Differential role of lymphocyte function-associated antigens in the activation of nickel-specific peripheral blood T lymphocytes.

The possible role(s) of the adhesion molecules LFA-1 alpha (CD11a), LFA-1 beta(CD18), ICAM-1 (CD54), CD2 (T11, LFA-2), and LFA-3 (CD58) in the in vitro activation of nickel-specific peripheral blood (PB) T lymphocytes was studied. For this purpose, monoclonal antibodies (MoAb) to these markers were used. Both LFA-2 and LFA-3 appeared to be consistently involved, whereas LFA-1 was inconsistently involved. In studies using antigen-presenting cells (APC) isolated from peripheral blood to present nickel, anti-LFA-1 alpha and/or LFA-1 beta MoAb partially inhibited the in vitro activation of nickel-specific T lymphocytes in nine of 42 patients allergic to nickel. In the other 33 patients variable results, ranging from a slight increase or inhibition of proliferation to no inhibition at all, was observed, in particular when different anti-LFA-1 alpha MoAb were added to the cultures. In those patients who showed no inhibition when anti-LFA-1 (alpha and beta) MoAb were added, no inhibition was also observed when a mixture of anti-LFA-1 (alpha and beta) and ICAM-1 MoAb were added to the cultures. Similar results were also obtained using epidermal APC. In control experiments the various anti-LFA-1 (alpha and beta) MoAb effectively inhibited the tetanus toxoid and Con-A induced T-lymphocyte proliferation as well as the spontaneous aggregation of the JY cell line. Anti-CD2 and anti-LFA-3 MoAb strongly inhibited the proliferative responses of nickel-specific peripheral blood T lymphocytes from all 42 patients. These results indicated that the receptor-ligand interaction between CD2 and LFA-3 is essential for in vitro activation of nickel-specific peripheral blood T lymphocytes. This activation, however, does not regularly involve LFA-1 molecules on T lymphocytes. The involvement of LFA-1 in the activation of nickel-specific T lymphocytes correlated positively with high patch test scores to nickel and the disease activity in contact dermatitis patients.

The autologous mixed epidermal cell-T lymphocyte reaction is elevated in psoriasis: a crucial role for epidermal HLA-DR+/CD1a- antigen-presenting cells.

The objective of this study was to determine whether epidermal cells (EC) from psoriasis lesions and uninvolved skin could stimulate autologous T lymphocytes in the in vitro autologous mixed epidermal cell-T lymphocyte reaction (autologous MECLR). The functional role of antigen-presenting cell (APC) subsets was concurrently determined in this reaction. Mononuclear cells and purified T lymphocytes from peripheral blood of psoriasis patients showed a clear proliferative response to autologous unpurified epidermal cells from involved as well as uninvolved skin. The autologous mixed leukocyte reaction (MLR) was not elevated in psoriasis patients. In healthy controls and contact allergy patients, T-lymphocyte proliferation was not observed either in the autologous MECLR or in the autologous MLR. The level of proliferation in the autologous MECLR from psoriasis patients correlated to the number of epidermal cells that were added. To exclude the possibility that the observed proliferation in the autologous MECLR in psoriasis was due to the presence of epidermal T lymphocytes that were being stimulated and expanded in vitro, the stimulator EC were gamma irradiated (30 Gy) in some experiments. Preincubation of EC with cyclosporin A (CsA) significantly inhibited the autologous MECLR. The CsA-induced inhibition could be neutralized by the addition of fresh untreated EC to these cultures. This indicated that one of the modes of action of CsA in resolving psoriasis is, as some investigators have already shown, via inhibition of epidermal accessory cell function. In the autologous MECLR, APC from psoriasis skin could initiate this reaction, whereas APC from peripheral blood could not. This occurred in an MHC class II restricted fashion. Depletion experiments showed that Langerhans cells (HLA-DR+/CD1a+) were not the principal stimulators of autologous T lymphocytes in the MECLR. These results indicated that mainly HLA-DR+/CD1a- epidermal cells from psoriasis patients could stimulate autologous peripheral blood T lymphocytes in an MHC class II-restricted fashion.

Anti-Mycobacterium leprae monoclonal antibodies cross-react with human skin: an alternative explanation for the immune responses in leprosy.

A panel of 17 mouse monoclonal antibodies (MoAb) raised against Mycobacterium leprae (M. leprae) antigens was used to detect antigenic determinants in normal human skin. An indirect immunoperoxidase technique was used. Eight of the MoAb detected epidermal antigens similar to patterns well known for human sera. Five of these MoAb detected determinants in the dermis, too. These observations may indicate a certain degree of similarity between the antigenic determinants occurring in M. leprae and in the human host. We propose that such a similarity on the one hand may facilitate the survival of M. leprae in the human host when the antigens are not recognized as "non-self," a situation which seems to occur in lepromatous leprosy, when the patients' tissues are loaded with bacteria virtually without any immune response. On the other hand, M. leprae antigens which mimic host antigens may induce an auto-immune reaction against the host's own antigens, which could explain the immune reaction in tuberculoid leprosy and during a "reversal reaction" when M. leprae is not observed in the host tissues, but extensive granuloma formation occurs.

Characterization of the mononuclear infiltrate in basal cell carcinoma: a predominantly T cell-mediated immune response with minor participation of Leu-7+ (natural killer) cells and Leu-14+ (B) cells.

We investigated the peritumoral inflammatory infiltrate in 22 basal cell carcinoma (BCC) from 18 patients using a series of monoclonal antibodies. In all the 22 BCC the infiltrate consisted mainly of T cells (55 +/- 15%) and only in three cases an invasion of the tumor nests by these cells was observed. The T helper (TH) subset predominated over the T suppressor/cytotoxic (TS/C) subset (TH/TS/C ratio of 1.9 +/- 0.8). In 8 of 22 BCC mild infiltrate was observed with 48 +/- 13% T cells and a TH/TS/C ratio of 1.5 +/- 0.6. In 14 of 22 BCC moderate to heavy infiltrate with 59 +/- 15% T cells and a TH/TS/C ratio of 2.0 +/- 1.0 was observed. There was a significant difference in the percentage of T cells in BCC with moderate to heavy infiltrate and that in BCC with mild infiltration. The mean percentage of HLA-DR+ cells was 54 +/- 11%; Langerhans cells (LC) 4 +/- 5%; and Leu-M5+ (monocytes and macrophages) 16 +/- 11%. Less than 2% Leu-14+(B) cells were seen in the infiltrate. The mean percentage of Leu-7+ (natural killer) cells was 4 +/- 4%, and only 1 of 22 BCC Leu-7+ cells invaded tumor nests, contacting with tumor cells. From these results we concluded that T cells play a major role in the defence against BCC proliferation. The main role of Langerhans cells and Leu-M5+ cells may be that of antigen presentation. B cells and NK cells probably play a minor role in the local defence against BCC proliferation.

Interleukin-1 and interleukin-6 in psoriasis.

We report on the levels of expression of IL-1 and IL-6 in skin from psoriasis patients. Different approaches were pursued. Initially, the levels of IL-1 beta and IL-6 were measured in suction blister fluid from lesional and uninvolved skin from psoriasis patients, using a sensitive enzyme-linked immunoabsorbent assay (ELISA) and bio-assay. Skin sections were also examined for the presence of IL-1 and IL-6 using IL-1 beta- and IL-6-specific antibodies. Finally, the expression of IL-1 and IL-6 mRNA was determined in cultured keratinocytes (KC) and fibroblasts from psoriasis skin. Suction blister fluid from lesional and uninvolved psoriasis skin and from skin of healthy individuals did not contain detectable levels (greater than 100 pg/ml) of IL-1 beta. Blister fluid from psoriasis lesions contained low but significant levels of IL-6, whereas the serum levels of IL-6 in these patients was undetectable. Using cryostat skin sections and an IL-1 beta-specific monoclonal antibody (MoAb) in an indirect immunoperoxidase technique, a diffuse staining in the entire epidermis was observed in sections of uninvolved skin from psoriasis patients. In cryostat sections of psoriasis lesions, a faint diffuse staining of the epidermis and a pronounced "dot-like" intracellular staining pattern was observed. On the other hand, the same IL-1 beta-specific MoAb showed, in a indirect immunofluorescence technique using unfixed epidermal cells, bright membrane staining in epidermal cell suspensions from psoriasis lesions. Slightly elevated levels of IL-1 beta and IL-1 alpha mRNA were observed in cultured KC from psoriasis lesions as compared to those in normal KC and in the HEp-2 cell line. Very low levels of IL-6 mRNA were expressed in KC from psoriasis lesions and healthy individuals. Fibroblasts from psoriasis lesions expressed extremely low levels of IL-1 alpha and IL-1 beta, but high levels of IL-6 mRNA. The results point to a paradoxical situation in psoriatic skin: blister fluid from psoriasis lesions contains no IL-1 beta, whereas IL-1 beta is overexpressed on the plasma membrane and in the intracellular compartment of epidermal cells. This finding may help in explaining the observed absence of IL-1 in aqueous extracts of psoriatic scales. Because cultured KC from psoriasis lesions express minimal levels of IL-6 mRNA. dermal fibroblasts, probably together with the inflammatory infiltrate, may represent a major source of IL-6 in psoriasis lesions in vivo.

Expression of E-cadherin, alpha- & beta-catenin, and CD44V6 and the subcellular localization of E-cadherin and CD44V6 in normal epidermis and basal cell carcinoma.

Basal cell carcinoma (BCC) of the skin is a locally invasive, rarely metastasizing epithelial tumor. In the current study, the expression of E-cadherin, alpha- and beta-catenin and CD44V6 in normal epidermis and on BCC cells were investigated. A significantly reduced expression of alpha-catenin and CD44V6 and a slightly reduced expression of E-cadherin on BCC cells were observed compared with the overlying epidermis. Immunoelectron microscopy was used to investigate whether the decreased expression of E-cadherin and CD44V6 was due to either an absence or downregulation of specific membrane structures or due to an overall downregulation of these adhesion molecules in all membrane structures in BCC. E-cadherin and CD44V6 were expressed in adherens junctions, desmosomes, and complex interdigitating membrane structures both in normal epidermis and in BCC. A quantitative analysis showed that only a percentage of desmosomes was stained. In addition, the effect of pro-inflammatory cytokines, such as interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), was investigated in biopsy specimens of normal skin and BCC, using a biopsy culture system and immunohistochemistry. The expression of E-cadherin and CD44V6 was not significantly decreased after culturing BCC or normal skin biopsy specimens for 48 hours with or without recombinant human (rHu)IFN-gamma or rHuTNF-alpha. It may be concluded that the decreased expression of both E-cadherin and CD44V6, observed in light microscopy, was not attributable to the absence of specific specialized structures in BCC and most likely also not caused by downregulation by local cytokines, but rather by generic downregulation of both of these adhesion molecules during malignant transformation.

Replication characteristics and core size of intranuclear herpes simplex virus (HSV-1) in genital skin lesions: electronmicroscopy studies of a biopsy from a female patient.

Herpes simplex virus (HSV) type 1 genital infection, leading to ulcerating lesions in a female patient, was studied by electronmicroscopy. Infection had probably been recent, through oro-genital contact with a cold sore on the husband's lip. Cell-culture typing and serological tests indicated that the patient currently had an HSV-1 secondary infection. Aspects studied in a skin biopsy from an ulcerating labium majus were epidermal cell types infected, stages in virus genesis, virus core diameter in intranuclear capsids and extracellular appearance of virus. Different stages in virus genesis, in virus envelope formation and in nuclear and cytoplasmic degeneration were observed in the few remaining, rounded and swollen, epidermal (?) spinosum cells. Their nuclei, some with marginated chromatin, harboured besides dense-cored or empty capsids, electron-dense blobs possibly representing clones of immature virus and falling apart into aggregates of small granules. In other nuclei, large clusters of dense-cored capsids, some distinctly hexagonal in shape, had accumulated in wide gaps in the nuclear membrane whereas remaining nuclear membrane portions were quadruple and often engaged in viral envelope formation. Partially enveloped capsids and naked dense-cored capsids were seen extracellularly indicating their survival outside cells. An occasional virion was present in dermal blood vessel lumina. Measurements of the electron-dense core (nucleoid) of intranuclear capsids in electronmicrographs showed that the HSV-1 core diameter differs very significantly from the core of intranuclear HSV-2 capsids, thus allowing a clear distinction by electronmicroscopy between the two HSV subtypes in plastic-embedded biopsies.

The allergens in cosmetics.

The ingredients responsible for allergy to cosmetics were determined in 119 patients suffering from cosmetic-related contact dermatitis. Most reactions (56.3%) were caused by skin care products, followed by nail cosmetics (13.4%), perfumes (8.4%), and hair cosmetics (5.9%). Preservatives were most frequently implicated (32.0%), followed by fragrances (26.5%) and emulsifiers (14.3%). By far the most important cosmetic allergen was Kathon CG, (a preservative system containing, as active ingredients, a mixture of methylisothiazolinone and methyl chloroisothiazolinone) reacting in 33 patients (27.7%). Other frequent causes of cosmetic-related contact allergic reactions were toluenesulfonamide/formaldehyde resin in nail hardener and/or nail lacquer (15 patients [12.6%]), and oleamidopropyl dimethylamine, an emulsifier in baby body lotion (13 patients [10.9%]).

Expression of cytokeratin 8 and low molecular weight cytokeratins in human basal cell carcinoma.

The expression of low molecular weight cytokeratins (Cks) 7,8,18, 18 and high molecular weight cytokeratin 10 in 23 basal cell carcinomas (BCCs) was investigated using a panel of 14 different commercially available monoclonal antibodies (MoAbs) with specific anti-cytokeratin activity. Four of these MoAbs were directed against Ck 8. The results showed that Ck 8 was detected in all 23 BCCs using MoAb 4.1.18. Two of the MoAbs showed inconsistent staining for Ck 8 and one of them did not show any staining at all. Cytokeratins 7 and 19 were detected incosistently. Cytokeratins 18 and 10 were not detected in any of the 23 BCCs that were examined. The incosistent observations on the expression of Ck 8 in BCCs in this study could have been due to different epitopes of the different cytokeratins that were detected by the different MoAbs. The results of this study lead to recommendation that whenever possible a panel of different MoAbs directed against the same Ck(s) should be used in order to minimize the risk of obtaining incorrect experimental results.

Absence of cytokeratin 8 and inconsistent expression of cytokeratins 7 and 19 in human basal cell carcinoma.

The expression of the low molecular weight cytokeratins (K) 7, 8, 18, 19 and the high molecular weight cytokeratin 10 in 21 basal cell carcinoma (BCC) was studied using seven different monoclonal antibodies (MoAbs) with specific anti-cytokeratin activity. MoAbs RCK 105 (anti-K7), RPN 1164 (anti-low molecular weight cytokeratins of basic group), Ks 19.1 (anti-K19) and Cam 5.2 (anti-K8, K18, K19) reacted positively but inconsistently in the BCC that were examined. MoAbs 1166 (anti-K8) and RGE53 (anti-K18) did not react at all. MoAb RKSE 60 (anti-K10) did not react with the tumor cells. From the results of this study, it can be concluded that cytokeratins 7 and 19 are expressed in BCC (43% and 71%, respectively), whereas cytokeratin 8 is not expressed.

Expression of cytokeratin 8 in basal cell carcinoma: a comparative immunohistochemical and immunoelectron microscopy study.

The comparative study reported here was undertaken in order to resolve the discrepancies in the detection of cytokeratin (Ck) 8 reported in previous studies. The expression of Ck 8 was compared in 6 basal cell carcinomas (BCCs) using immunohistochemical and immunoelectron microscopic techniques and a panel of 4 different commercially available monoclonal antibodies (MoAbs). The results of this comparative study demonstrated not only that the consistent expression of Ck 8 using one of the MoAbs in immunohistochemistry was confirmed by immunoelectron microscopy, but that the inconsistent expression of Ck 8 observed using two other MoAbs was also confirmed. One of the MoAbs did not show any staining at all. The inability of this MoAb to detect the expression of Ck 8 using either of the techniques also indicated that this MoAb may be directed against an epitope of Ck8 that is not detectable in BCC in situ.

Immunoelectron microscopic studies on cytokeratins in human basal cell carcinoma.

Ultrastructural investigations into the location and the expression of the cytokeratins 7, 8, 10 and 19 were undertaken on ultrathin cryosections of 8 basal cell carcinomas (BCC) using a high resolution immunogold labeling method and five different monoclonal antibodies against specific cytokeratins. The results showed that cytokeratin 10 was expressed only in the differentiated keratinocytes. In contrast to the previously reported biochemical and immunohistochemical studies, in this study cytokeratin 19 was expressed not only in the tumor cells of BCC but also in the normal epidermal keratinocytes. The expression of cytokeratin 7 in BCC could not be confirmed but the lack of expression of cytokeratin 8 was confirmed, excluding its potential role as a specific histopathological marker for BCC.

The recruitment of inflammatory cells using the skin-window technique.

The chemotaxis of inflammatory cells induced by the skin-window technique using IgD as cytotaxigen or cytotaxinogen was studied in 16 patients with allergic contact dermatitis. Six patients with leg ulcer served as controls. By means of this method the recruitment of inflammatory cells with receptors for IgD could be shown.

Adhesion molecules and IL-1 costimulate T lymphocytes in the autologous MECLR in psoriasis.

Membrane molecules such as CD36 (OKM5), intercellular adhesion molecule-1 (ICAM-1, CD54), gamma interferon-induced protein 10 (gamma-IP10) and IL-1 are induced and/or upregulated in psoriatic epidermis. These molecules have important accessory, trafficking or signalling functions in the immune system and also play a role in the pathophysiology of psoriasis. The relevance of adhesion molecules, CD36 and epidermal IL-1 in psoriasis was studied in vitro in the autologous mixed epidermal cell - T lymphocyte reaction (MECLR). Their level of expression was quantitated in epidermal cell suspensions (ECS) from patients with psoriasis and their function was assessed by blocking with specific mAbs and antisera or by depleting CD36+ cells from the ECS prior to the MECLR. ECS from psoriatic lesions contained increased numbers of CD36+ (23 +/- 12%), ICAM-1(+) (31 +/- 14%) and IL-1(+) (57 +/- 21%) cells. The autologous MECLR was inhibited in samples from all patients by mAb to CD2 (LFA-2), CD11a (LFA-1alpha), CD18 (LFA-1beta), ICAM-1, CD58 (LFA-3) and an antiserum to IL-1beta. Thus, adhesion molecules facilitate inflammation in psoriasis not only via adhesion and recruitment of T lymphocyte in psoriatic lesions, but also via activation of T cells. Furthermore CD36 molecules on psoriatic epidermal cells do not costimulate autologous T lymphocytes in psoriasis. The observed costimulatory function of IL-1beta in the MECLR emphasizes its relevance in psoriasis.

Double-blind comparison of 200-mg ketoconazole oral tablets and 1200-mg miconazole vaginal capsule in the treatment of vaginal candidosis.

In a double-blind, randomized study the efficacy of oral ketoconazole (400 mg for 5 days) was compared with that of topical miconazole (1200 mg a single dose) in the treatment of 42 patients with vaginal candidosis. One week after the start of therapy the mycological cure rate was 87% in the ketoconazole group and 95% in the miconazole group, indicating that both regimens are equally effective. No major side-effects were observed.

A comparison of acrivastine versus terfenadine and placebo in the treatment of chronic idiopathic urticaria.

Patients (n = 56) with a diagnosis of chronic idiopathic urticaria were assessed in a fully randomized, double-blind, crossover study to investigate the efficacy of acrivastine at two doses (8 and 4 mg) versus 60 mg terfenadine and placebo administered three times daily. All three active preparations were found to be effective, and significantly better than placebo, in controlling the signs and symptoms of urticaria. No significant differences were found between the active preparations, although in some cases efficacy trends favoured 8 mg acrivastine and terfenadine over 4 mg acrivastine. No significant differences were noted between the active treatments and placebo with regard to reports of drowsiness.

[Epoxy resin as a cause of contact allergic eczema]. / Epoxyhars als oorzaak van contactallergisch eczeem.

Epoxy resin-containing products of the plastic industry are increasingly employed. We retrospectively studied the incidence and course of epoxy resin contact dermatitis in 2265 patients in whom contact dermatitis was confirmed by patch testing. A positive patch test for epoxy resin was present in 31 patients (1.4%). Exposure to epoxy resin as the cause of contact dermatitis was 'certain' in 10, 'very likely' in 9, 'possible' in 4 and 'unclear' in 8 patients. Exposure to epoxy resin was related to occupational (55%) and leisure time activities (13%). In 48% patch tests were positive for multiple test allergens. Hands (56%), arms (38%) and face (38%) were mainly affected. Re-examination of 26 of 31 patients showed that a favourable course depended on identification of the cause of the exposure to epoxy resin, avoidance of further contact and absence of sensitization to multiple allergens.

[Parapemphigus and variants; clinical and immunological aspects]. / Parapemphigus en varianten; klinische en immunologische aspecten.

The clinical, histopathological and immunological findings and therapeutic data of 29 patients with different variants of pemphigoid seen by us in the period 1981-1988 are summarized. All 29 patients: disseminated or bullous pemphigoid (20 cases), localized pemphigoid (3 cases), cicatricial pemphigoid (2 cases), juvenile pemphigoid (2 cases), papular pemphigoid (1 case) and pemphigus vegetans (1 case) showed as common denominators subepidermal blister formation and linear deposition of Ig (IgG) and complement in the basement membrane zone. In nearly all the cases with disseminated pemphigoid treatment with immunosuppressive drugs orally was indicated. With particular reference to the age of patients in this subgroup (mean age greater than 70 years) aspects inherent to the treatment are briefly discussed. In contrast to the (relatively) benign course of the disease in localized pemphigoid and in juvenile pemphigoid, cicatricial pemphigoid with mucosal localization appears to be a variant within the spectrum of pemphigoid with severe complications due to tissue retraction and loss of local function.

[Contact allergy for Alstrumeria (inca lily)]. / Contactallergie voor Alstroemeria (incalelie).

We report 6 patients with occupational contact allergy to Alstroemeria cultivars. Four of them presented with the clinical picture of 'tulip fingers'. They all reacted to parts of fresh plants and to tuliposide A. The literature on Alstroemeria allergy is reviewed.

[Is juvenile seborrheic dermatitis a candidiasis? Studies of a possible link with microbial infections]. / Is juveniele seborrhoïsche dermatitis een candidide? Onderzoek naar mogelijk verband met microbiële infecties.

Infantile seborrheic dermatitis (ISD), a disease occurring in the first months of life, is an erythematosquamous skin disease of unknown origin. This article presents results of microbial studies in 20 patients with ISD. Isolation of candida in a high percentage may indicate a primary role of this micro-organism in the etiology of this disease. It is striking that this disease often starts after disturbing the microbial colonization of the intestinal tract. Often ISD develops during the transition of breastfeeding to humanized cow milk.