Preliminary evaluation of in vivo reflectance confocal microscopy features of Discoid lupus erythematosus.

BACKGROUND: Discoid lupus erythematosus (DLE) can simulate other inflammatory diseases both clinically and histologically. In vivo reflectance confocal microscopy (RCM) is a noninvasive, reproducible imaging technique already reported to be useful in the evaluation of several inflammatory skin conditions such as contact dermatitis, psoriasis and Darier disease. OBJECTIVES: The aims of our study were to define RCM features of DLE and to evaluate its feasibility in biopsy site selection. METHODS: Discoid lesions were selected for RCM evaluation from 10 patients with an established diagnosis of DLE. Subsequently, a 4-mm punch biopsy of the same areas evaluated with RCM was rendered for histopathological examination. RESULTS: A series of RCM features of DLE was identified and shown to correlate well with histopathological evaluation. Interface changes, as well as epidermal, dermal and adnexal inflammatory cell infiltration, were identified with RCM in a high percentage of the lesions. A limitation of RCM examination besides imaging depth was the inability to distinguish lymphocytes from other white blood cells. CONCLUSIONS: The utility of RCM as a diagnostic tool for DLE awaits further evaluation, although it appears to be promising for biopsy site selection.

Nerve growth factor and asthma.

An increasing body of evidence shows that nerve growth factor (NGF) exerts biological activity not only on the central and peripheral nervous system, but also on the immune system thereby influencing allergic diseases and asthma. (1) NGF circulating levels are increased in patients with allergic diseases and asthma, and are related to the severity of the inflammatory process and disease. In vernal keratoconjunctivitis, NGF plasma levels correlate with the number of mast cells infiltrating the conjunctiva, and NGF mRNA is increased in nasal mucosal scrapings of patients with allergic rhinitis who have high levels of NGF in serum and nasal fluids; NGF is further increased in nasal fluids after specific allergen challenge. (2) NGF is produced and released by several modulatory and effector cells of allergic inflammation and asthma, for example T-helper 2 lymphocytes, mast cells and eosinophils. (3) NGF receptors are expressed on the conjunctival epithelium of patients with allergic conjunctivitis and the number of NGF-receptor positive cells is increased in the conjunctiva of these patients. Indeed, local administration of NGF induces fibroblast activation and healing processes of human corneal ulcers, which suggests that NGF plays a role in tissue remodelling processes occurring in asthma. (4) NGF increases airway hyperreactivity to histamine in an animal model of asthma, while anti-NGF treatment reduces airway hyperreactivity induced by ovalbumin topical challenge in the sensitized mouse.

Use of altered peptide ligands to modulate immune responses as a possible immunotherapy for allergies.

Allergies are dramatically increasing in prevalence, and the management of these diseases is a heavy burden on the health-care systems of developed countries. In recent years, many efforts have been made to improve the therapy of allergies and to develop new approaches for immunotherapy. Here we briefly review the use of peptides to modulate T-cell responses to allergens. We focus mainly on the possibility of using altered peptide ligands (APLs), i.e., peptides tailored on immunodominant T epitopes and bearing a single amino-acid substitution, as a tool to modulate immune responses to allergens. These peptides may be recognized by the specific T cells triggered by the agonist peptides, but they are unable to elicit T-cell responses; thus, they could be ideal candidates to modulate immune responses to allergens. The availability of these peptides could allow new approaches for immunotherapies.

Immunological findings in thymoma and thymoma-related syndromes.

Human thymoma is a neoplasm of thymic epithelial cells associated with several clinical syndromes ranging from autoimmune disease to immunodeficiency. The aim of our research was to investigate T cell-mediated immune response in patients with thymoma. Initially eight patients were enrolled in this study. Four patients underwent surgical removal of the thymus, while four were submitted to diagnostic procedures only. Inversion of the CD4:CD8 ratio was found in three patients. Only one subject displayed a normal CD19 count in peripheral blood. The mean value (+/-SD) of the CD19 percentage in the patient group was 2 +/- 2.2. Notably, the patients with thymoma had fewer mature B lymphocytes than the thymectomized patients. The T-cell receptor (TCR) repertoire was investigated in three individuals affected by thymoma: one underwent thymectomy, while the two others, one of which presented with lymphocytosis, were submitted to diagnostic biopsies only. The preliminary results showed a marked alteration in the CD8 repertoire of the thymectomized patient but not in that of the lymphocytotic patient. However, alterations in the TCR repertoire were also found in one patient with thymoma. Altogether, these preliminary findings reveal that loss of CD19+ lymphocytes in peripheral blood is a frequent phenomena in thymoma patients. In this article we discuss this aspect in the context of alterations of the TCR repertoire.

Use of antagonist peptides to inhibit in vitro T cell responses to Par j1, the major allergen of Parietaria judaica pollen.

Antigenic peptides with substituted side chains inhibit immune responses to a number of recall Ags from infectious agents in vitro. Here we show that the same strategy can be applied to peptides derived from a pollen protein, the major allergen of Parietaria judaica(Par j1), a plant responsible for most allergenic sensitization in the southern Mediterranean area. Three T cell lines responding to Par j1 protein were used to identify a stimulatory peptide. Two different monosubstituted altered peptide ligands (APL) were identified that bound to the HLA-DR of the responders, did not stimulate the T cell lines on their own, and decreased the response to subsaturating amounts of the unmodified stimulatory peptide. Most important, these APL were able to inhibit the response of these cell lines to intact Par j1 protein. A third monosubstituted peptide bound to the HLA-DR but did not show inhibitory activity. The two APL had a lower affinity than the unsubstituted peptide for the HLA-DR. The last two observations make MHC blockade an unlikely explanation for the observed effect. These results indicate the action of a specific peptide-mediated antagonism that may be useful in controlling the T cell component of an allergic response.

HLA-DRB1* and allergy to Parietaria: linkage and association analyses.

In this study, we used the affected sibling-pairs approach to investigate the linkage of HLA (human leukocyte antigen)-DRB* with phenotypes related to allergy to Parietaria, the most common pollinosis in Mediterranean countries. The study population consisted of 51 nuclear families (235 subjects). Linkage was detected with Parietaria skin test positivity (p < (0.01), presence of IgG and IgE antibodies specific for the major allergen Par o 1 (p < 0.020 and p < 0.025, respectively), and absence of Par o 1-specific IgE (p < 0.020). High levels of Par o 1-specific IgG were associated with DRB1*1101 and/or DRB1*1104 (p < 0.0001 and p < 0.0119, respectively) in parents and probands. High levels of Par o 1-specific IgE were associated with DRB*1104 in parents (p < 0.017) and with DRB1*1101 in probands (p < 0.0146). When siblings were categorized according to high/low total IgE levels (> or =125 IU/ml and <125 IU/ml, respectively), high IgE antibody response was associated with DRB1*1104 in siblings with low total IgE (p < 0.034) and with DRB1*1101 in siblings with high total IgE (p < 0.05). These results demonstrate that HLA-DRB1*, or genes in linkage disequilibrium, contributes to susceptibility to Parietaria allergy and that total IgE levels can discriminate population subsets where different alleles (at the HLA region or at loci in linkage disequilibrium) contribute to control allergen-specific IgE synthesis.

Association of responsiveness to the major pollen allergen of Parietaria officinalis with HLA-DRB1* alleles: a multicenter study.

Parietaria, a plant belonging to the family of Urticaceae, is a major source of allergenic pollen in Europe. In the context of a multinational study, we investigated whether in allergic subjects antibody response towards Par o 1, the major allergen from P. officinalis, was associated with defined HLA-DRB1* alleles. The study population consisted of 234 allergic patients: 65 from Bulgaria, 30 from Israel, 99 from Italy, and 40 from Spain. In the Italian study group, the prevalence of ST positivity to Parietaria was 77%. In Parietaria ST-positive subjects, the prevalences of IgG and IgE serum Ab towards Par o 1 were 91% and 75%, respectively. HLA-DRB1*1101 and/or 1104 were significantly positively associated with the presence of IgG Ab and with high levels of IgE Ab towards this allergen (p = 0.0007 and p = 0.012, respectively). In the Spanish study group, the positive association of DR1100 with responsiveness to Par o 1 was confirmed (p = 0.02, RR = 4, and p = 0.002, RR = 7, for IgG and IgE Ab, respectively). None of the Bulgarian patients had IgE Ab to Par o 1, whereas IgG Ab response was observed in 7 out of 65 subjects and was positively associated with DRB1*1101 and/or 1104 (p = 0.025). In the Israeli study group, responsiveness to Par o 1 was not associated with specific HLA-DRB1* alleles. In conclusion, this study shows that in allergic patients from three European populations antibody response to the major allergen from the pollen of Parietaria is associated with HLA-DRB1*1101 and/or 1104. Our data suggest that this association is stronger in subjects monosensitized to Parietaria.

[Weary hereditary sclerosing poikiloderma]. / Poïkilodermie sclérosante héréditaire de Weary.

INTRODUCTION: Hereditary sclerosing poikiloderma is a genodermatosis with dominant autosomal transmission and variable penetration. The first case was described by Weary in 1969 in 7 members of two black families. CASE REPORT: A 10-year-old girl had localized regional poikiloderma of the fingers and club toes. These lesions were associated secondarily with linear symmetric bands of sclerotic tissue in the axiallary regions. On the X-ray examinations of the distal phalanges of the fingers and the toes showed a proximal growth foyer and absent ungueal phalanges, excepting in the fourth finger of the left hand. Capillaroscopy of the supra-ungueal fold of the fingers showed abnormal capillary circulation. Histology and ultrastructural examinations did not reveal any pathognomonic alterations. DISCUSSION: This case is the first reported in a white patient. The radiological aspect and the results of the capillaroscopy of the fingers and the toes have not been reported previously in this rare genodermatosis. Inheritance of this genodermatosis is poorly defined.

Soluble E-selectin and soluble tumour necrosis factor receptor (60 kD) serum levels in patients with psoriasis.

BACKGROUND: Increased tumour necrosis factor alpha has been found in psoriatic skin. This cytokine activates endothelial cells and induces the membrane E-selectin molecule (E-selectin or endothelial leucocyte adhesion molecule 1); the same cytokine is able to induce its own receptors. Since the soluble forms of E-selectin and tumour necrosis factor receptor (TNF-R, 60 kD) may be reliably measured in body fluids, these determinations have been performed in the sera of psoriatic patients. OBJECTIVE: To evaluate endothelial activation in psoriatic patients, sE-selectin has been determined in patient sera and compared with those of a control group. sTNF-R (60 kD) was also measured in the same samples. METHODS: Two commercially available enzyme immunoassay methods have been used to determine sE-selectin and sTNF-R (60 kD) in the sera of 19 patients with plaque-type psoriasis; 22 healthy subjects were used as controls. CONCLUSIONS: Significantly increased amounts of sE-selectin serum levels were found in psoriatic patients as compared to healthy controls. Moreover, a direct correlation between sE-selectin and PASI scores was observed. On the contrary, sTNF-R (60 kD) serum levels presented no increases. These data suggest that sE-selectin serum levels are a reliable marker of disease activity in psoriatic patients.

IL-10 levels are decreased in psoriatic lesional skin as compared to the psoriatic lesion-free and normal skin suction blister fluids.

IL-10 is a cytokine produced by B and T-cells, monocytes and keratinocytes with pleiotropic effects, some of which are directed towards suppressing monocyte activities (anti-inflammatory cytokine). No information at the protein level is available concerning IL-10 in suction blister fluids from psoriatic skin, even if contrasting data have been reported on IL-10 mRNA of psoriatic biopsies and on the cytokine patterns of the T-cell clones, isolated from psoriatic skin. The IL-10 blister fluid concentrations in psoriatic lesions were compared to those found in the non-lesional skin of 14 patients effected with plaque-type psoriasis, and to those found in the skin of healthy controls (9 subjects sharing sex ratio and age with psoriatic patients). No difference in the IL-10 levels was found between non-lesional and control skin. In contrast, lower IL-10 levels were observed in blister fluids obtained from lesional psoriatic skin (p < 0.0005). The possible meanings of these results have been evaluated in the context of the mechanisms activating or maintaining the chronic inflammatory components of psoriasis.

Correlated increases of tumour necrosis factor-alpha, interleukin-6 and granulocyte monocyte-colony stimulating factor levels in suction blister fluids and sera of psoriatic patients--relationships with disease severity.

Tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and granulocyte monocyte-colony stimulating factor (GM-CSF) were measured in serum and involved and uninvolved skin blister fluids of 20 psoriatic patients and 10 healthy subjects, by enzyme immunoassay. TNF-alpha and IL-6 were always detectable in involved skin blister fluids, while GM-CSF was detected only in 45% of these samples. TNF-alpha, IL-6 and GM-CSF were detected in 95, 100 and 10% of uninvolved skin blister fluid samples, respectively. TNF-alpha and IL-6 were found in 50 and 30% of control blister fluids, while GM-CSF was never detected. In serum, TNF-alpha was detected in 75% of patients and in 70% of controls; IL-6 in 45% of patients and in no controls; and GM-CSF in 35% of patients and in 20% of the controls. The median TNF-alpha and IL-6 levels in involved skin were statistically higher than those of both uninvolved and control skin blister fluids. TNF-alpha and IL-6 levels in blister fluids obtained from both involved and uninvolved skin were higher than those of the patients' sera. GM-CSF, when present in involved skin blister fluids, showed correlated levels with the other cytokines (TNF-alpha: R = 0.85, P = 0.004; IL-6: R = 0.72, P = 0.03). TNF-alpha was highly correlated with IL-6 (R = 0.78, P < 0.00001) in involved skin blister fluids. TNF-alpha and IL-6 levels of involved skin blister fluids showed significant correlations with the psoriasis area and severity index scores in the patients, suggesting a direct relationship between these cytokines and the clinical manifestations of the disease. Moreover, the TNF-alpha levels were particularly related to the erythema scores in the patients, further supporting evidence of their role in the pathogenesis of the disease.

Effect of telenzepine, an M1-selective muscarinic receptor antagonist, in patients with nocturnal asthma.

Muscarinic M1-receptor antagonists can prevent the induction of a long-lasting excitatory postsynaptic potential in autonomic ganglia. As the prolonged occupation of M1-receptors is a possible protective mechanism against vagal overstimulation, M1-antagonists might prove to be effective in preventing nocturnal wheeze. The aim of the present study was to investigate whether telenzepine, an M1-selective muscarinic receptor antagonist, administered orally at different doses (1.5 mg, 3 mg, 5 mg) for one week, reduced airway obstruction in patients with nocturnal asthma in comparison with placebo. Peak expiratory flow rate (PEFR) difference between each study medication (arithmetic means from values on days 4, 5 and 6) and baseline at 6 a.m. and at midnight on day 5 did not differ significantly. Treatment with placebo nocte (at night, 9 p.m.), or telenzepine 1.5 mg nocte, telenzepine 3 mg nocte and telenzepine 5 mg nocte did not affect significantly the 24-h time course of PEFR. The results of this study indicate that the use of telenzepine via the oral route at a dose of up to 5 mg is not effective in preventing nocturnal asthma.

Interleukin-1-beta, interleukin-6, and interferon-gamma in suction blister fluids of involved and uninolved skin and in sera of psoriatic patients.

Interleukin-1-beta (IL-1-beta), Interleukin-6 (IL-6) and Interferon-gamma (IFN-gamma) were measured by enzyme immunoassay (EIA) methods in blister fluids (BFs) obtained from both involved (ISBF) and non-involved skin (USBF) and in sera from 14 psoriatic patients. The same determinations were carried out in 14 sera and in 5 suction blister fluids from 14 normal subjects. IL-6 was always detectable in all skin fluids and in 3 psoriasis sera. IL-1-beta was measured only in 5 ISBFs and in 5 sera from the same patients. IFN-gamma was present in 11 ISBFs, in 5 USBFs and in 5 sera. The analysis of the levels found in the samples shows: 1) a local production of these cytokines, 2) the presence of detectable amounts of IL-6 and IFN-gamma in USBFs, and 3) a significant correlation between the IL-6 levels in the ISBFs and erythema score.

Soluble intercellular adhesion molecule-1 and procollagen III peptide are reliable markers of disease severity in psoriasis.

Levels of soluble intercellular adhesion molecule-1 (sICAM-1) and procollagen III peptide (PIIIP) were measured respectively by enzyme immunoassay (EIA) and radioimmunoassay (RIA) methods in sera from 14 patients affected with psoriasis. The same determinations were also performed on suction blister fluids (BFs) obtained from lesional and non-lesional skin. Fourteen normal subjects were used as controls. Significant correlations were found between the serum levels and psoriasis area and severity index (PASI), (R = 0.62 for sICAM-1 and R = 0.73 for PIIIp, respectively). Of the PASI components, infiltration and erythema represented the variables most closely related to PIIIP (R = 0.85; R = 0.72 respectively). Differently from PIIIP, whose levels were significantly lower in the sera than in skin BFs (serum: median value 1.05, range 0.7-2.3 vs. lesional skin fluid: 11.8, 4.8-30 U/ml), sICAM-1 molecules were found predominantly in the sera (serum: median 316, range 117-579 vs lesional skin fluid: median 70, range 31-252 ng/ml). These data cannot exclude that sICAM-1 molecules detected in suction BFs may derive from serum contamination.

[Microparticulate aerosols and respiratory allergy]. / Microparticolato aerosolico e allergopatie respiratorie.

The assessment of allergenic activity in submicronic particles could explain some unknown aspects of pollinosis pathogenesis. Twenty-five 0.5 discs have been obtained using a high volume sampler (Hi Vol Andersen) equipped with 0.3 micron Whatman paper filters. These discs have been challenged with a concentrated pool of sera of Parietaria allergic patients by RIA in order to evaluate the presence of allergenic activity on filters. Discs of non sampled filters and discs of sampled filters challenged with serum pool of patients sensitized to house dust mites were used as controls. The percentage of bound radioactivity was detected by gamma-counter. The radioactivity bound to sampled discs with Parietaria sera was 2.3 +/- 0.55 (Standard Deviation); radioactivity detected on control discs was comparable to background values detected by counter. These preliminary date might suggest that submicronic particles of Parietaria can retain some allergenic activity.