Progesterone sensitive Interferon-gamma producing cells detected by ELISpot assay in autoimmune progesterone dermatitis.

Autoimmune progesterone dermatitis (APD) is a rare cutaneous disorder, characterized by recurrent polymorphous cutaneous and mucosal manifestations. It is considered to be caused by a hypersensitive reaction to endogenous progesterone. However, in vitro T-cell activity to this hormone has been described in few patients. Here we report the case of a 30-year-old woman with recurrent pruritic erythematous, and erythema multiforme-like eruptions localized to the genital area. Positive cutaneous reaction to intradermal progesterone injection suggested the diagnosis of APD. The analysis of cellular immune response to progesterone, investigated by the ELISpot assay, showed a significantly higher level of Interferon-gamma (IFN-gamma) producing cells in this patient compared with a control group comprising five asymptomatic women in the luteal phase of the menstrual cycle. Our results suggest that the ELISpot technique, together with clinical evaluations and assessment of allergies, could be useful in the diagnosis of APD.

Increased tryptase and myeloperoxidase levels in blister fluids of patients with bullous pemphigoid: correlations with cytokines, adhesion molecules and anti-basement membrane zone antibodies.

Tryptase and myeloperoxidase respectively represent 2 specific markers of activated mast cells or neutrophils. Therefore, establishing the levels of these enzymes may be useful to quantify the cell involvement in the tissues or fluids of different origins and in different pathologies. The aim of this study was to analyse the levels of these 2 markers in both the sera and blister fluids of patients affected with bullous pemphigoid. These levels were then correlated to the concentrations of 19 cytokines and 2 soluble adhesion molecules determined in the same samples and also with the log (anti-basement membrane zone antibody) titres, evaluated in the patients' sera. For these purposes, 15 patients with bullous pemphigoid (10 males and 5 females; median age: 84 years, range 66-87; median disease duration: 0 years, range 0-3: median number of skin lesions: 17, range 14-30; median anti-basement membrane zone antibody titre: 1:320, range 0.0-1:2560) and 15 normal subjects (11 males and 4 females, median age: 81 years, range 59-86) were analysed by means of commercially available kits. Results showed that blister fluid myeloperoxidase and tryptase levels were increased as compared with the respective sera (P<0.01) and several correlations were observed with cytokines and adhesion molecules. In fact, significant correlations of blister fluid tryptase levels were observed with IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, VEGF, RANTES and sICAM-1, while myeloperoxidase was correlated with IL-1beta, IL-13 and IL-15. The blister fluid tryptase levels were also significantly correlated with the anti-basement membrane zone antibody titres (R=0.53, P=0.05). In conclusion, these findings are in accord with an involvement of both mast cells and neutrophils in bullous pemphigoid and their recruitment may be mediated by different biological modulators. Our findings seem to indicate that the cytokine (IL-3, IFN-gamma and OSM) or adhesion molecule (sICAM-1) concentrations in blister fluid are logarithmically related to the anti-basement membrane zone antibody titers.

Diagnostic relevance of polymerase chain reaction technology for T. pallidum in subjects with syphilis in different phases of infection.

Although serology is a valid tool for the clinician to manage syphilis infection, there are still some cases in which evidence of the presence of T. pallidum or its specific components, such as specific DNA segments, may be useful to establish or confirm the diagnosis. In the absence of T. pallidum grown in culture, a nested PCR to amplify a specific segment of the microorganism genome was performed in ulcerative secretions or sera, after DNA extraction, using a commercially available kit. A kit validation was based on the observation of no positivities in patients without ongoing or anamnestic infection (40 patients). On the contrary, patients infected with T. pallidum presented positivities both in ulcerative secretions and in sera with frequencies that depended on the disease phase and type of sample. In fact, even after treatment, ulcerative secretions that were negative in dark-field examination were found to be positive in PCR. In addition, the sera of patients with positive specific IGM (serologically diagnosed syphilis, asymptomatic state) were also positive in PCR. This test could, therefore, be useful to analyze difficult situations, especially when a seropositivity for a previous infection may complicate the serology of a reinfection or when therapies interfere with dark-field microscopic observation.

Urticaria from beer: an immediate hypersensitivity reaction due to a 10-kDa protein derived from barley.

BACKGROUND: Urticaria from beer has been reported in atopic patients. In these subjects, the skin-prick test positivity to and presence of specific serum immunoglobulin (Ig)E for barley malt, the basic ingredient used in brewing, suggested a type I hypersensitivity to barley component(s). OBJECTIVE: To identify the beer allergen(s) and to investigate the presence of related proteins in barley. METHODS: Three patients with urticaria from beer and other atopic people, some of them suffering from baker's asthma, were examined for both prick test sensitivity to and the occurrence of serum-specific IgE for partially purified proteins from beer. Allergen identification in beer, malt and barley was performed by immunoblotting. RESULTS: Skin-prick tests and detection of specific IgE by both solid-phase (RAST) and liquid-phase (AlaSTAT) assays demonstrated that the 5-20-kDa beer protein fraction contained the allergen. Immunoblot analysis with sera of patients with urticaria from beer showed that IgE bound only the 10-kDa protein band in beer and malt, whereas a main 16-kDa protein was revealed in barley in addition to a very faint 10-kDa band. With the serum of a patient suffering from baker's asthma no IgE binding bands were observed in beer, whereas specific IgE binding to several proteins, including a major 16-kDa component, were detected for both malt and barley. CONCLUSIONS: Urticaria from beer is an IgE-mediated hypersensitivity reaction induced by a protein component of approximately 10 kDa deriving from barley. This allergen does not seem to be related to the major barley 16-kDa allergen responsible for baker's asthma. Because of the severity of the allergic manifestations to beer we recommend testing atopic patients positive to malt/barley and/or who exhibit urticarial reactions after drinking beer for their sensitivity to this beverage.

The serum levels of sE-selectin are increased in patients with bullous pemphigoid or pemphigus vulgaris. Correlation with the number of skin lesions and recovery after corsticosteroid therapy.

Soluble E (sE)-selectin represents the soluble isoform of cellular E-selectin, an adhesion molecule synthesized only by endothelial cells. As a consequence, it may be considered a marker of endothelial activity. The aim of this study was therefore to evaluate the serum levels of sE-selectin in nine patients affected with pemphigus vulgaris (PV) and in 15 patients with bullous pemphigold (BP). Higher amounts of sE-selectin, median 40.3 ng/mL, range 30-109.6 were found in the patients when compared with 20 healthy individuals, median 28.5 ng/mL, range 6.4-48; P < 0.01, matched for sex and age. These levels were also significantly correlated with the number of detectable lesions (r = 0.63, P < 0.001) when the patient data were considered at the time of the first observation. Thirteen subjects were followed over time for a maximum of 3 months (from three to seven observations). During therapy, the number of lesions and the serum sE-selectin values decreased concomitantly. Differently from sE-selectin, the serum soluble intercellular adhesion molecule-1 (sICAM-1) values were not significantly different in the patients from the controls and showed no correlation with the serum sE-selectin concentrations or with the number of lesions. The data presented point to the possible use of sE-selectin determinations as a non-specific follow-up marker, suitable to gauge disease intensity over time and emphasize that endothelial activation is present in BP as well as in PV.

Effect of data normalization for age on the correlations between corneometric values and serum molecule levels in plaque-type psoriatic patients.

Corneometry has been considered useful both to evaluate disease severity and to monitor psoriatic patients during treatment. However, a limitation of this technique is that the patient's age influences the corneometric determinations, thus reducing their clinical usefulness. The aim of this study was, therefore, to establish whether age normalization of the corneometric results may provide more reliable data for clinical use. Corneometric levels were determined in 10 plaque-type psoriatic patients, under standard conditions. Eight serum variables, including transforming growth factor-beta 1 and seven soluble membrane molecules, were assayed with commercially available immune-enzyme methods in the same patients, whose age and PASI scores were also recorded. The age normalization procedure improved all the correlation coefficients calculated on the lesional or non-lesional corneometric values versus the PASI scores as well as versus the other serum variables. This approach may render corneometric determinations more useful to evaluate disease status or treatment effect in patient groups with plaque-type psoriasis.

Use of discriminant analysis to assess disease activity in pulmonary tuberculosis with a panel of specific and nonspecific serum markers.

Several cell activation markers, acute phase reactants, enzymes, and antituberculous antibody serum levels have been proposed as possible markers to monitor disease activity in patients with tuberculosis. They have all shown limited sensitivity or specificity. The authors therefore attempted to generate a canonical variable using discriminant analysis, including sensitive and specific parameters, to be a reliable marker in classifying patients correctly during the course of pulmonary tuberculosis. The following parameters were selected: two soluble cell activation markers (soluble interleukin-2 receptor and sCD8); the levels of immunoglobulin (Ig) G and IgM antibodies against the A60 antigen complex; and the presence of specific antibodies directed to eight different A60 components, revealed by Western blot analysis. The tests were performed on sera from three groups of patients with pulmonary tuberculosis. The first group comprised 25 patients with onset tuberculosis, clinically active (OTCA), evaluated at the time of admission. The second group included 28 patients with chemotherapy-treated tuberculosis, clinically active (CTCA), 2 months after therapy had begun. The third group included 20 patients with tuberculosis, nonclinically active (TNCA), who had had at least 1 year of effective therapy. The authors obtained an 80.9% rate of correct classification for the three groups and a rate of 100% when OTCA and TNCA were compared. The patients with CTCA were scarcely differentiated and tended to be distributed into the two other groups. To improve the separation between patients with CTCA and those with OTCA, a second canonical variable was generated with a 91.7% rate of correct classification, as compared with 71% obtained using the sCD8 as the best single variable. The mean values of the last canonical variable were statistically different (Mann-Whitney test, P = .049) when stratified for acid fast bacilli-positive or negative CTCA patients (microscopic detection). Three patients, followed during the entire course their disease, were, as expected, correctly positioned with respect to the subsequent disease phases.

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.

Mutual interactions between HIV-1 and cytokines in adherent cells during acute infection.

The production of cytokines by HIV-infected cells from adherent tissues as well as their effects on HIV replication in the same cells were investigated. CD4-transfected HeLa-T4-6c epithelial cells, CD4-positive normal lung fibroblasts and CD4-negative RD rhabdomyosarcoma cells were infected with HIV-1. All cultures were permissive for virus replication, which was completed within 48-72 h by Hela-T4-6c and RD cells and 2-3 weeks in normal fibroblasts. During the course of HIV replication, a series of cytokines (particularly IL-6 and TNF alpha) was produced and released in parallel to the peak of virus growth, in amounts varying with the cell system studied. Treatment of cultures with recombinant cytokines given at concentrations in the range of those induced by HIV-1 indicated that IL-6 and TNF alpha caused an increase of: i) CD4 expression, ii) HIV absorption to uninfected cells, and iii) release of infectious virions by infected cells. The fact that HIV-1 absorption and spread can be mediated by HIV-induced cytokines may be relevant in the pathogenesis of the in vivo disease, as it may constitute a possible self-enhancing model of HIV infection also in the solid tissues.

Interleukin-6 and tumour necrosis factor levels decrease in the suction blister fluids of psoriatic patients during effective therapy.

BACKGROUND: Involvement of various cytokines in psoriasis pathomechanisms has previously been reported. OBJECTIVE: To better define the relationship between the disease severity and interleukin-6 and tumour necrosis factor alpha skin levels, these two cytokines were measured in suction blister fluids obtained from involved and uninvolved skin areas of psoriatic patients treated with UVB, beta-methasone dipropionate and salicylic acid ointment. METHODS: Determinations were performed by ELISA in fluids obtained from 6 patients with the Kiistala method, every 1-2 weeks for at least 1 month. RESULTS: During the observation period, all the patients showed disease improvement (median PASI score declined from 13.4 to 3.9) and significant decreases in the cytokine levels in all samples. CONCLUSION: These results are in agreement with a functional involvement of these two cytokines in the pathogenesis of the disease and their possible use as follow-up markers.

Behaviour of several 'progression markers' during the HIV-Ab seroconversion period. Comparison with later stages.

Two acute phase reactants, four cytokines, five soluble factors and lymphocyte subpopulations have been simultaneously evaluated in 16 subjects before and closely after the HIV-Ab seroconversion time. The same variables have also been determined in 50 HIV-Ab-negative high risk subjects, in 36 CDC II-III and in 30 CDC IV patients, utilizing a mixed longitudinal epidemiological model. The results show significant variations of few parameters in the early phases (increase: sCD8, beta-2-Microglobulin, sIL-2R, sCD23, Neopterin, IFN-alpha; decrease: CD4+ lymphocytes). In the course of the disease, many others parameters progressively increase (IFN-tau, IL-4, IL-6, acid-alpha 1-glycoprotein, alpha 1-antitrypsin) or decrease (B- and T-lymphocytes). Ferritin, in particular, highly increases only in CDC IV stage. These data may be useful to monitor patients during the entire course of their disease and to suggest the time elapsed from seroconversion.