Immunohistochemical demonstration of migration inhibitory factor in different types of urticaria

Because urticarial lesions can persist for extended periods of time, we have investigated the histochemical expression of an antibody against the cytokine macrophage inhibitory factor in 23 patients with different types of urticaria. Positive staining of upper and middermal dendritic cells was noted in sections from all three biopsy specimens of acute urticaria, eight of chronic urticaria, and all six of urticaria pigmentosa lesions. In all but one biopsy specimen, endothelial cells reacted as well. In three sections (two chronic urticaria, one urticaria pigmentosa), luminal lining cells of sweat glands were also noted to stain positively. In contrast, lesional skin from all eight patients with pressure urticaria was negative, as was the clinically normal skin of all patients, with the exception of one patient with urticaria pigmentosa. The data suggest that cytokines may be involved in lesions of acute type immunologic processes and that they need not be expressed in delayed type reactions.     

Expression of Monocyte/Macrophage Markers (CD13, CD14, CD68) on Human Keratinocytes in Healthy and Diseased Skin

The results of several investigations prove that, in special circumstances, human keratinocytes (HKs) synthesize and express cell surface moieties characteristic of effector and/or accessory cells of the immune system, such as CD16, CD36, HLA-DR, and intercellular adhesion molecule-1 (CD54), which are all detectable on the surfaces of macrophages. In the present study, skin biopsies from healthy volunteers, from positive tuberculin skin tests, and from patients with acute urticaria (AU), lichen planus (LP), psoriasis vulgaris (PV), mycosis fungoides (MF), and purpura pigmentosa chronica (PPC) were investigated by means of a multistep immunoperoxidase method to examine the reactivity of the HKs with a panel of monoclonal antibodies (MABs) characteristic of monocyte/macrophage cell lines. In biopsies obtained from positive tuberculin tests and from clinically involved skin of patients with LP, PV, MF, or PPC, a multifocal, positive peroxidase reaction was observed on the membranes of HKs of the basal and suprabasal cell layers when the MABs OKM13 (CD13), OKM14 (CD14), and Dako-Macrophage (CD68) were used. In contrast, specific staining of the HKs was not observed with the same antibodies in the biopsies of healthy volunteers or of patients with AU or in the uninvolved skin specimens obtained from the other patients. The HKs of PV, LP, MF, PPC, and AU patients and those of the healthy subjects all failed to give positive reactions when MABs against CD11b, CD15, or CD33 were used. The published data supplement the known surface characteristics of HKs, reflecting their stage of activation and differentiation.     

Expression of complement receptor CR2 (CD21) on human subcorneal keratinocytes in normal and diseased skin.

Human keratinocytes are able to synthesize and express cell surface moieties characteristic of effector and/or accessory cells of the immune system (CD16, CD36, HLA-DR, intercellular adhesion molecule-1). In the present study, skin biopsies from healthy volunteers, from patients with psoriasis vulgaris (PV), mycosis fungoides (MF), purpura pigmentosa chronica (PPC), acute urticaria (AU) and from positive tuberculin skin tests were investigated with regard to the reactivity with the monoclonal antibodies to complement receptors CR1 CR2 and CR3 by means of a multistep immunoperoxidase method. In the clinically involved skin of all patients with PV, MF or PPC, and in biopsies obtained from positive tuberculin tests, specific epidermal intercellular staining with OKB7 and Leu anti-CR2 was seen on subcorneal keratinocytes. This finding suggests a differentiation-linked expression of CR2 on human keratinocytes in cytokine-mediated skin diseases whereas CR1 and CR3 are apparently not expressed.     

Persistent spontaneous erythema caused by topical use of fumaric acid monoethyl ester--an obligate mast cell degranulation?

In 12 healthy volunteers the topical application of monoethyl fumarate caused a spontaneous persistent erythema. Systemic application induced flush. In urticaria pigmentosa the influence was significant higher than in normal skin. This fact suggests a mast cell degranulation caused by monoethyl fumarate.     

Expression of stem cell factor in cutaneous mastocytosis

Stem cell factor has recently been identified as a potent growth factor for bone marrow stem cells, melanocytes and mast cells. In order to evaluate its possible role in human mastocytosis, skin lesions from 13 patients with urticaria pigmentosa and five patients with mastocytomas, and normal skin specimens from five healthy donors were studied by immunohistochemistry, using polyclonal and monoclonal (hkl-12) antibodies against stem cell factor, and a monoclonal antibody (YB5.B8) against its receptor, the c-kit proto-oncogene product. Stem cell factor expression was noted in all sections studied, with an equal distribution pattern for both antibodies, but a weaker intensity with the hkl-12 reagent. Cytoplasmic staining was noted in keratinocytes, Langerhans cells, sweat gland ductal lining cells, mast cells, endothelial cells and spindle-shaped dermal stromal cells. An intense, diffusely granular reaction pattern was noted in all cells, except for a sparse, coarsely granular pattern in mast cells and stromal cells. In urticaria pigmentosa, staining was weaker in keratinocytes, but more prominent in Langerhans cells. In all sections, toluidine blue-positive mast cells and TA 99-positive basal epidermal melanocytes were the only cells to react with the c-kit antibody. Mastocytomas and urticaria pigmentosa lesions thus exhibit different patterns of stem cell factor expression. However, a possible pathogenetic role of this factor in mastocytosis remains to be determined.     

Macrophage Markers 25F9 and 27E10 on Human Keratinocytes in Normal and Diseased Skin

In the present study, an immunohistochemical analysis was made of the expression pattern of different macrophage markers such as 25F9, 27E10 and RM 3/1 on the surface of human keratinocytes (HK) in biopsies obtained from healthy volunteers and from patients with lichen planus, chronic cutaneous graft-versus-host reaction, mycosis fungoides, and purpura pigmentosa chronica. In biopsies from the healthy volunteers and from both clinically involved and uninvolved skin of the patients, the HK of the basal cell layer exhibited a specific peroxidase-positive reaction when the monoclonal antibody against 25F9 was used. Lesional HK from all patients studied displayed 25F9 and 27E10 in nearly the entire epidermis. The present findings provide further evidence that HK and macrophages share a number of common cell surface moieties.     

Increased tryptase levels in suction-blister fluid from patients with urticaria

The levels of tryptase in the suction-blister fluid from patients with chronic urticaria, urticaria pigmentosa, cholinergic urticaria, urticarial dermographism, prurigo of unknown origin, eczema, psoriasis, atopic dermatitis, and from healthy controls were studied. The blister fluid from controls contained up to 15 micrograms/l of tryptase, whereas that from patients with active urticaria contained greater than 50 micrograms/l. This study demonstrates that patients with urticaria have mast cells that readily release tryptase in both the lesional and non-lesional areas of skin.     

Expression of beta-2 integrin molecules on human keratinocytes in cytokine-mediated skin diseases.

Integrins are cell surface molecules of importance in a wide variety of cellular functions, including morphogenesis, cell migration and cell matrix interactions. The beta-2 (B2) integrin (leukocyte integrin, CD11/CD18) subfamily comprising three members, each consisting of a shared beta subunit (CD18) non-covalently associated with unique alpha subunits (CD11a, CD11b, CD11c). In the present study, we have analysed the expression pattern of B2 integrins on the surface of human keratinocytes (HKs) in biopsies obtained from healthy volunteers, from positive tuberculin skin tests and from patients with acute urticaria (AU), lichen planus (LP), psoriasis vulgaris (PV), mycosis fungoides (MF) or purpura pigmentosa chronica (PPC). In biopsies obtained from positive tuberculin tests and from the clinically involved skin of patients with LP, PV, MF or PPC, a multifocally occurring, suprabasal peroxidase-positive reaction was observed on the membranes of the HKs when the monoclonal antibodies (MABs) Dako CD11a, Dako-p150, 95 or Dako CD18 were used. In contrast, no specific staining of the HKs was observed with the same MABs in biopsies from healthy volunteers, from patients with AU and in the uninvolved skin specimens obtained from the other patients. The HKs from PV, LP, MF, PPC and AU patients and those from the healthy subjects failed to give a positive reaction when the MAB against CD11b (OKM1) was used. Our present findings provide further evidence that HKs may be actively involved in cell adhesion processes.     

Ultrastructural findings in tissue mast cells in urticaria pigmentosa

In a study on 10 patients suffering from urticaria pigmentosa, biopsies of skin lesions were electron microscopically investigated. In contrast to normal skin, the mast cells showed irregular, partly bilobed nuclei, prominent nucleoli, hypogranulation, and rather small granules; giant granules were rarely observed. In addition, we frequently found degranulation, and the development of microvilli was much more prominent than with normal mast cells. These deviations from normal skin represent at least in part functional alterations. However, some of the findings suggest that urticaria pigmentosa may be considered a neoplastic disease of the tissue mast cells, although clear evidence is still lacking. In any case, our findings show that mast cells in mastocytosis are not identical with those in normal skin.     

Two novel mast cell phenotypic markers, monoclonal antibodies Ki-MC1 and Ki-M1P, identify distinct mast cell subtypes

Summary In order to identify more specific or selective mast cell markers, the reactivity of two monoclonal antibodies, Ki-MC1 and Ki-M1P, was studied by immunohistochemistry in two human cell lines (mast cell line HMC-1, basophilic cell line KU812), in mast cells cultured from blood precursors, in adherent mononuclear cells from peripheral blood, and in mast cells of tissue sections from 1 3 urticaria pigmentosa lesions, live mastocytomas and live normal skin specimens. Toluidine blue staining, fluorescence staining with FITC-conjugated avidin, and immunohistochemical staining (APAAP) with other mast cell reactive monoclonal antibodies, was performed for comparison. Double staining with the APAAP method, using the Ki-antibodies and toluidine blue, was also carried out. Both Ki-antibodies showed reactivity for skin mast cells, but with a different staining pattern. In addition, the Ki-MC1 antibody did not react with the cell lines, and reacted only with a few peripheral blood mononuclear cells and cultured mast cells. In contrast, the Ki-M1P antibody reacted with almost all cultured mast cells and blood mononuclear cells, but stained only about one-half of lesional and one-fifth of normal skin mast cells. Ki-M1P also reacted with many toluidine blue-negative dermal cells, particularly in urticaria pigmentosa. Ki-MC1 antibody can thus be considered as a useful additional marker for normal skin mast cells. In contrast, the Ki-M1 P antibody primarily identifies immature mast cells and monocytes/macrophages, suggesting that these cell types probably originate from the same bone marrow precursor.     

自身免疫性甲状腺疾病与慢性荨麻疹关系探讨

目的：探讨慢性荨麻疹与自身免疫性甲状腺疾病之间的关系。方法：采用放射免疫法对58例慢性荨麻疹患者及30例正常体检人群的甲状腺功能及抗甲状腺自身抗体进行了检测，结果：58例慢性荨麻疹患者中，有8例（13．8％）血清中存在抗甲状腺球蛋白抗（TGA）；7例（12．1％）存在抗甲状腺微粒体抗体（TMA）；慢性荨麻疹组的TGA和TMA水平均显著高于对照组(t值分别为2．762和2．695，P＜0．01）。但两组的T4、RT3、FT3、TSH值比较差异均无显著性（P＞0．05）。结论：慢性荨麻疹的发病有自身免疫机制参与。     

Delayed pressure urticaria histologically resembles cutaneous late-phase reactions

In a recent study, all patients with delayed pressure urticaria (DPU) developed late cutaneous reaction (LCR) after intradermal injection of compound 48/80 and after skin testing with certain food antigens. In the present study, we analyzed the histologic changes in the pressure lesions and compared them with those found in normal skin injected with diluent and in LCR to 48/80. The study included five patients with DPU associated with chronic urticaria (CU) and four patients with CU but without DPU. Six to eight hours after pressure challenge and intradermal skin testing with 48/80 and diluent, skin biopsy specimens were obtained from the pressure lesions, the LCRs, and normal skin (diluent injection). Specimens were assessed by Giemsa staining of 1-micron sections and immunofluorescence of frozen sections. Total cells were counted in each specimen. Interstitial deposits of fibrin were observed by immunofluorescence in LCR and pressure lesions. The total numbers of infiltrating cells in the dermis among LCR sites and pressure lesions were not significantly different, while both LCR sites and pressure lesions contained significantly more infiltrating cells than did normal skin injected with saline diluent. The differential counts in LCR and DPU were mostly mononuclear cells. Infiltrates in the DPU and LCR were mostly perivascular. No histopathologic changes were seen at skin sites challenged with pressure in the control patients with CU without clinical manifestations of DPU. We conclude that lesions seen in DPU are morphologically similar to classic LCR.     

Treatment of urticaria pigmentosa using interferon alpha

Long-term treatment with interferon alpha (IFN-α) has recently been shown to reduce the bone marrow infiltrate and cutaneous lesions in systemic mast cell disease. We therefore administered this cytokine to six patients with urticaria pigmentosa for up to 12 months, using subcutaneous injections of 5×10⁶U, initially five times, and subsequently three times a week. The generally well-tolerated therapy resulted in marked improvement of the cutaneous symptoms, especially in three of the patients who suffered from very severe pruritus. Two of the patients with bone marrow infiltration showed normal findings after treatment. However, in none of the patients was there any change in the skin lesions, or decrease in the degree of cutaneous mast cell infiltration, as evidenced by light and electron microscopic examination. These findings indicate that IFN-α is highly effective in the control of symptoms, but otherwise does not influence the cutaneous lesions of urticaria pigmentosa.     

Photochemotherapy (PUVA) in the treatment of urticaria pigmentosa

Eight patients received PUVA for mastocytosis. Five women had typical adult-onset urticaria pigmentosa, without evidence of systemic disease. Another woman had suspected hepatic involvement while the remaining female had early-onset familial urticaria pigmentosa with morphologically atypical mast cells. The only male patient had cirrhosis with hepatic deposits of mast cells in addition to polycythaemia rubra vera. In all patients, except the man with systemic disease, there was reduced pruritus and wealing and partial to almost complete fading of the macules. The manifestations of urticaria pigmentosa recurred after treatment was discontinued. In both lesional and uninvolved skin there was no significant change in either the mean mast cell counts or mast cell ultrastructure after an average of twenty-seven PUVA exposures. In addition, PUVA did not cause a significant alteration in the histamine content of the skin. The beneficial effect of PUVA in urticaria pigmentosa therefore does not appear to be directly related to a change in mast cell numbers or morphology, or to the histamine concentration in the skin. Urticaria pigmentosa usually presents as a generalized maculo-papular rash which urticates on rubbing (Darier's sign). Many patients are troubled only by the unsightliness of the rash while some complain of pruritus, wealing or flushing. These symptoms are attributed to the release of histamine by mast cells which characteristically occur in increased numbers in the dermis. Symptomatic treatment is often unrewarding, but favourable results have been claimed for cimetidine with or without Hi blockers (Hirschowitz & Groarke, 1979; O'Laughlin & Bredfeldt, 1980) and for oral disodium cromoglycate (Soter, Austen & Wasserman, 1979; Czarnetski & Behrendt, 1981). In 1978, Christophers and colleagues reported that photochemotherapy (PUVA) produced symptomatic relief in all of ten adult patients with typical urticaria pigmentosa. Similarly encouraging results were subsequently reported from other centres (Ortonne et al., 1980; Allevato, Donatti & Cordero, 1980; Granerus, Roupe & Swanbeck, 1981; Väätäinen, Hannuksela & Karvonen, 1981). In this study we examined the effects of PUVA in eight adult patients with urticaria pigmentosa. Although PUVA produced a moderately good clinical response in seven out of these eight patients (reduced pruritus, reduced wealing and faded macules) quantitative studies failed to reveal a consistent effect of PUVA on either the mast cell population density or the histamine concentration in both lesional and clinically uninvolved skin. The findings arc discussed in relation to existing information concerning the effect of ultraviolet radiation (U VR) on mast cells and other constituents of the skin.     

Absence of MHC class II antigen on mast cells at sites of inflammation in human skin

Abstract Since the presence of major histocompatibility complex (MHC) antigens has recently been reported on murine and human mast cells under various conditions, we have investigated their expression on mast cells in different types of cutaneous inflammation. Cryostat sections from lesional biopsies of patients with psoriasis, atopic eczema, chronic urticaria, lichen planus, bullous pemphigoid and urticaria pigmentosa were immunohistochemically stained with monoclonal antibodies against MHC class I and class II antigens using a double staining APAAP/toluidine blue methodology. While strongly positive staining with the antibody directed against MHC class I antigens was found on nearly all mast cells in normal skin and in inflammatory dermatoses, reactivity for HLA-DR and HLA-DQ antigens on mast cells could not be detected, except for less than 2% of cells with doubtful staining. Human mast cells therefore probably play no significant rôle as antigen-presenting cells in the conditions studied.     

慢性荨麻疹患者自体血清皮肤试验结果分析

目的 :探讨慢性荨麻疹患者自体血清皮肤试验的意义。方法 :对 4 0例慢性荨麻疹患者以及2 0例正常对照者、1 0例皮肤划痕症、1 0例异位性皮炎患者进行自体血清皮肤试验。结果 :4 0例慢性荨麻疹患者中 1 3例 (32 5 % )自体血清皮肤试验阳性 ,正常对照组和皮肤划痕症及异位性皮炎患者均为阴性 ,慢性荨麻疹患者的自体血清皮肤试验阳性率显著高于正常对照组及皮肤划痕症和异位性皮炎组 (P值均 <0 0 5 )。结论 :自体血清皮肤试验的测定对自身免疫性慢性荨麻疹有一定的诊断意义。     

Effects of local corticosteroids on acute experimental urticaria

Corticosteroids are often used in the treatment of acute or chronic urticaria. However, their effects on mastocyte activation as well as on the histamine-induced dermal oedema remain poorly investigated. The aim of the present study was to investigate the effects of corticosteroids (CS) on the development of acute experimental urticaria induced by prick-tests with histamine and codeine. This experimental model corresponds to the common form of urticaria. CS were administered at the site of the histamine and codeine prick tests in order to test for a direct effect on the development of acute urticaria. Two types of experiments were performed: 1) after a 48-hour period of topical CS application on the forearm, 7 healthy volunteers were skin prick-tested with histamine and codeine simultaneously in duplicate, one series in the pretreated area and the other in a non-treated area. 2) six other volunteers were prick-tested with histamine and codeine on their forearm, in duplicate. Immediately after testing, intradermal methyprednisolone was injected at the site of the prick-tests in the last series. Skin wheal and flare responses were measured after 20 mns and statistically compared with and without CS treatment. Whereas short-term CS topical application did not appear to modify cutaneous reactivity to histamine and codeine, local CS injection was associated with a significant increase in the flare induced by histamine and codeine (respectively + 18 +/- 3% and + 38 +/- 3%; P = 0.05). The wheal tended to be increased after injected CS. In conclusion, these results show that CS are neither able to prevent nor to improve experimental urticaria, i.e. wheal and flare, and even increase the histamine and codeine-induced erythema. That a similar result could apply to patients with chronic urticaria and with systemic CS remains to be studied.     

Association between chronic urticaria and thyroid autoimmunity: a prospective study involving 99 patients

BACKGROUND: The association between chronic urticaria and thyroid autoimmunity has been a subject of debate. However, this link was suggested in studies searching thyroid microsomal antibodies (TMA), which are less sensitive and less specific than anti-thyroperoxidase antibodies, moreover these studies did not measure anti-TSH receptor antibodies, nor did they use a control group. As a consequence, the results of these studies are difficult to interpret. OBJECTIVE: The aim of this study was to determine whether chronic urticaria is statistically associated with thyroid autoimmunity. METHODS: In a prospective case-control study, we compared the frequency of thyroid autoantibodies in 45 patients with chronic urticaria and in 30 healthy adult volunteers; we also compared the frequency of chronic urticaria in 32 patients with thyroid diseases with thyroid autoantibodies and in 22 patients with thyroid diseases without thyroid autoantibodies. Thyroid autoantibodies and thyroid hormones were measured in all the subjects; antinuclear antibodies, rheumatoid factors, complement, IgE were assessed and routine laboratory tests were done in patients with chronic urticaria. Fisher's exact statistics were used to test our hypothesis. RESULTS: The frequency of thyroid autoantibodies was significantly higher in patients with chronic urticaria than in healthy controls (26.7%/3.3%; p < 0.01). All the patients with thyroid autoantibodies had thyroid hormone concentrations within the normal limits. The frequency of chronic urticaria was not significantly different (12.5%/9.1%; p = 0.7) in patients with thyroid diseases with or without thyroid antibodies. The rest of the biological investigations revealed only 1 patient with connective tissue disease. CONCLUSION: This study shows a significant association between chronic urticaria and thyroid autoimmunity, and that tests to detect thyroid autoantibodies are relevant in patients with chronic urticaria, whereas extensive laboratory tests are not.     

Adhesion molecule expression and the inflammatory cell infiltrate in delayed pressure urticaria

We have investigated the kinetics of the leucocyte infiltrate in delayed pressure urticaria (DPU) in relation to the in vivo expression of the cytokine-regulated cell surface adhesion molecules. E-selectin (endothelial adhesion molecule-1. ELAM-1). intercellular adhesion molecule-1 (ICAM-1), and vascular adhesion molecule-1 (VCAM-1). Immunohistochemical analysis was performed on biopsies taken from unchallenged skin, and at 0, 2, 6, 24, 48 and 120 h after weighted rods had been applied to 13 patients with DPU. There was moderate to marked upregulation of E-selectin at 6 and 24 h after application of pressure. At 24 h, more patients showed expression of VCAM-1 on perivascular cells than before pressure. Moderate expression of ICAM-1 was present in some biopsies from both unchallenged and pressure-challenged skin, but there was no clear trend. In DPU, there was a significant increase in the neutrophil count at 2 h after a pressure challenge, with further increases at 6 and 24 h. The median cell counts per high-power field of eosinophils and monocyte/ macrophages increased significantly at 24 h after pressure. Biopsies from four normal controls subjected to an identical pressure challenge showed no detectable changes in adhesion molecule expression or in the cell infiltrate. The findings in four patients with chronic idiopathic urticaria not associated with DPU were qualitatively similar to (but intermediate in severity between) the findings in DPU weals at 6 and 24 h. These results suggest that vascular endothelial activation is an early response to pressure challenge in DPU, and is also present in chronic idiopathic urticaria. The inflammatory infiltrate in DPU differs in intensity, but not in composition, from that in chronic idiopathic urticaria.     

荨麻疹外周血细胞因子表达模式研究进展

荨麻疹的类型多样,发病机制也各不相同.细胞因子在外周血中的水平充分反应疾病的炎症水平及发病机制所牵涉的效应细胞.在急性荨麻疹中,Th2系列因子的高表达证实了急性荨麻疹的过敏性,大量促炎症因子的高表达反映出急性荨麻疹的炎症性.在慢性荨麻疹中,透过外周血细胞因子的表达水平可以发现,不仅Th1/Th2失衡、还有Th17、Tc17和Th9的活化、促炎症因子和趋化因子的广泛激活和非炎症非免疫系统等均参与了慢性荨麻疹的发生发展过程.自体血清皮肤试验不同反应性的慢性荨麻疹的细胞因子表达模式差异也逐渐被证实.除此之外,可诱导性荨麻疹,如寒冷性荨麻疹和延迟压力性荨麻疹等在部分细胞因子表达上也存在差异.