originating from frontal skin flap excision are frequently closed by a procedure known as the H-shape plastic. This plastic is capable of closing defects up to a maximum width of 3 cm. An incision close to the eyebrows is frequently not required. In such cases, the plastic resembles a T-shape instead of an H-shape. As the scars in the donor region are usually dissolved by the Z-plasty technique, they are almost invisible in many cases, for example after raising a frontal skin flap for reconstructing defects in the nasal region.

The Sézary cell in the blood of patients with mycosis fungoides.

A study of 96 patients with an original diagnosis of mycosis fungoides revealed that 84 did not have circulating Sézary cells. Of the 12 patients with Sézary cells, only 3 had more than 15%. The 12 patients had benign and malignant skin diseases. The Sézary cell does not seem to have any importance in the pathogenesis of mycosis fungoides.

ANOTHER LOOK AT KERATOACANTHOMA

In a retrospective survey over a three-year period there were found 199 patients with keratoacanthoma among 3,180 patients with malignant skin disease. The clinical features of the series are presented, with particular attention to the diagnostic error. Amongst experienced clinicians there was an 8.5% error in calling an invasive squamous cell lesion a keratoacanthoma. Pertinent features of the clinical nature of keratoacanthoma with particular reference to immunological surveillance are discussed.

Antibody dependent cell mediated cytotoxicity (ADCC) in malignant and benign skin disease

Antibody dependent cell mediated cytotoxicity (ADCC) in malignant and benign skin disease

Ultrastructure of Abnormal Cells

A cell with characteristic ultrastructural features was found in tissues from all six Sezary patients studied. This cell has a highly irregular, serpentine nucleus, which in three-dimensional reconstruction appears to be cerebriform and composed of highly folded sheets. Abnormal cells were also found in tissues from 34 of 38 patients with mycosis fungoides (MF). The nuclei of these MF cells were also irregular, but less so than Sezary cell nuclei. These MF cells were also found in six of eight patients with parapsoriasis en plaque. In order to determine the diagnostic specificity of these abnormal MF cells, skin from 33 control patients with a variety of malignant and benign skin diseases was studied. Cells similar to MF cells were found in skin from three of these control patients; so, it appears that the presence of this unusual cell is not absolutely specific for the diagnosis of MF.

Ultrastructure of abnormal cells in Sezary syndrome, mycosis fungoides, and parapsoriasis en plaque.

A cell with characteristic ultrastructural features was found in tissues from all six Sezary patients studied. This cell has a highly irregular, serpentine nucleus, which in three-dimensional reconstruction appears to be cerebriform and composed of highly folded sheets. Abnormal cells were also found in tissues from 34 of 38 patients with mycosis fungoides (MF). The nuclei of these MF cells were also irregular, but less so than Sezary cell nuclei. These MF cells were also found in six of eight patients with parapsoriasis en plaque. In order to determine the diagnostic specificity of these abnormal MF cells, skin from 33 control patients with a variety of malignant and benign skin diseases was studied. Cells similar to MF cells were found in skin from three of these control patients; so, it appears that the presence of this unusual cell is not absolutely specific for the diagnosis of MF.