S. Husz

Effects of the neuropeptides substance P, calcitonin gene-related peptide, vasoactive intestinal polypeptide and galanin on the production of nerve growth factor and inflammatory cytokines in cultured human keratinocytes

Neuropeptides released from the cutaneous sensory nerve endings have neurotransmitter and immunoregulatory roles; they exert mitogenic actions and can influence the functions of different cell types in the skin. The aims of this study were a systematic investigation of the effects of the neuropeptides substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and galanin (GAL) on the inflammatory cytokine production (IL-1alpha, IL-8 and TNF-alpha) of the keratinocytes, and a study of their role in the production and secretion of nerve growth factor (NGF) and its precursor molecule (proNGF). Cultures of normal human keratinocytes were treated with 10(-8)M SP, CGRP, VIP or GAL for 30 min. After different time intervals, cells were harvested for total RNA isolation; in addition, cell lysates and supernatants were collected. The effects of the neuropeptides on the mRNA expressions of the different cytokines and NGF were investigated by Q-RT-PCR and the protein levels were studied by means of ELISA assays and Western blotting. Each of the four neuropeptides induced increases in the expressions of IL-1alpha, IL-8 and TNF-alpha mRNA. Increases appeared in the amount of the IL-1alpha protein in the supernatants of neuropeptide-treated cells, and the IL-8 secretion was mildly elevated, while secretion of TNF-alpha remained undetectable. The four neuropeptides increased the NGF mRNA expression to different extents. In the cell lysates of the keratinocytes, only proNGF could be detected, its concentration in the neuropeptide-treated cells being approximately twice that in the time-matched controls. Both control cultures and neuropeptide-treated cultures were found to secrete proNGF and mature NGF, but neuropeptide-treated cell cultures produced markedly higher (3-7-fold) amounts of NGF-like immunoreactive materials. The results demonstrated that neuropeptides released from cutaneous nerves after an injurious stimulus are able to induce an upregulation of IL-1alpha and IL-8 production; they are additionally able to influence the expressions of proNGF/NGF and their secretion from the keratinocytes. These findings may contribute toward an understanding of the neural influence on skin health and disease.

Immunoglobulin‐ and complement‐bearing lymphocytes in allergic contact dermatitis and atopic dermatitis (eczema)*

The distribution pattern of immunoglobulin‐ or complement‐bearing lymphocytes in healthy controls and in patients with contact dermatitis and with atopic dermatitis has been investigated. A significant predominance of IgD‐ and C3‐bearing lymphocytes was found in ten patients with contact dermatitis. In atopic dermatitis patients the number of IgE‐ and to a lesser extent of C3‐bearing lymphocytes was increased. The results indicate induction of stimulation of two distinct classes of immuno‐logically competent lymphoid cell lines, one in contact and the other in atopic dermatitis. The increased number of B‐cells in dermatitis indicates a decreased T‐cell population and suggests a deficiency or disturbance of the recirculation of this population.

Coexistent atypical bullous pemphigoid and systemic lupus erythematosus

A vesiculobullous eruption with clinical and histological features of bullous pemphigoid developed in a 38‐year‐old woman with proved systemic lupus erythematosus. The patient had a sulphone responsive blistering disease that was characterized by pruritic subepidermal bullae and linear, predominantly IgA basement membrane zone deposition and IgA pemphigoid antibodies in her sera. Because both diseases are associated with immune complexes of special immunoglobulin classes, this association may not be entirely fortuitous.

Multicentre study of fragrance allergy in Hungary. Immediate and late type reactions.

The authors followed the frequency of fragrance contact sensitization in Hungary in a multicentre study in the years 1998 and 1999. A total of 3604 patients were tested with fragrance mix (FM), and positive reactions were observed in 294 (8.2%). In 160 FM hypersensitive patients, the study was continued with patch testing of the mix constituents (cinnamic alcohol, cinnamic aldehyde, eugenol, amyl cinnamic aldehyde, hydroxycitronellal, geraniol, isoeugenol, oak moss absolute). Of the patients tested, 70.6% produced positive reactions to the constituents. FM contact sensitization was mainly observed in female patients (74.4%). The incidence of contact urticaria in FM hypersensitive patients was 6.1%. Simultaneous patch test trials of other environmental contact allergens, in both early and late evaluations, mainly confirmed hypersensitivity reactions to balsams. Female dominance of hypersensitivity reactions observed during testing the individual components of the mix was striking (82.4%). In positive skin reactions, cinnamic alcohol, isoeugenol and oak moss provoked skin symptoms most frequently. We also tested the 104 patients who produced negative reactions to FM with the constituent individual allergens, with 11.9% positive incidence. The clinical symptoms of the patients were above all manifest in the form of contact eczema, located on the hands, face, eyelids and axillae. With this study, the authors, members of the Hungarian Contact Dermatitis Research Group, call attention to one of the most frequent allergens in the environment.

Antineutrophil cytoplasmic antibodies in patients with systemic lupus erythematosus

Summary  We determined the prevalence of antineutrophil cytoplasmic antibodies (ANCAs) in patients with systemic lupus erythematosus (SLE) and evaluated the correlation between ANCA positivity and clinical features. Forty‐one patients with SLE and two control groups were examined. One of the control groups consisted of 15 patients with systemic vasculitis, and the other of 12 healthy blood donors. A quantitative enzyme‐linked immunosorbent assay technique was used to measure the serum cytoplasmic ANCA (cANCA) and perinuclear ANCA (pANCA) levels. cANCA positivity was found in three patient samples, and pANCA positivity in 10 SLE patients. The occurrence and titres of both ANCA types in SLE patients were similar to those in healthy controls and significantly lower than those in patients with systemic vasculitis. The clinical picture and antibody profile were similar in ANCA‐positive and ANCA‐negative SLE patients. We conclude that measurement of ANCAs does not provide any additional diagnostic or prognostic data in SLE.

CYCLOSPORIN FOR DYSTROPHIC EPIDERMOLYSIS BULLOSA

A 65-year-old woman with epidermolysis bullosa since childhood was first admitted to our clinic in September, 1987. At first we treated the patient with high doses of vitamin E, phenytoin, prednisolone, and erythromycin but without sucess. When we read a report of the use of cyclosporin in a patient with epidermolysis bullosa acquisita, we began treatment with the above drug combination plus cyclosporin 1 mg/kg daily. Soon after the interruption of cyclosporin therapy, the blisters and erosions reappeared again. Cyclosporin was reintroduced and significant improvement was noted one month later

Galanin receptor expression in cultured human keratinocytes and in normal human skin

Abstract  Galanin (GAL) is a biologically active neuropeptide that is widely distributed in the nervous system. GAL exerts diverse action via the GAL receptors (GALR1, GALR2, and GALR3), which belong in the superfamily of G‐protein‐coupled transmembrane receptors. In human skin, GAL‐like immunoreactivity has been reported in free nerve endings and fibers of the dermis. The extraneuronal expression of GAL has also been demonstrated. Although the GALRs are essential for biological functions, the expressions of different GALR subtypes in cultured human keratinocytes have not yet been investigated. The aim of our study was to investigate the mRNA and protein expressions of the different GALRs in the HaCaT immortalized keratinocyte cell line and in cultured human keratinocytes. When reverse transcription (RT)‐polymerase chain reaction (PCR) was used with different GALR‐specific primers, only GALR2 mRNA was identified in cultured HaCaT cells and keratinocytes. Sequencing of the PCR products proved the presence of GALR2 mRNA in the keratinocytes. The presence of GALR2 protein was next investigated, using a polyclonal antibody against human GALR2. Both the HaCaT cells and the cultured keratinocytes displayed specific immunohistochemical staining, with higher intensity on the surface of the keratinocytes. Immunohistochemical investigations of normal human skin specimens revealed that GALR2 was expressed with high intensity in the basal layer of the epidermis and also around the hair follicles in the dermis. GAL treatment of the keratinocytes resulted in an increase in cytosolic Ca2+ concentration, suggesting that GALR2 is a functional receptor. Further studies are necessary to clarify the biological effects of GAL in the skin.

Impairment of axon reflex vasodilatation after herpes zoster

Examination of the neurogenic inflammatory response of the skin renders direct information on the functional state of the chemosensitive sensory nerve endings involved in neurogenic vasodilatation of skin vessels. Using a recently developed diagnostic test, the present study furnishes experimental evidence on the impairment of axon reflex vasodilatation following herpes zoster. The possible mechanisms by which herpes zoster may effect the vascular reactions of the skin are briefly discussed.

Juvenile pemphigus foliaceus

A 7‐year‐old girl with generalized erythematous, scaling plaques and vesiculobullous lesions on the extremities was diagnosed as having pemphigus foliaceus. Lesional direct immunofluorescence revealed intercellular IgG, IgA and C3 deposition. The patients serum gave positive reactions against one epitope of desmoglein 3 and the epitope of desmoglein 1 in enzyme‐linked immunosorbent assays, but the blood sample for indirect immunofluorescence did not display any circulating antibodies. The patient was successfully treated systemically with prednisolone and dapsone. Currently, she is taking dapsone, 12.5 mg daily. She has been free of lesions for the last 3 years.

Drug-induced erythema multiforme-like bullous pemphigoid.

Sir, Bullous pemphigoid (BP) is a common autoimmune blistering disease in the elderly, with an early manifestation of urticarialike and pruritic erythematous lesions which subsequently lead to the formation of bullae. The diagnosis is con¢rmed by the histological and immuno£uorescence ¢ndings (1). Despite its autoimmune origin, some reports demonstrate a clear induction of BP by PUVA and drugs (2). We report here a case in which BP was induced by antipsychotics and antidepressant drugs. During the recovery period, doxycycline was observed to exert a positive e¡ect on the BP.