Impact of individual molecular components in determining primary sensitization to latex.

Allergy to natural rubber latex emerged as one of the main allergies at the beginning among some professional groups and the general population. Sensitization and development of latex allergy have been attributed to exposure to products containing residual latex proteins. The prevailing cross-reactivity of latex proteins with other food allergens is of great concern. Numerous purified allergens are currently available, which greatly help in patient management, thus determining their specific profile. We conducted a multicenter study to investigate changes, from the ROC analysis, in the characteristics of patients with latex allergy by measuring its major protein components. Sensitization to latex proteins is crucial because it highlights the cross reactivity to inhalants (pollen) and food (fruit). It is very essential in an accurate and specific clinical setting.

ICH3, a selective alpha7 nicotinic acetylcholine receptor agonist, modulates adipocyte inflammation associated with obesity.

PURPOSE: The alpha7 nicotinic acetylcholine receptor (α7nAChR), involved in the modulation of inflammation and insulin sensitivity, is downregulated in white adipose tissue (WAT) of obese patients. This study aims to test the ability of a selective synthetic α7nAChR agonist, the spirocyclic Δ2-isoxazoline derivative (R)-(-)-ICH3 (ICH3), to counteract acute inflammation and obesity-associated modifications in WAT. METHODS: We employed the LPS-septic shock murine model, human primary adipocytes and diet-induced obese (DIO) mice. Inflammatory factor expression was assessed by ELISA and quantitative real-time PCR. Flow cytometry was employed to define WAT inflammatory infiltrate. Insulin signaling was monitored by quantification of AKT phosphorylation. RESULTS: In the septic shock model, ICH3 revealed antipyretic action and reduced the surge of circulating cytokines. In vitro, ICH3 stimulation (10 µM) preserved viability of human adipocytes, decreased IL-6 mRNA (P < 0.05) and blunted LPS-induced peak of TNFα (P < 0.05) and IL-6 (P < 0.01). Chronic administration of ICH3 to DIO mice was associated with lower numbers of CD8+ T cells (P < 0.05) and to changed WAT expression of inflammatory factors (Hp, P < 0.05; CD301/MGL1, P < 0.01; Arg-1, P < 0.05). As compared to untreated, ICH3 DIO mice exhibited improved insulin signaling in the skeletal muscle (P < 0.01) mirrored by an improved response to glucose load (ipGTT: P < 0.05 at 120 min). CONCLUSIONS: We proved that ICH3 is an anti-inflammatory drug, able to reduce inflammatory cytokines in human adipocytes and to blunt the effects of obesity on WAT inflammatory profile, on glucose tolerance and on tissue insulin sensitivity.

Response to: Bias in the use of a SSClow/CCR3pos gate to capture basophils in chronic urticaria?

A response to Chirumbolo S, Bjørklund G, and Vella A, Bias in the use of a SSClow/CCR3pos gate to capture basophils in chronic urticaria? Immunobiology, 224(3), 2019, 353-354.

Cord and blood levels of newborn IgE: Correlation, role and influence of maternal IgE.

The role of cord blood immunoglobulin E (IgE) levels in predicting the development of atopy has been widely investigated. The aim of the study was to evaluate the correlation between serum and cord blood total IgE in newborns and the possible influence of the atopic status of the mother on them. It was also investigated the possible role of gestational age on neonatal total IgE levels. We considered 763 deliveries, 724≥37 weeks of gestation and 39<37 weeks of gestation. 14% of mothers (13.7% at term, 15.4% preterm) showed high total IgE levels. The results showed a significant correlation between serum and cord IgE levels both in preterm and term newborns. The data revealed also that mother's total IgE levels affect both neonatal serum and cord total IgE levels. For the latters we also found child gender as an additional independent predictor. On the contrary total IgE levels are not affected by gestational age. Clinical limitations of total IgE is known but their determination can be useful to define atopy and to suggest follow-up of the children.

Profiles of Birch Sensitization (Bet v 1, Bet v 2, and Bet v 4) and Oral Allergy Syndrome Across Italy.

BACKGROUND: Birch allergy (BA) is a common pollinosis caused by the allergens Bet v 1, Bet v 2, and Bet v 4. Oral allergy syndrome (OAS) is frequently associated with BA. A gradient of sensitization to birch allergen across Europe has been reported. Therefore, this study aimed to investigate the birch sensitization profile, including OAS, across Italy. METHODS: We performed a retrospective study of 854 patients (391 males, mean age 35.9 years, range 18-93 years): 196 patients were recruited in Genoa, 188 in northern Italy, 359 in central Italy, and 111 in southern Italy. Serum IgE to Bet v 1, Bet v 2, and Bet v 4 was assessed, and OAS was analyzed. RESULTS: With respect to the geographical path Genoa-North-Center-South, the frequency of sensitization to Bet v 1 decreased significantly (P<.0001) from Genoa (95.41%) to southern Italy (58.56%). The frequency of sensitization to Bet v 2 increased significantly (P<.0001) from Genoa (6.12%) to southern Italy (52.25%). The frequency of Bet v 4 also increased significantly (P=.0002) from Genoa (6.12%) to southern Italy (14.41%). The distribution of patients with OAS differed significantly across the areas (P<.0001), the most marked difference ranging between 33.5% in Genoa and 76.9% in northern Italy. The frequency of birch allergens correlated with OAS in central Italy only. CONCLUSIONS: The present study demonstrated a significant difference between sensitization to birch and its clinical expression across Italy.

Simultaneous detection of circulating immunological parameters and tumor biomarkers in early stage breast cancer patients during adjuvant chemotherapy.

BACKGROUND: Chemotherapy-induced immune suppression has mainly been studied in patients with advanced cancer, but the influence of chemotherapy on the immune system in early stage cancer patients has so far not been studied systematically. The aim of the present study was to monitor the immune system during anthracycline- and taxane-based adjuvant chemotherapy in early stage breast cancer patients, to assess the impact of circulating tumor cells on selected immune parameters and to reveal putative angiogenic effects of circulating endothelial cells. METHODS: Peripheral blood samples from 20 early stage breast cancer patients were analyzed using a flow cytometric multi-color of antibodies to enumerate lymphocyte and dendritic cell subsets, as well as endothelial and tumor cells. An enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of various serological factors. RESULTS: During chemotherapy, all immunological parameters and angiogenesis surrogate biomarkers showed significant decreases. The numbers of circulating tumor cells showed significant inverse correlations with the numbers of T helper cells, a lymphocyte subset directly related to effective anti-tumor responses. Reduced T helper cell numbers may contribute to systemic immunosuppression and, as such, the activation of dormant tumor cells. CONCLUSIONS: From our results we conclude that adjuvant chemotherapy suppresses immune function in early stage breast cancer patients. In addition, we conclude that the presence of circulating tumor cells, defined as pan-cytokeratin(+), CD326(+), CD45(-) cells, may serve as an important indicator of a patient's immune status. Further investigations are needed to firmly define circulating tumor cells as a predictor for the success of breast cancer adjuvant chemotherapy.

TRYPTASE AND HISTAMINE MAY SUPPORT ORAL FOOD CHALLENGE IN THE DIAGNOSIS OF ALLERGY.

Allergen-specific immunoglobulin E (IgE) reactions lead to acute degranulation of mast cells and basophils and release of stored mediators, particularly tryptase and histamine, which can be measured in vitro after reactions. The aim of this study was to investigate the utility of serum tryptase and plasma histamine during oral food challenge (OFC) in 103 children with suspected food allergy, in order to support the diagnosis of a IgE-mediated reaction. Blood samples for serum tryptase and plasma histamine were collected before the OFC and after the onset of allergic symptoms or after 60 minutes from test completion. Serum tryptase and plasma histamine were measured by a fluoroenzyme immunoassay (ImmunoCAP; ThermoFisher, Uppsala, Sweden) according to the manufacturer’s instructions. A correlation between serum tryptase and plasma histamine distributions was observed after OFC (p=0.0035). A correlation was also observed for both serum tryptase and plasma histamine before and after OFC (p less than0.0001). Subjects with positive response to OFC had significantly higher values (p = 0.0375) of serum tryptase compared to subjects with negative response. The plasma histamine distribution showed a significant difference between measurements before and after OFC, both in the complete population (p less than 0.0001), and considering the response (negative OFC: p less than 0.0001; positive OFC: p=0.0181). The diagnostic work-up of IgE- mediated food allergy may include determination of serum tryptase and plasma histamine, in order to support the results of OFC. These markers are strongly related to the same IgE-mediated mechanism and, as they can be both easily measured, can confirm the allergic nature of a reaction in the real-life setting of food allergy.

WALNUT SENSITIZATION IN PEDIATRIC AGE: A PRELIMINARY STUDY.

Walnut consumption has recently become a healthy dietary habit worldwide, due to its positive benefits in reducing cholesterol levels and oxidative stress; this has resulted in an increase in individual consumption, global production and risk of developing sensitization and allergy. In general, clinical manifestations of walnut allergy are frequently severe and systemic potentially life-threatening, leading to anaphylaxis both in the pediatric and adult populations. In light of these findings, we performed a preliminary study considering the walnut native allergen and the recombinant Jug r1 in order to evaluate their role in atopic diseases.

SERUM IMMUNOGLOBULIN IgE IN A PEDIATRIC POPULATION: A RETROSPECTIVE ANALYSIS.

UNLABELLED: Allergic sensitization is mediated by immunoglobulin E (IgE) and an increase of their total value is frequently used to complete a correct diagnosis of atopy. Serum IgE may be considered a typical biomarker for the allergic phenotype. The aim of this study was to evaluate total serum IgE, according to sensitizations and to find a cut off to discriminate between atopic and non-atopic subjects. Seven hundred and ninety-five patients were enrolled in this study. Serum levels of total IgE were measured by a fluorescence immunoassay (ImmunoCAP; ThermoFisher, Uppsala, Sweden) while specific IgE levels were measured by immunofluorometric assay (ImmunoCAP; ThermoFisher, Uppsala, Sweden). Both tests were expressed in kU/L, according to manufacturer’s instructions. RESULTS: A difference for total IgE, according to the gender, has been found (p = 0,0012 ) with higher values for males than for females. A correlation has been found between total IgE and specific IgE, even distinguishing the population in sensitized and non-sensitized. A statistically significant difference has been found according to the presence or the absence of sensitization (p less than 0.0001) and also considering mono-sensitized and polysensitized patients (p less than 0.0001). ROC analysis has been performed to define a cut off for total serum IgE, according to sensitization and to the type of sensitization (mono-sensitization or poly-sensitization). Finally multiple regression models have been performed to describe total IgE response (positive or negative) and to predict total IgE values. Since clinical limitations are well known, total IgE provide a useful aid to define atopy, allowing the clinician to carry out further investigations in patients with total IgE values beyond normal limits.

-295 T-to-C promoter region IL-16 gene polymorphism is associated with Whipple's disease.

Whipple's disease (WD) is a rare systemic condition caused, in genetically predisposed subjects, by Tropheryma whipplei, a common bacterium widespread in the environment. The relevance of genetic predisposition in WD is shown by the association with HLA alleles DRB1*13 and DQB1*06 and by the demonstration that, in patients with WD, the cytokine genetic profile is skewed toward a Th2 and Treg response. Since IL-16 is involved in hampering the development of a protective macrophagic response against Tropheryma whipplei, we investigated whether the genetic background of IL-16 is different between patients with WD and controls. The -295 T-to-C polymorphism of the promoter region of the IL-16 gene was studied in 90 patients with WD and 152 healthy controls. Levels of serum IL-16 protein were also tested. The frequency of the wild type T allele was significantly higher in patients with WD compared to the controls (155/180 vs. 235/304; p = 0.02 for the Chi(2) test), odds ratio 1.82 [95 % confidence interval (CI) 1.07-3.10]. The TT genotype was found in 65/90 patients with WD and 88/152 controls (p = 0.026). No relationship was found between serum levels of IL-16 and genotypes. Although the functional consequences of this genetic background on levels of IL-16 and on the course of the disease are still unknown, we found, for the first time, that the wild type T allele and the TT genotype of the -295 polymorphism are associated with WD.

Pidotimod may prevent recurrent respiratory infections in children.

AIM: Recurrent respiratory infections (RRI) constitute a social problem for both the pharmaco-economic impact and the burden for the family. Pidotimod is a synthetic immunostimulant. The aim of this study was to evaluate the effects of pidotimod on RRI prevention in children. METHODS: Globally, 100 children (49 males, mean age 4.7 ± 1.2 years) with RRI were enrolled in the study. At baseline, children were randomly assigned to the treatment with pidotimod 400 mg/die or not for two months. Children were visited at baseline, after 30 (T1) and 60 (T2) days, and at follow-up (120 days; T3). Number of children with upper and lower airways symptoms, medications use, school attendance, and paediatric visits for RRI were evaluated. RESULTS: Pidotimod treatment was able of significantly reducing the number of children with upper and lower airways symptoms, and medications use, increasing school attendance, and reducing pediatric visits for RRI. CONCLUSION: This study provided the evidence that pidotimod may be able of preventing RRI in children.

Agonists with supraphysiological efficacy at the muscarinic M2 ACh receptor.

BACKGROUND AND PURPOSE: Artificial agonists may have higher efficacy for receptor activation than the physiological agonist. Until now, such 'superagonism' has rarely been reported for GPCRs. Iperoxo is an extremely potent muscarinic receptor agonist. We hypothesized that iperoxo is a 'superagonist'. EXPERIMENTAL APPROACH: Signalling of iperoxo and newly synthesized structural analogues was compared with that of ACh at label-free M2 muscarinic receptors applying whole cell dynamic mass redistribution, measurement of G-protein activation, evaluation of cell surface agonist binding and computation of operational efficacies. KEY RESULTS: In CHO-hM2 cells, iperoxo significantly exceeds ACh in Gi /Gs signalling competence. In the orthosteric loss-of-function mutant M2 -Y104(3.33) A, the maximum effect of iperoxo is hardly compromised in contrast to ACh. 'Superagonism' is preserved in the physiological cellular context of MRC-5 human lung fibroblasts. Structure-signalling relationships including iperoxo derivatives with either modified positively charged head group or altered tail suggest that 'superagonism' of iperoxo is mechanistically based on parallel activation of the receptor protein via two orthosteric interaction points. CONCLUSION AND IMPLICATIONS: Supraphysiological agonist efficacy at muscarinic M2 ACh receptors is demonstrated for the first time. In addition, a possible underlying molecular mechanism of GPCR 'superagonism' is provided. We suggest that iperoxo-like orthosteric GPCR activation is a new avenue towards a novel class of receptor activators.

Adenoid hypetrophy: definition of some risk factors.

Adenoids removed for airway obstruction and-or recurrent infections have been studied to identify a possible mechanism to explain chronicity. In this regard, macrophages may play a relevant pathogenic role as well as neutrophils during bacterial infections and eosinophils in allergic inflammation. Therefore, this study aimed at investigating some mediators as surrogate markers of inflammation in children who had to undergo to adenoidectomy. Globally, 67 children (25 females, 42 males, mean age 4.9 years), affected by persistent obstruction caused by adenoid hypertrophy were consecutively enrolled into the study. Blood samples were collected from patients and controls to determine serum CD163, Myeloperoxidase (MPO) and ECP. There were significant differences between patients and controls for serum CD163 (p less than 0.0001); MPO (p less than 0.0001); serum ECP (p less than 0.0001). This study demonstrated some risk factors for severe AH: apnoea, recurrent respiratory infections, and high serum CD163 levels.

Pentraxin 3 serum levels in children with atopic dermatitis.

PTX3 behaves as an acute-phase response protein as its blood levels rapidly and dramatically increase during endotoxic shock, sepsis, and other inflammatory and infectious conditions. Therefore, this study was designed to investigate a possible role of PTX3 in children with Atopic Dermatitis (AD). One-hundred-and-thirty-six patients (37 females, 99 males, mean age 10.4 years) were enrolled in the study. One hundred patients (74%) had only respiratory symptoms (allergic rhinitis and/or bronchial asthma); thirty-six patients (26%) showed dermatitis associated with respiratory allergy (allergic rhinitis and/or bronchial asthma). PTX3 levels were higher in children with AD and there was a significant correlation between serum PTX3 levels and SCORAD index (p-value=0.0001, rho=0.658). Therefore, this study may show that PTX3 might be a reliable marker for the severity of AD in children with respiratory allergy.

Pentraxin 3 in children suffering from allergic rhinitis.

Allergic disorders are typically characterized by an inflammatory response to allergen exposure. PTX3 behaves as an acute-phase response protein as there is a relationship between PTX3 plasma levels and disease severity. Therefore, this study was designed to investigate a possible role of PTX3 in children with allergic rhinitis. One hundred patients (28 females, 72 males, median age 11 years) were enrolled in the study. All patients were monosensitized: 43 (43%) to seasonal allergens (Graminaceae), 57 (57%) to perennial allergens (house dust mites, cat and dog epithelium, alternaria tenuis). Patients' blood samples for assessing serum PTX3 levels were performed during the spring. Children with rhinitis had higher PTX3 levels and there was a significant relationship between symptom severity and serum levels. Therefore, this study shows that PTX3 serum levels could be a reliable marker for symptom severity in children with allergic rhinitis.

Patients with oral allergic syndrome to apple have intense proliferative response to BET V 1.

Patients with pollen allergy may frequently present an additional food-related allergy (Oral Allergic Syndrome, OAS), as consequence of cross-reactivity between pollen allergens (mainly birch, hazelnut, alder, mugwort) and vegetable allergens. The aim of this study was to evaluate the effect on Bet v 1-induced T cell proliferation exerted by the presence of OAS in birch patients. Fourteen allergic patients were evaluated (6 males, mean age 35.8 years). All of them were monosensitized to birch and suffered from allergic rhinitis: 4 had also OAS to apple. Proliferation of peripheral mononuclear cells was evaluated using Bet v 1 and non-specific stimuli. OAS had higher proliferation than non-OAS patients. In addition, there were significant relationships between immunological and clinical parameters in OAS patients. This study evidences that OAS characterizes a more severe form of birch allergy: as OAS patients had higher SI, circulating eosinophils, and IgE levels. Thus, this study confirms the previous report and underlines the relevance of measuring recombinant birch allergen as higher values may suggest a reliable prediction of OAS.