Plant allergies are a frequent cause of visits to the allergologist. In the Alergologica 2005 study carried out in Spain, fruits were the main cause of food allergies [1]. Plant allergies in a specific population are related to consumption habits and to the pollen pattern of the region under study. The objective of this retrospective study was to determine the proteins responsible for fruit allergies in Asturias, a region situated on the Cantabrian Sea with an Atlantic climate and a pollen pattern different to that of the rest of Spain. We recruited patients diagnosed with fruit allergy from a database of patients seen in an outpatient clinic between 2010 and 2013. To be included in the study, all patients had to present a clinical picture characteristic of IgE-mediated allergy to fruits, with a positive prick-prick test or skin prick test result (ALKAbelló) with the culprit fruit. Sensitization to panallergens such as profilin, lipid transfer protein (LTP), and birch extract, as well as to Lolium extract was determined by skin prick test (ALK-Abelló), ImmunoCAP (Phadia) (rPhl p 12, rPru p 3, and nBet v 1), and ISAC 112 (Thermo Fisher Scientific). In vitro techniques were only used when the panallergen was not identified using the skin tests. The ImmunoCAP test was ordered for 10 patients and the ISAC test for 15. The study sample comprised 56 patients (29 men) with a median age of 29 years (IQR, 20-40). Symptoms triggered by Rosaceae fruits were detected in 22 patients (39%), kiwi in 16 patients (29%), peach only in 12 patients (21%), melon in 6 patients (11%), and banana in 4 patients (7%). Less frequent reactions were observed for fig, apple only, and orange (2 cases each, 4%) and avocado, strawberry, cherry, and passion fruit (1 case each, 2%). As for clinical manifestations, 63% of patients presented oral allergy syndrome (OAS), 16% urticaria/angioedema, 12% anaphylaxis, 5% contact urticaria, and 4% digestive symptoms. The proteins responsible for the reactions were LTPs (17 cases [30%]), pathogenesis-related (PR) protein 10 (17 cases [30%]), profilins (6 cases [11%]), Act d 1 (5 cases [9%]), unknown proteins (5 cases [9%]), PR-10 and profilins (4 cases [7%]), thaumatins (1 case [2%]), and chitinasaes (1 cases [2%]). The relationship between the type of fruit and the sensitization identified is shown in the Table. Only one-third of patients who were allergic to PR-10 had symptoms caused by allergy to Betulaceae pollen. However, 90% of those who were allergic to profilins had symptoms caused by allergy to grasses. Most cases of fruit allergy were caused by members of the Rosaceae family. PR-10 were the triggering proteins in 70% of cases. In a recent review of allergy to Rosaceae in Europe, PR-10 was shown to be of minor importance in Spain, as levels of sensitization were very low (0%-13%) [2]. Our findings contrast sharply with this result and are more consistent with findings from central Europe and Scandinavia, although in these regions, allergy to Rosaceae is significantly associated with allergy to pollen from Betulaceae, while in our region this was the case with only one-third of patients. Sensitization to profilin in patients who are allergic to Rosaceae in Spain has been reported to range from 56% to 80% [2]. In our study, levels were markedly lower (23%). Kiwi was the second most common cause of allergy, representing almost one-third of all cases. Act d 1 was the most frequently involved protein (one-third of cases). Curiously, this figure is similar to that found for Iceland in a recent European study on allergy to kiwi [3]. The proteins PR-10, LTP, and profilin all accounted for a similar percentage of cases. These Table. Culprit Allergens From the Fruits Involved in the Reactions

Allergic skin disorders include urticaria, angioedema, contact dermatitis and atopic dermatitis, but the model fitting most closely the systemic concept of allergy is atopic dermatitis (AD), the pathogenesis of which is linked to a complex interaction between skin barrier dysfunction and environmental factors such as allergens and microbes. In particular, an important advance was the demonstration that the mutation of the skin barrier protein filaggrin is related strictly to allergen sensitization and to the development of asthma in subjects with AD. The altered skin barrier function, caused by several factors, results in the passage of allergens through the skin and to systemic responses. A pivotal role in such a response is exerted by Langerhans cells which, via their immunoglobulin E (IgE) receptor, capture the allergens and present them to T cells. When T helper type 2 (Th2) cells are activated, the production of a proinflammatory cytokines and chemokines pattern sustains the persistence of inflammation. Known AD-related cytokines are interleukin (IL)-5, IL-13 and tumour necrosis factor (TNF)-alpha, with emerging importance for IL-17, which seems to drive airway inflammation following cutaneous exposure to antigens, and IL-31, which is expressed primarily in skin-homing Th2 cells. Skin-homing is another crucial event in AD, mediated by the cutaneous lymphocyte-associated antigens (CLA) receptor, which characterizes T cell subpopulations with different roles in AD and asthma.

RATIONALE: Rhinitis caused by sensitization to cypress pollen is generally considered particularly disturbing. A number of studies demonstrated the efficacy of allergen specific immunotherapy with cypress extracts, but thus far the effects of this treatment on quality of life are uninvestigated.METHODS: We evaluated the changes in quality of life (QoL) measured by the specific questionnaire RQLQ by E. Juniper in 34 subjects (20 males and 14 females, mean age 34 years, range 19-51 years) with cypress-induced rhinitis undergoing placebo-controlled sublingual immunotherapy (SLIT) with a high dose extract of Juniperus ashei from Stallergenes standardized in IR (index of reactivity). SLIT was performed by a build-up phase in 12 days and a maintenance phase of 300 IR/day up to the end of pollen season. RQLQ was submitted during the peak of cypress pollen season before and after the treatment. The data were analysed by non parametric tests.RESULTS: Nineteen patients received active treatment and 15 received placebo. QoL after SLIT significantly improved compared to baseline (Wilcoxon rank test, p = 0.038) in actively but not in placebo treated patients, and a significant difference was found in the QoL score in favour of actively treated patients compared to placebo in the cypress pollen season following SLIT (Mann Whitney U test, p = 0.019).CONCLUSIONS: SLIT with high doses of a standardized extract of Juniperus ashei is able to significantly improve the quality of life in subjects with seasonal rhinitis caused by cypress pollen. RATIONALE: Rhinitis caused by sensitization to cypress pollen is generally considered particularly disturbing. A number of studies demonstrated the efficacy of allergen specific immunotherapy with cypress extracts, but thus far the effects of this treatment on quality of life are uninvestigated. METHODS: We evaluated the changes in quality of life (QoL) measured by the specific questionnaire RQLQ by E. Juniper in 34 subjects (20 males and 14 females, mean age 34 years, range 19-51 years) with cypress-induced rhinitis undergoing placebo-controlled sublingual immunotherapy (SLIT) with a high dose extract of Juniperus ashei from Stallergenes standardized in IR (index of reactivity). SLIT was performed by a build-up phase in 12 days and a maintenance phase of 300 IR/day up to the end of pollen season. RQLQ was submitted during the peak of cypress pollen season before and after the treatment. The data were analysed by non parametric tests. RESULTS: Nineteen patients received active treatment and 15 received placebo. QoL after SLIT significantly improved compared to baseline (Wilcoxon rank test, p = 0.038) in actively but not in placebo treated patients, and a significant difference was found in the QoL score in favour of actively treated patients compared to placebo in the cypress pollen season following SLIT (Mann Whitney U test, p = 0.019). CONCLUSIONS: SLIT with high doses of a standardized extract of Juniperus ashei is able to significantly improve the quality of life in subjects with seasonal rhinitis caused by cypress pollen.

SummaryThe efficacy of specific immunotherapy and particularly of sublingual immunotherapy (SLIT) in subjects allergic to Cupressaceae pollen is well defined, but no study assessed its effects on quality of life (QoL). We evaluated the effectiveness of SLIT with a standardized, high‐dose extract of Juniperus ashei in patients with cypress‐induced rhinitis, using QoL as the major outcome measure. Thirty‐four patients, 20 males and 14 females, mean age 33.8 years, with allergic rhinitis (AR) from cypress pollen were randomly assigned to receive an allergen extract standardized in index of reactivity (IR) of J. ashei (19 patients) or a placebo (15 patients). The schedule was pre‐coseasonal, with a build‐up phase in 12 days and a maintenance treatment with 300 IR a day up to the end of the cypress pollen season. All patients registered in diary cards their symptoms, drug consumption and side‐effects. The QoL was measured before and after SLIT by the Rhinoconjunctivitis Quality of Life Questionnaire (RQLQ) by Juniper during the peak of the cypress pollen season. QoL significantly improved in respect to baseline in actively (P=0.017) but not in placebo‐treated patients, and there was a significant difference in favour of actively treated patients during the pollen season (P=0.02). Actively and placebo‐treated patients had comparable symptom–medication scores in the period from 15 February to 15 March, corresponding to the peak pollen season, but placebo‐treated patients showed an uneven drug consumption compared with actively treated patients. Side‐effects were relatively common, but mostly with local short‐lasting symptoms, and no systemic reaction was reported. These findings show that SLIT with high doses of J. ashei in subjects with cypress‐induced rhinitis significantly improves QoL and confirm previous observations on the disagreement between QoL – which assesses patient's perception – and medical parameters used in trials.

SUMMARY Since in coeliac disease mucosal flattening has been suggested to result from an increased enterocyte apoptosis triggered by Fas/Fas ligand system and perforin cytolytic granules, we looked for a similar mechanism in autoimmune enteropathy. Moreover, we tried to assess whether enterocyte autoantibodies, which are the hallmark of autoimmune enteropathy, may be involved in triggering enterocyte apoptosis in this condition. Immunohistochemical staining with anti-Fas, -FasL and -perforin MoAb, and TUNEL technique were applied on endoscopic duodenal biopsies of two autoimmune enteropathy patients, two untreated coeliac patients and two biopsied controls. Cytotoxicity assays were carried out by incubating peripheral blood mononuclear cells from a healthy subject (effectors) with enterocytes primed with patient or control sera (targets). In autoimmune enteropathy a large number of enterocytes were apoptotic, as in coeliac disease, whereas neither Fas/Fas ligand or perforin expressions were up-regulated. On the other hand, antibody-dependent cellular cytotoxicity assay revealed the ability of sera from patients with autoimmune enteropathy to mediate enterocyte death through apoptosis. These results point to enterocyte autoantibody-dependent cellular cytotoxicity as the prevalent mechanism of increased enterocyte apoptosis in autoimmune enteropathy but not in coeliac disease.

OBJECTIVE:The efficacy of probiotic organisms in the treatment of pouchitis has been reported. In the present study, we evaluated the tissue levels of pro- and anti-inflammatory cytokines, nitric oxide synthase, and matrix metalloproteinases in control and inflamed pouches before and after antibiotic and probiotic treatment of patients with acute pouchitis.METHODS:Pouch biopsy samples were obtained from seven patients with pouchitis before and after antibiotic and probiotic treatment. Tissue samples from five patients with normal pouches were used as controls. Cytokines were determined by ELISA, matrix metalloproteinase activity was evaluated by zymograms, and nitric oxide synthase activity was determined by measuring arginine to citrulline conversion.RESULTS:Tissue levels of tumor necrosis factor α increased (p 0.01) in pouchitis relative to uninflamed pouches and reduced after antibiotic and probiotic treatment. Also, interferon γ and interleukin 1α (IL-1α) augmented in pouchitis, but their increase did not reach statistical significance. The latter, however, were lower (p < 0.05) after treatment with the antibiotics and probiotics. Tissue levels of IL-4 and IL-10 were unchanged in inflamed pouches and unaffected by antibiotic treatment. However, IL-10 increased (p < 0.05) after probiotic treatment. Moreover, inflamed pouches had higher levels of inducible nitric oxide synthase and gelatinase activities, which decreased after treatment.CONCLUSIONS:The ability of antibiotic and probiotic treatments to increase tissue levels of IL-10, at a higher level than those observed in control pouches, and to decrease, to levels present in control pouches, proinflammatory cytokine, inducible nitric oxide synthase, and matrix metalloproteinase activity may suggest a mechanism of action to explain the efficacy of this therapeutic regime in pouchitis.

Systemic sclerosis (SSc) is a connective tissue disease in which immune system activation is evidenced by high levels of different cytokines in the sera and/or in the supernatants of cultured peripheral blood mononuclear cells (PBMC) and by the presence of specific autoantibodies. gamma/delta T cells accumulate in the lung and the skin of SSc patients suggesting their potential role in the development and maintenance of the disease. The aim of this study was to assess cytokine production and cytotoxic activity of circulating gamma/delta T lymphocytes obtained from SSc patients and to evaluate their potential role during this disorder. Our results showed that both the proportion and the absolute number of IFN-gamma gamma/delta-producing cells (i.e. displaying a Th1 polarization) in SSc was significantly higher than either the proportion and the absolute number of IL-4 gamma/delta-producing cells in SSc or the proportion and the absolute number of IFN-gamma gamma/delta-producing cells in healthy controls (P < 0.05 for both groups). Furthermore, the cytotoxic activity of enriched gamma/delta T cells was significantly increased in SSc patients compared with controls. The results concerning the Vdelta1+ T cell subset paralleled those of total gamma/delta T lymphocytes. In contrast, alpha/beta T cells from SSc and control subjects displayed Th2 cytokine production. All these findings were independent of both disease subset and clinical status. Our data demonstrate that, although SSc is generally considered a Th2 autoimmune disease, Th1 polarization of gamma/delta T cells and an increase in their cytotoxic activity is observed in SSc, suggesting that gamma/delta T cells could have a relatively autonomous role in the pathogenesis in this disease.

Background-Lamina propria (LPLs) and intraepithelial (IELs) lymphocytes are markedly increased in coeliac mucosa, and are thought to play a crucial role in the generation of villous atrophy in coeliac disease (CD). However, the mechanisms by which they mediate the killing of enterocytes in this condition are still poorly characterised.Aim-We investigated Fas mediated cytotoxicity and apoptosis of both LPLs and IELs, isolated from 10 untreated coeliac patients, 10 coeliac patients on a gluten free diet, and 10 biopsied controls.Methods-Fas and Fas ligand expression were assessed by flow cytometry and immunocytochemistry. Lymphocyte cytotoxicity against Fas expressing Jurkat cells was determined by the jam test. The effect of the antagonist ZB4 anti-Fas antibody on apoptotic activity exerted by coeliac lymphocytes against enterocytes was analysed. Lymphocyte apoptosis was assessed by oligonucleosome ELISA.Results-LPLs and IELs showed increased apoptotic activity and higher levels of Fas ligand expression in untreated CD compared with treated CD patients and controls. Enterocyte apoptosis observed after coculturing coeliac lymphocytes and enterocytes in the presence of ZB4 antibody was reduced. In active CD, LPLs manifested increased apoptosis whereas IELs showed decreased apoptosis.Conclusions-Our results support the involvement of the Fas/Fas ligand system in CD associated enterocyte apoptosis. Increased LPL apoptosis is likely to down-regulate mucosal inflammation whereas decreased IEL apoptosis could be responsible for autoimmune and malignant complications of CD.

In coeliac disease (CD) immunological abnormalities are not confined to the small bowel and it has been suggested that changes in peripheral blood lymphocytes (PBL), such as lymphopenia and increased T-cell activation, may predispose to malignant or autoimmune complications of this condition. In the light of the recent findings about the Fas-Fas ligand (FasL) system in regulating lymphocyte homeostasis, the aim of the present study was to investigate peripheral lymphocyte Fas-mediated apoptosis in CD to establish whether the homeostatic role of apoptosis in peripheral T-cell selection is maintained. Moreover, because a soluble form of Fas has been described to be functionally implicated in the Fas signalling system, suggesting a relationship between some disorders and soluble Fas function, we measured levels of soluble Fas in sera of coeliac patients and analysed the relationship between these levels and the proportions of apoptotic and Fas(+) PBL to further explore the function of the Fas-FasL pathway in this condition. Finally, we evaluated whether the increased prevalence of anticardiolipin antibodies, recently described in CD, could be related to PBL apoptosis in this condition. We demonstrated an increased apoptosis and higher levels of Fas and FasL expression in PBL isolated from untreated coeliac patients when compared to treated coeliac patients and controls. In addition, low levels of soluble Fas and a significant positive correlation between anticardiolipin antibodies and PBL apoptosis were found in untreated CD. Then, our results showed an increased susceptibility of PBL to undergo Fas-mediated apoptosis in active CD. This increased apoptosis could be responsible for both lymphopenia and immunogenic exposure of phospholipids with subsequent production of autoantibodies.

Our aim was to evaluate whether increased enterocyte apoptosis was responsible for mucosal flattening in celiac disease (CD), and, since the mechanisms responsible for tissue injury in this condition are unknown, we studied the possibility that the Fas-Fas ligand (FasL) system may be involved. Endoscopic duodenal biopsy specimens from 12 patients with untreated and 12 with treated CD and 12 control subjects were evaluated for enterocyte apoptosis by the terminal deoxynucleotidyl transferase-mediated digoxigenin-deoxyuridine triphosphate nick-end labeling assay and for Fas and FasL expression by immunohistochemistry. A coculture of isolated enterocytes (targets) and purified lamina propria mononuclear cells (LPMCs) (effectors) was performed in the absence or presence of an antagonistic ZB4 anti-Fas antibody. We found a significant correlation between the degree of villous atrophy, morphometrically evaluated, and the level of enterocyte apoptosis, suggesting that mucosal flattening is a consequence of exaggerated epithelial cell death. Most celiac enterocytes express Fas, and LPMCs express FasL. The abolishment of enterocyte apoptosis observed in the presence of ZB4 antibody suggests that enterocytes are potential targets of lymphocyte infiltrate. These results directly demonstrate that FasL-mediated apoptosis is a major mechanism responsible for enterocyte death in CD.

The aim of the present work was, first, to analyze the apoptotic effect in vitro of sonicated preparations of selected strains of lactic acid bacteria on normal and tumor human lymphocytes. Incubation with bacterial samples led to a relevant time-dependent apoptotic cell death of Jurkat cells but not normal human peripheral blood lymphocytes. Lactobacillus brevis (CD2) samples were more efficient in inducing apoptosis of Jurkat cells than were samples of Streptococcus thermophilus (S244). In an attempt to characterize the mechanisms underlying these effects, we found that the apoptotic death-inducing ability of S244 preparations could be attributed to the ability of high levels of neutral sphingomyelinase activity to generate relevant amounts of ceramide, a known apoptotic death messenger, in Jurkat cells. On the other hand, our results indicate that apoptosis induced by CD2 samples could also be associated with high levels of arginine deiminase activity, which in turn was able to downregulate polyamine synthesis in Jurkat cells.