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Abundant heterogeneous immune cells infiltrate lesions in chronic inflammatory diseases and characterization of these cells is needed to distinguish disease-promoting from bystander immune cells. Here, we investigate the landscape of non-communicable inflammatory skin diseases (ncISD) by spatial transcriptomics resulting in a large repository of 62,000 spatially defined human cutaneous transcriptomes from 31 patients. Despite the expected immune cell infiltration, we observe rather low numbers of pathogenic disease promoting cytokine transcripts (IFNG, IL13 and IL17A), i.e. >125 times less compared to the mean expression of all other genes over lesional skin sections. Nevertheless, cytokine expression is limited to lesional skin and presented in a disease-specific pattern. Leveraging a density-based spatial clustering method, we identify specific responder gene signatures in direct proximity of cytokines, and confirm that detected cytokine transcripts initiate amplification cascades of up to thousands of specific responder transcripts forming localized epidermal clusters. Thus, within the abundant and heterogeneous infiltrates of ncISD, only a low number of cytokine transcripts and their translated proteins promote disease by initiating an inflammatory amplification cascade in their local microenvironment. Inflammatory skin diseases involve various different immune cells in a localised area. Here the authors use spatial transcriptomics to show that disease relevant cytokine transcripts are sparsely expressed in lesional skin, yet are associated with local amplification cascades that promote skin inflammation. [ABSTRACT FROM AUTHOR]

Background: To our knowledge, an international consensus is lacking regarding the development of an adequate informed consent form for a patch test (PT) and the information that should be included in such document. Objectives: The aim of the study was to reach a consensus on the specific points that need to be addressed in a PT consent form. Methods: A Delphi survey, comprising 2 rounds and 1 final discussion, was used to gather and analyse data, which was conducted over the Internet. Each statement that reached a consensus with the respondents (9 expert dermatologists from Europe) was defined as a median consensus score (MED) of ≥7 and agreement among panelists as an interquartile range (IQR) of ≤3. All study participants were members of the EADV task force on contact dermatitis. Results: The expert panel addressed several topics that should be included in an informed consent form for a PT: introduction, preparation for PT, testing procedure, allowed activities, adverse events and additional authorizations. Conclusions: Our results assess recommendations regarding points to be contained in an informed consent form for a PR. Future actions towards standardization and harmonization of this specific consent form are needed. [ABSTRACT FROM AUTHOR]

Summary: Background: Interleukin (IL)‐26 is a signature T helper 17 cytokine described as a proinflammatory and antimicrobial mediator. So far, IL‐26 has been reported in several immune‐mediated inflammatory diseases, but its involvement in inflammatory skin disorders is poorly known. Objectives: To investigate the role of IL‐26 in hidradenitis suppurativa (HS), through its involvement in antimicrobial activity. Methods: IL‐26 was assessed in patients with HS through gene expression and protein analysis at skin and circulating levels. Ex vivo HS organ skin cultures, together with IL‐26 antibody treatment, were performed to determine the IL‐26 activity. Peripheral blood mononuclear cells (PBMCs) from patients with HS and healthy controls were either silenced or not with IL‐26 small interfering (si)RNA in order to measure its antimicrobial, cytotoxic and phagocytic activities against Staphylococcus aureus. Results: Firstly, we observed that IL‐26 is able to modulate the proinflammatory response at the immune cell level. IL‐26 was increased in the plasma of patients with HS compared with healthy controls. Subsequently, we explored the bactericidal, cytotoxic and phagocytic activities of PBMCs against S. aureus in patients with HS and healthy controls. These activities were lower in patients with HS than in controls. Remarkably, the killing activities were reduced when healthy control PBMCs were transfected with IL‐26 siRNA. However, the transfection did not affect the killing activity of HS PBMCs, supporting the idea that IL‐26 lacks efficacy in HS. Conclusions: Our findings suggest that infection susceptibility in HS might be related to IL‐26. Although the role of bacteria remains controversial in HS, this paper supports that there is a defect of antimicrobial response in these patients. What's already known about this topic? Interleukin (IL)‐26 is a T helper 17 cytokine described as an antimicrobial and proinflammatory mediator.IL‐26 has been reported in immune‐mediated inflammatory diseases, but its involvement in inflammatory skin disorders remains unclear.Hidradenitis suppurativa (HS) is a chronic inflammatory skin disorder characterized by deficiency of IL‐20 and IL‐22 (a close homologue of IL‐26), which causes antimicrobial peptide pauperization leading to severe and recurrent skin infections. What does this study add? IL‐26 plasma levels are higher in patients with HS than in healthy control individuals.The antimicrobial activity of IL‐26 might be ineffective in patients with HS. What is the translational message? Cutaneous antimicrobial incompetence in HS could be related to IL‐26. Linked Comment: Giamarellos-Bourboulis. Br J Dermatol 2019; 181:892–893. Respond to this article [ABSTRACT FROM AUTHOR]

Dear Editor, Atopic dermatitis (AD) and psoriasis (Pso) are chronic diseases that may be phenotypes at polar ends of the T-cell inflammatory milieu.1,2 The two diseases represent two distinct pathogenetic entities, with T-helper (Th) type 2 (Th2) cytokines interleukin (IL)-4, IL-13, IL-31 and possibly IL-22 mainly implicated in AD, and Th17 cytokines IL-17 and IL-23 in Pso.3 However, a range of phenotypes with overlapping clinical patterns exist, possibly reflecting a variable expression of T-cell axes.1 Dupilumab is a human monoclonal antibody directed against the subunit of the IL-4 receptor, approved for the treatment of moderate-to-severe AD. Patients with D-PsoR and patients with AD had suffered from persistent AD since childhood, the mean age at symptom onset being 1-8 ± 5-2 vs. 2-6 ± 4-9 years, respectively. We observed a strong increase of IL-23A level in D-PsoR with respect to both Pso and AD lesional skin, whereas the IL-23A levels in D-PsoR nonlesional skin were decreased with respect to AD nonlesional skin. [Extracted from the article]

Background: Given the limited alleviation of chronic pain with pharmacological treatments, various nonpharmacological and self-care approaches are often proposed that require patients' motivation.Objective: To evaluate the level of readiness (LOR) to practise different types of active self-care among chronic pain patients.Method: A quantitative cross-sectional survey was conducted among all chronic pain patients seeking care at the Pain Center of an academic hospital from June 2013 to March 2015. Sociodemographic data, pain characteristics, treatments and the LOR to practise active self-care were investigated.Results: Among the 1524 eligible patients, 639 (41.9%) were included. The median pain duration was 8.5 years (interquartile range = 7.5). Two-thirds (63.7%) of the patients reported high pain-related disability, and 64.6% had used opioids during the previous six months. Most patients had a high (44.1%) or moderate (24.6%) LOR to practise active self-care. Multivariable multinomial regression analysis showed that independent factors associated with a high LOR were a higher level of education (relative risk ratio (RRR) = 3.42, 95% confidence interval (CI): 1.90-6.13, p < 0.001), unemployed status due to medical condition (RRR = 2.92, 95% CI: 1.30-6.56, p = 0.009), the use of dietary supplements 'against pain' (RRR = 2.77, 95% CI: 1.52-5.04, p = 0.001) and neuropathic pain characteristics (RRR = 1.80, 95% CI: 1.40-3.12, p = 0.036). Older age was a factor predicting a lower LOR (RRR = 0.97, 95% CI: 0.94-0.99, p = 0.039). Long-term chronic pain, severe pain-related disability and the presence of a mood disorder were not associated with a lower LOR.Conclusion: Most chronic pain patients, including those severely affected, indicated their readiness to practise active self-care methods.Significance: Most chronic pain patients, even those severely affected, appeared to be ready to practise active self-care therapies and we believe that further studies are needed to investigate their impact on pain and quality of life. [ABSTRACT FROM AUTHOR]

Nine hundred and thirty-nine rPla a 1, nPla a 2, and rPla a 3 Immuno CAP ISAC reactors were studied. nPla a 2pos MUXF3pos but Pla a 1/2neg subjects were excluded from the study because they were cross-reactive carbohydrate determinant reactors. Among the 764 remaining participants, 71.9% were Pla a 3pos, 54.1% Pla a 2pos, and 10.9% Pla a 1pos. Among Pla a 3 reactors, 89.6% were Pru p 3pos and 86.8% Jug 3pos, but the strongest IgE recognition relationship was observed between Pla a 3 and Jug r 3. Distinctive clinical subsets could be documented among plane tree-allergic patients. Pla a 3 reactors had both local and systemic food-induced reactions, but lower past respiratory symptoms occurrence. Pla a 2 reactivity was associated with respiratory symptoms but inversely related to systemic reactions to food. Cosensitization to Pla a 2 and Pla a 3 was associated with a lower past incidence of severe food-induced reactions. [ABSTRACT FROM AUTHOR]

The article focuses on a study on influence of profilin hypersensitivity on the clinical expression of allergic disease. It mentions profilin sensitization may underlie severe food-induced allergic reactions in some patients but also exert a protective effect against systemic reactions associated with the sensitization to a stable allergen such as nsLTP. It also mentions adverse reactions to foods, including oral allergic syndrome and urticaria.

Background Nonspecific lipid transfer proteins (ns LTPs) represent a major cause of systemic food allergic reactions in the Mediterranean area. This study investigate hierarchical patterns and cluster relationships of IgE sensitization to different ns LTPs, and the relationship to clinical allergy in a large Italian cohort. Methods A total of 568 ns LTP-positive subjects after IgE Immuno CAP- ISAC microarray analysis with Ara h 9, Art v 3, Cor a 8, Jug r 3, Pla a 3, Pru p 3 and Tri a 14 allergens were studied. IgE inhibition experiments were carried out with mugwort and plane tree pollen extracts. Results Eighty-two per cent of ns LTP-positive participants (94% if <6 years old) were Pru p 3pos, and 71% were Jug r 3pos. Participants who reacted to >5 ns LTPs reported a higher incidence of food-induced systemic reactions. Only Art v 3 and Pla a 3 (mugwort and plane tree ns LTPs, respectively) were associated with respiratory symptoms, and a correlation was observed between sensitization to pollen and plant food ns LTPs, particularly between Pla a 3 and tree nut/peanut ns LTPs. Co-sensitization to Par j 2 and PR-10 or profilin pan-allergens was associated with a lower prior prevalence of severe food-induced reactions. In inhibition assays, plane and mugwort pollen extracts inhibited 50-100% of IgE binding to food ns LTPs in microarrays. Conclusions Testing IgE reactivity to a panel of ns LTP allergens unveils important associations between ns LTP sensitization profiles and clinical presentation and allows the identification of novel cluster patterns indicating likely cross-reactivities and highlighting potential allergens for ns LTP immunotherapy. [ABSTRACT FROM AUTHOR]

Background Specific food-dependent exercise-induced anaphylaxis ( S- FDEIAn) is a distinct form of food allergy in which symptoms are elicited by exercise performed after ingesting food to which the patient has become sensitised. Non-specific FDEIAn ( NS- FDEIAn) is a syndrome provoked by exercise performed after ingesting any food. Objective We sought to identify the culprit allergenic molecules in patients with FDEIAn, combining 'classic' allergy testing with an allergenic molecule-based microarray approach for Ig E detection. Methods All subjects were evaluated who reported at least one episode of anaphylaxis in association with physical exercise performed within 4 h after a meal. We performed skin prick tests ( SPT) with commercial food extracts, prick plus prick tests ( P + P) with fresh foods ( P + P), and serum specific Ig E assays by means of both the Immuno CAP ( CAP) and the ISAC 89 microarray system ( ISAC). Results Among our 82 FDEIAn patients, the most frequent suspected foods were tomato, cereals, and peanut. SPT, P + P, and CAP displayed different degrees of sensitivity. Each test disclosed some positivities not discovered by others. Seventy-nine subjects were positive to at least one food (49 to more than 20), whereas three were negative. All suspected foods were positive to at least one of SPT, P + P, and CAP. When tested using the ISAC, 64 (78%) subjects were positive to Pru p 3 [peach lipid transfer protein ( LTP)], 13 were positive to other food allergen molecules, and five displayed negative results to all food allergenic molecules. Overall, 79 patients probably had S- FDEIAn and the other 3 NS- FDEIAn. Conclusions Multiple food hypersensitivity represents a clinical hallmark of a large percentage of FDEIAn patients. The very high prevalence of Ig E to the LTP suggests a role of this allergen group in causing S- FDEIAn. [ABSTRACT FROM AUTHOR]

Summary Background Egg allergy is a very common finding in early childhood. Detecting hen's egg ( HE) allergy outgrowing and reintroduction of food containing egg is a task for the allergist. Objective We sought to evaluate the suitability of boiled egg food challenge compared with IgE to allergenic molecules from HE white using a microarray system. Method Sixty-eight children referring to our centre by the family paediatricians for a suspected egg allergy were enrolled. Patients underwent double-blind, placebo-controlled food challenge with boiled and raw eggs. Challenge outcomes were compared with skin tests performed using egg white and yolk commercial extracts, to prick-prick test with boiled and raw egg white and yolk, total IgE, egg white specific IgE detected using Immuno CAP and IgE to egg allergens available on the immunosolid phase allergen chip ( ISAC) 103 microarray. Result Nineteen subjects (28%) were reactive to both raw and boiled egg, 14 (20.5%) to raw egg only and 35 (51.4%) tolerated both boiled and raw egg. Efficiency analysis was carried out using both raw and boiled egg challenges as gold standard. Forty four of 47 Gal d 1 negative patients tolerated boiled egg (94%). Conversely, 20 of 21 Gal d 1 positive patients reacted to raw egg (95%). None of the other tests was able to discriminate patients' response to HE challenge. Furthermore, Gal d 1 positivity seems to lead to broader environmental allergen IgE sensitization. Conclusion and Clinical Relevance The Gal d 1 IgE reactivity appears to be a very good predictor of HE clinical allergy. Gal d 1 positive children have a high frequency of HE allergy, whereas Gal d 1 negative children have a high frequency of tolerance to boiled egg. Multiple specific IgE detection by means of ISAC improves the diagnostic approach in HE allergic children, disclosing other food and inhalant allergic sensitizations, anyhow requiring a comprehensive clinical evaluation. [ABSTRACT FROM AUTHOR]

Summary Immune dysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX) is a rare syndrome due to a mutation in the forkhead box protein 3 gene ( FOXP3) leading to an impaired regulatory T cell (Treg) activity associated both with skewed T helper type 2 (Th2) response and autoreactive phenomena. The purpose of this study was to describe a combined proteomics and genomics approach to comprehensively evaluate clinical and immunological phenotypes of patients affected by IPEX. T cell receptor (TCR)-Vβ repertoire and peripheral blood lymphocytes phenotype from three brothers affected by IPEX were studied by flow cytometry. Specific immunoglobulin (Ig)E were evaluated by means of an allergenic molecules microarray [immuno solid-phase allergen chip (ISAC)]. Total RNA was extracted and hybridized to Affymetrix oligonucleotide arrays to obtain quantitative gene-expression levels. No FOXP3 protein was detectable within CD127-CD25highCD4+ T cells from peripheral blood. A T cell-naive phenotype (CD62L+CD45R0-) associated with a reduction of both CD26 and CD7 expression and a TCR-Vβ 8 and 22 family expansions were found. B lymphocytes were mainly CD5+ (B1) cells expressing a naive phenotype (tcl1+CD27-). The three IPEX patients had severe food allergy and specific IgE reactivity to cow's milk allergens, a hen's egg allergen and a wheat allergen. Gene expression profile analysis revealed a dysregulation associated mainly with Th1/Th2 pathways. The multiplexing evaluation reported in this study represents a comprehensive approach in the assessment of genetic conditions affecting the immune system such as the IPEX syndrome, paving the way for the development of diagnostic tools to improve the standard clinical and immunological profiling of the disease. [ABSTRACT FROM AUTHOR]

Background: Nonspecific lipid transfer proteins (nsLTPs) are important food allergens. Often, patients allergic to the nsLTP in peach suffer from allergy to hazelnuts. We aimed to analyse the T-cell response to Cor a 8, the nsLTP in hazelnut and its immunological cross-reactivity with the nsLTP in peach, Pru p 3. Methods: Cor a 8-reactive T-cell lines (TCL) established from patients allergic to hazelnut and peach were stimulated with 12-mer peptides representing the complete amino acid sequence of Cor a 8 to identify its T-cell-activating regions and with Pru p 3 to investigate cellular cross-reactivity. T-cell clones specific for different major T-cell-activating regions of Pru p 3 were stimulated with Cor a 8. Both nsLTPs were subjected to endolysosomal degradation assays. Immunoglobulin E (IgE) cross-reactivity between Cor a 8 and Pru p 3 was assessed in inhibition enzyme-linked immunosorbent assay. Results: No major T-cell-activating region was found among 26 T-cell-reactive peptides identified in Cor a 8. Although generated with Cor a 8, 62% of the TCL responded more strongly to Pru p 3. This cross-reactivity was mediated by T cells specific for the immunodominant region Pru p 361-75. Peptide clusters encompassing this region were generated during lysosomal degradation of both nsLTPs. Cor a 8 was more rapidly degraded by lysosomal proteases than Pru p 3. Pre-incubation of sera with Pru p 3 completely abolished IgE binding to Cor a 8, which was not the case vice versa. Conclusions: T-cell reactivity to Cor a 8 is predominantly based on cross-reactivity with Pru p 3, indicating that the latter initiates sensitisation to its homolog in hazelnut. The limited allergenic potential of Cor a 8 seems to be associated with rapid lysosomal degradation during allergen processing and the lack of major T-cellactivating regions. [ABSTRACT FROM AUTHOR]

Background The availability of allergenic molecules and high-throughput microtechnologies allow the collection of a large number of IgE results at the same time in a single test. This can be carried out applying the test in the routine diagnostic work-up. Objective The aim of this study was to make a cross-sectional evaluation of the raw prevalence of IgE reactivity to allergenic molecules in serum samples from a cohort of Italian patients using an innovative tool. Methods The ISAC, a microarray system, has been used for specific IgE detection using 75 different allergenic molecules. Sera were collected from 23 077 unselected consecutive individuals complaining about any allergic disease. Results Sixteen thousand four hundred and eight of 23 077 patients had IgE to at least one of 75 allergenic molecules. The top-ranked molecules in this cohort were Cup a 1 (42.7%), Der f 2 (38.7%), and Phl p 1 (37.9%), whereas all the other allergens tested scored in a range between 36.8% and 0.04%, including the first food allergen, Pru p 3, ranked 15th (9.79%). Prevalence varied quite markedly depending on the age range considered, and showing a different behaviour in the lifetime sensitization process. Unsupervised two-way hierarchical clustering analysis generated distinctive patterns of reactivity as the result of IgE recognition of either homologous allergens belonging to different biological sources or non-homologous belonging to the same biological source. Conclusions Allergen-based microarray is a tool for the detection of IgE-related sensitization to panels of allergens and gives a more precise and comprehensive evaluation for an IgE-based epidemiology. This insight brings data for better understanding of the sensitization process. [ABSTRACT FROM AUTHOR]

Background: The clinical relevance of immunoglobulin E (IgE) to plant glycans is a longstanding debate. We sought to evaluate their clinical reactivity using the human glycoprotein lactoferrin expressed in rice. Methods: Allergic patients with IgE antibodies against plant glycans were analyzed for the presence of IgE against rice-produced lactoferrin. The potency of IgE to induce mediator release was assessed by basophil histamine release and skin prick tests (SPTs). Clinical relevance was evaluated by double-blind placebo-controlled oral challenge (DBPCOC). Results: Twenty-four of 29 sera (82.7%) with IgE antibodies against plant glycans demonstrated IgE binding to transgenic lactoferrin. In three of five cases transgenic lactoferrin induced histamine release. Compared to a control major grass pollen allergen lactoferrin concentrations needed for biological activity of IgE were 5–6 orders of magnitude higher. Skin prick test and DBPCOC were negative in five patients with potential clinical reactivity that volunteered to undergo these in vivo challenges. Conclusions: Poor or no biological activity and lack of clinical relevance of IgE-binding plant glycans (five out of five) was demonstrated using human lactoferrin expressed in rice as a model. [ABSTRACT FROM AUTHOR]

Mental retardation (MR) is a nonprogressive condition characterized by a significant impairment of intellectual capabilities with deficit of cognitive and adaptive functioning and onset before 18 years. Mental retardation occurs in about 2 to 3% of the general population and it is estimated that 25 to 35% of the cases may be due to genetic causes. Among these 'genetic' MR, 25 to 30% are probably due to mutations in a gene on the X chromosome (X-linked mental retardation, XLMR). Given the genetic heterogeneity of XLMR, the availability of a considerable number of patients with accurate phenotypic classification is a crucial factor for research. The X-linked Mental Retardation Italian Network, which has been active since 2003, has collected detailed clinical information and biological samples from a vast number of MR patients. Collected samples and clinical information are inserted within the XLMR bank, a comprehensive molecular and clinical web-based database available at the address . The database is organized in three distinct parts. Part I and II contain several electronic schedules to register information on the family, the phenotypic description, the photographs, and a 20 sec movie of the patient. Part III allows the registration of molecular analyses performed on each case; samples and clinical data are usable via password-restricted access. Clinical and molecular centers interested in joining the network may request a password by simply contacting the Medical Genetics of the University of Siena. The XLMR bank is an innovative biological database that allows the collection of molecular and clinical data, combines descriptive and iconographic resources, and represents a fundamental tool for researchers in the field of mental retardation. Hum Mutat 28(1), 13-18, 2007. © 2006 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

We studied the expression of adhesion molecules affecting recirculation and homing on peripheral blood CD4+ T cells of patients with systemic sclerosis (SSc), in order to evaluate whether the distribution of tissue targeted subsets could reflect the participation of internal organs or the extent of cutaneous involvement[i.e. limited cutaneous (lc) and diffuse cutaneous (dc)]. Peripheral blood mononuclear cells (PBMC) from 51 patients with SSc and 19 sex- and age-matched controls were investigated by cytofluorimetric analysis for lymphocyte subpopulations carrying the following surface molecules: CD3, CD4, CLA,α4β7 andα4β1. Standard routine biochemistry and clinical examinations were also performed in all patients. We found that bothα4β1+ andα4β7+ cells within the CD4+ T cell population were significantly increased, while CLA+ CD4+ T cells were significantly reduced in SSc, compared to healthy donors. Significantly lower absolute numbers ofα4β7+ cells were found in lc- compared to dc-SSc. Patients with oesophageal involvement had high numbers ofα4β7+ cells, while those with nephritis also showed low levels of CLA+ cells. Lung involvement was related directly toα4β1+ cell numbers and inversely toα4β7+ CD4 cell numbers. Taken together, our findings demonstrate that distinct CD4+ T cell populations with selective homing properties show changes from normal distribution in SSc, and such changes are related to clinical expression and organ involvement in the course of the disease. [ABSTRACT FROM AUTHOR]

Background: Rett syndrome is a severe neurodevelopmental disorder, almost exclusively affecting females and characterised by a wide spectrum of clinical manifestations. Bath the classic form and preserved speech variant of Rett syndrome are due to mutations in the MECP2 gene. Several other variants of Rett syndrome have been described. In 1985, Hanefeld described a variant with the early appearance of convulsions. In this variant, the normal perinatal period is soon followed by the appearance of seizures, usually infantile spasms. We have observed two patients with signs of Rett syndrome showing acquired microcephaly and stereotypic midline hand movements. The disease started with generalised convulsions and myoclonic fits at 1.5 months in the first patient and with spasms at 10 days in the other, suggesting a diagnosis of the Hanefeld variant. In these patients, MECP2 paint mutations and gross rearrangements were excluded by denaturing high performance liquid chromatography and real time quantitative PCR. The ARX and CDKL5 genes have been associated with West syndrome (infantile spasms, hypsarrhythmia, and mental retardation). Methods: Based on the clinical overlap between the Hanefeld variant and West syndrome, we analysed ARX and CDKL.5 in the two girls. Results: We found frameshift deletions in CDKL.5 in both patients; one in exon 5 (c. 163_166delGAAA) and the other in exon 18 (c.2635_2636delCT). CDKL5 was then analysed in 19 classic Rett and 15 preserved speech variant patients, all MECP2 negative, but no mutations were found. Conclusion: Our results show that CDKL5 is responsible far a rare variant of Rett syndrome characterised by early development of convulsions, usually of the spasm type. [ABSTRACT FROM AUTHOR]

Systemic sclerosis (SSc) is a connective tissue disorder characterized by excessive collagen deposition in the skin and internal organs. Several cytokines and chemokines have been implicated in the induction of fibrosis, but a definitive relationship between specific cytokines and organ involvement has not been established yet. Serum samples, PBMC and T cell lines (TCL) obtained from 54 patients affected by SSc and 20 healthy donors (HD) were examined by ELISA for Interferon-y (IFN-γ), interleukin (IL)-4, IL-6, IL-b, IL-18, Transforming growth factor (TGF)-Β1, Tumour necrosis factor (TNF)-α, sCD30, Macrophage derived chemokine (MDC), Monocyte chemo attractant protein (MCP) -1, Macrophage inflammatory protein (MIP)-1α and Regulated on activation normal T-cell expressed and secreted (RANTES). In all the SSc serum samples, we found significantly increased levels of IL6, TNFα and MCP-1 but reduced amounts of γ-IFN and MDC. IL6, IL10, IL18, MIP-1α and TNFα measured in supernatants from PHA-stimulated PBMC and 1L6, MCP- 1 and RANTES in supernatants from stimulated TCL were also increased in patients. MDC was decreased in all the biological SSc sources studied. TGF-Β1, IL10, and sCD30 were produced at a significantly lower level by SSc TCL. Serum IL6 and sCD30 levels were significantly increased in dc-SSc patients compared to lc-SSc as were levels of MCP-1 produced by PBMC and IL10 from TCL. We observed a strict relationship between pulmonary fibrosis and IL10, MCP-1 (both from TCL) and serum 1L6. Kidney involvement was related to serum MCP-i levels and IL18 production from PBMC. Oesophageal involvement correlated with MDC production from PBMC and IL10 synthesis by TCL. We showed that IL-6, IL-10, MDC and MCP-1 are variably associated with internal organ involvement and allow the discrimination between limited and diffuse forms of the disease. [ABSTRACT FROM AUTHOR]

Background: Multiple drug allergy syndrome is a clinical condition characterized by reactions against more than one different class of, both pharmacologically and structurally, unrelated drugs. Scanty data are available to date about a multiple drug delayed hypersensitivity syndrome. Our aim was to report the case of a delayed reaction to both β -methasone (β -MT) and penicillin-G (pen-G) occurring in the same patient, and analyse β -MT- and pen-G-specific T-cell Lines (TCLs) with regard to their specificity, phenotype and cytokine profile. Methods: We generated two drug-specific TCLs from biopsies at the site of positive intradermal reactions, and analysed their immunophenotype, T-cell receptor Vβ (TCR-Vβ ) domains expression and cytokine profile. Results: We demonstrated the specificity of the T cells isolated from positive intradermal test reactions to pen-G and β -MT through the strict dose-dependent proliferation in response to drug-pulsed autologous antigen presenting cells. Fluorescence activated cell sorter (FACS) analysis revealed a predominance of CD4+ cells in the inflammatory cell infiltrate of intradermal test with β -MT, while a predominance of CD8+ T cells in the site of delayed reaction to pen-G was found. The drug specific CD4+ and CD8+ T cells were heterogeneous, with regard to TCR-Vβ usage. CD8+ pen-G-TCL displayed a preferential T helper 2 (Th2) profile, while a substantially heterogeneous pattern of cytokine production characterized specific β -MT TCL. Conclusion: The study describes the coexistence in the same patient of a delayed hypersensitivity to both penicillin G and β -MT, driven, respectively, by pen-G-specificTh2-skewed CD8+ and β -MT specificTh0 CD4+ T cells. This case further support the existence of a multiple drug allergy syndrome also for delayed hypersensitivity. [ABSTRACT FROM AUTHOR]

Proinflammatory cytokines are deemed to play a pivotal role in the pathogenesis of multiple sclerosis (MS). They provide signals for T-cell activation and inflammatory cell recruitment in the brain and might directly alter neuroglial and neuronal cell survival and function. We found that peripheral blood mononuclear cells (PBMCs) from MS patients spontaneously produce high levels of TNFα , TNFβ , IFNγ, and oncostatin M (oncM), a proinflammatory cytokine acting on cells of neural, vascular, hematopoietic, and lymphoid origin. Spontaneous production of these cytokines was significantly higher (p<0.01) in PBMC short-term culture supernatants from MS patients than in blood donors (HC). On average, lectin-induced production of these cytokines by PBMC was higher in MS patients than in HC, significantly so only for TNFα (p=0.013). Determination of TNFα, TNFβ, IFNγ, and oncM in corresponding sera showed that, on average, oncM levels were higher in MS patients than in HC, though the results were not statistically significant, whereas levels of TNFα, TNFβ, and IFNγ were below the assay threshold in most patients. The finding that MS PBMCs are primed in vivo to produce and release high levels of proinflammatory cytokines suggests the presence of a basal activation of the immune system which, in turn, may play a role in the complex circuitry of molecular and cellular interactions responsible for neurologic damage in MS. [ABSTRACT FROM AUTHOR]

We have previously reported that antipeptide antibodies directed against the second extracellular loop of the cardiac h5-HT 4 receptor could block the activation of the L-type Ca channel in human atrial cardiomyocytes. In this paper we investigate the immunological and physiological activity of these antibodies, in a cell system expressing a larger amount of receptors than the atrial cells. The recombinant receptor was expressed at the surface of COS-7 cells under an active form (serotonin, EC 50 = 1.81 × 10 -7 M), at a high level (375 ± 25 fmol receptor/mg total protein) and was able to bind a specific ligand (GR113808) with a high affinity (Kd = 0.28 ± 0.05 nM). In this system, the same anti-peptide antibodies used for the cardiac cells induced an "agonist-like" effect on the recombinant h5-HT 4 receptor. These results are in line with those shown for others G-protein coupled receptors, as adrenoreceptors. In addition, this work showed that the effect of the antibodies is not only dependent on the epitopic region recognised but also on the molecular density and/or the cellular environment of the target receptors. Finally, our results support the hypothesis that the h5-HT 4 receptor could be a new target for autoantibodies in patients with atrial arrhythmia. [ABSTRACT FROM AUTHOR]

Objective: The northern-Italian region of Piedmont has a total population of 4.3 million. The regional territory is currently divided into 22 local health services, each serving a population not exceeding 300 000 and incorporating a Department of Mental Health. The organization of psychiatric assistance can be described as relatively stable and conforms with the standards imposed by national regulations. Drawing upon previous research experience, and with the support of the relevant regional authorities, our survey implements the methodology developed by the EPCAT team and will eventually embrace about 80% of all local health service areas in this region. Methods: The survey was preceded by a number of preliminary steps including the training of four researchers, inter-rater reliability testing (particularly for the International Classification of Mental Health Care), and the adjustment of the methodology developed by the EPCAT team to facilitate understanding and use of the instrumentation by the psychiatric professionals involved in the data collection. Results: This paper summarizes data on the structure and components of the mental health service in three areas of Piedmont, with special reference to structures funded by the National Health System. Conclusion: Overlooking the imbalances affecting both the resources available for different populations, and the operational profiles of psychiatric agencies of the same type, our data allow an assessment of the descriptive and comparative capabilities of the proposed methodology. [ABSTRACT FROM AUTHOR]

In vitro hemopoiesis and hemopoietic cytokines production were evaluated in 9 centenarians (median age 100.5 years, age range: 100-104 years), 10 old people (median age: 71 years, age range: 66-73 years), and 10 young people (median age: 35 years, age range: 30-45 years), all carefully selected for their healthy status. The main findings were the following: (i) a trend towards a decreased absolute number of CD34+ progenitor cells in the peripheral blood of old people and centenarians, in comparison to young subjects; (ii) a well-preserved capability of CD34+ cells from old people and centenarians to respond to hemopoietic cytokines, and to form erythroid (BFU-E), granulocyte-macrophagic (CFU-GM), and mixed colonies (CFU-GEMM) in a way (number, size, and morphology) indistinguishable from that of young subjects; (iii) an age-related decreased in vitro production of granulocyte-macrophagic colony-stimulating factor (GM-CSF) and a decreased production of interleukin-3 (IL-3) in centenarians by phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC); (iv) a linear increase of the serum level of stem cell factor (SCF), measured in the above-mentioned subjects and in 65 additional subjects, including 4 centenarians. These data suggest that basal hematopoietic potential is well preserved in healthy centenarians, and that the hemopoietic cytokine network undergoes a complex remodeling with age. [ABSTRACT FROM AUTHOR]

We measured the in vitro production of interferon-gamma (IFN-γ) in five cases of hyper-IgE syndrome (HIgE), induced by mitogens, calcium ionophores and phorbol ester. The biosynthesis of IFN-γ was severely reduced or undetectable in HIgE, while it was near normal in most atopic patients. The in vitro spontaneous production of IgE was increased overall in HIgE patients, although no correlation was found with serum IgE levels. Recombinant interleukin-4 (IL-4) induced a further increase in IgE synthesis, and its effect was totally antagonized by recombinant IFN-γ; the same pattern of response was also observed in atopic subjects with high production of IgE. IFN-α synergized with IL-4 on IgE synthesis, whereas recombinant IL-6 gave opposite changes in individual cases tested. We propose that IFN-γ deficiency may be responsible for some of the features of HIgE patients, including IgE levels and infections. [ABSTRACT FROM AUTHOR]

The study of 87 adults of different ages, including 15 centenarians, selected for their healthy status, showed that profound changes of humoral immunity occur throughout life. In particular, a Statistically significant age-related increase of the serum level of immunoglobulin classes (IgG and IgA but not IgM) and IgG subclasses (IgGl, 2 and 3, but not IgG4) was detected. A parallel age related decrease of circulating B cells was also observed. The hypothesis of a complex derangement of B cell function and/or compartmentalization with age is put forward, together with the proposal that healthy centenarians (as representative of successful ageing) may be helpful in identifying the physiological age-related modifications of the immune system. [ABSTRACT FROM AUTHOR]

The authors studied CD40 ligand (CD40L) expression and interleukin-10 (IL-10) production in 16 patients with common variable immunodeficiency (CVI). Mean CD40L expression, determined by using cytofluorimetry, and measured as the mean fluorescence intensity following stimulation of peripheral blood mononuclear cells (PBMC) with phorbol myristate acetate (PMA) and calcium ionophore in 12 patients, was comparable to that of controls. However, three CVI patients showed fluorescence intensity in stimulated cells below 2 standard deviations of normal donors' mean and two other patients had only a slight increase of stimulated versus unstimulated cells (<10 channels). IL-10 production after stimulation of PBMC with both anti-CD3 or anti-CD3 plus PMA gave similar results in CVI patients and normal controls. In vitro stimulation of PBMC with anti-CD40 and various combinations of cytokines (IL-2, IL-4 and IL-10) induced IgG production above 100 ng/ml in one CVI patient out of 13 tested. The data suggest that alterations of IL-10 production are unlikely to play a major role in the pathogenesis of impaired IgG production in most CVI patients. CD40L appears to be normally expressed in two thirds of CVI patients, but it may be functionally defective. [ABSTRACT FROM AUTHOR]

Linked Article:Frew. Br J Dermatol 2019; 180:431–432. [ABSTRACT FROM AUTHOR]

The article presents a case study of two brothers with severe intellectual disability (ID) and congenital microcephaly. It notes that the brothers were unable to take care of their selves and both demonstrated hyperactivity with aggressive behavior. Magnetic resonance imaging (MRI) of the brain has depicted thin brain stem, normal corpus callosum and global reduction in brain size of the older brother. It indicates that several genetic tests were performed after the genetic counseling.

Discusses the case of a female patient with penicillin allergy. Signs and symptoms; Patient medical history; Immunoglobulin E-mediated hypersensitivity to penicillin-G.

Presents the case of an aged female who had itchy dermatitis after an injection with lidocaine hydrochloride. History of illness; Tests administered to the patient.

Reports on a case of chronic urticaria associated with celiac disease with a marked improvement of symptoms after adoption of a gluten-free diet. Cause of gluten-sensitive enteropathy; Tolerance of patients with celiac disease to small amounts of oats.

Interleukin-5 is a T-celi-, mast-cell-, and eosinophil-derived cyt okine with selective effect on cells of the eosinophil lineage. Its role is debatable in experimental models of bronchial hyperresponsiveness, where it is linked to induction of 1gB responses in mice, as well as in human asthma. CD8 + lymphocytes exerting Th2-like activity on B-cell production of IgE have been reported. The unexpected and still unreported finding in this patient was the increased presence of a subpopulation of double-negative.

Background Bullous pemphigoid (BP) is an autoimmune skin disease in which patient autoantibodies react with BP180 and BP230 proteins. In addition to IgG, IgE has been shown to play a role in the disease. Objectives To evaluate the feasibility of detecting IgE and IgG against the immunodominant BP180 NC16A domain (BP180) using a microarray system. Methods BP180 was immobilized on an experimental version of the ISAC® microarray (Exp96). The BP study group and the controls were all tested on the commercial ISAC 103 version and on the Exp96. IgG and IgE were measured in a single run. BP180 IgG and IgE results were compared with those using an enzyme-linked immunosorbent assay (ELISA). Results All results obtained using the IgG ELISA on the 31 patients with BP were replicated with the ISAC IgG. Five of eight BP sera tested by ELISA showed similar results with ISAC IgE. Twenty-nine (94%) and 19 (61%) of the 31 patients with BP were IgG and IgE positive to BP180, respectively, whereas four (3%) and six (4%) of 138 normal donors were IgG and IgE positive, respectively. Interestingly, the levels of IgG against BP180 detected using the ISAC system were related to the disease severity. Patients with BP showed a peculiar profile of IgE recognition toward some groups of allergens, which was absent in a group of allergic individuals. A significant, higher prevalence of hen's egg recognition was observed in patients with BP who had specific IgE to BP180. Conclusions The present preliminary study indicates that the ISAC microarray system is suitable for detecting IgG and IgE autoantibodies in patients with BP. Notably, this system allows the assessment of IgE and IgG autoantibodies at the same time, could be employed for the detection of autoantibodies to other autoantigens, and allows profiling for specific IgE to allergens. [ABSTRACT FROM AUTHOR]

To cite this article: Pfiffner P, Stadler BM, Rasi C, Scala E, Mari A. Cross-reactions vs co-sensitization evaluated by in silico motifs and in vitro IgE microarray testing. Allergy 2012; 67: 210-216. Abstract Background and objective: Using an in silico allergen clustering method, we have recently shown that allergen extracts are highly cross-reactive. Here we used serological data from a multi-array IgE test based on recombinant or highly purified natural allergens to evaluate whether co-reactions are true cross-reactions or co-sensitizations by allergens with the same motifs. Methods: The serum database consisted of 3142 samples, each tested against 103 highly purified natural or recombinant allergens. Cross-reactivity was predicted by an iterative motif-finding algorithm through sequence motifs identified in 2708 known allergens. Results: Allergen proteins containing the same motifs cross-reacted as predicted. However, proteins with identical motifs revealed a hierarchy in the degree of cross-reaction: The more frequent an allergen was positive in the allergic population, the less frequently it was cross-reacting and vice versa. Co-sensitization was analyzed by splitting the dataset into patient groups that were most likely sensitized through geographical occurrence of allergens. Interestingly, most co-reactions are cross-reactions but not co-sensitizations. Conclusions: The observed hierarchy of cross-reactivity may play an important role for the future management of allergic diseases. [ABSTRACT FROM AUTHOR]

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