G. A. Rossi

High serum levels of tumour necrosis factor‐α and interleukin‐8 in severe asthma: markers of systemic inflammation?

Background Severe asthma is characterized by elevated levels of pro‐inflammatory cytokines and neutrophilic inflammation in the airways. Blood cytokines, markers of ‘systemic’ inflammation, may be a feature of amplified inflammation in severe asthma.

Cysteinyl leukotrienes induce human eosinophil locomotion and adhesion molecule expression via a CysLT1 receptor-mediated mechanism

Background The mechanisms involved in eosinophil recruitment by cysteinyl‐leukotrienes (CysLTs) remain to be defined.

Fluticasone and salmeterol downregulate in vitro, fibroblast proliferation and ICAM-1 or H-CAM expression

Beta2-adrenoreceptor agonists have pharmacological properties that may suggest an inhibitory effect on various aspects of the inflammatory and repair processes that characterize asthma. Since fibroblasts express beta2-adrenoreceptors, the effects of different concentrations (0.1-100 nM) of fluticasone propionate (FP), salmeterol (S) and their combination (FP+S) on lung fibroblast proliferation and adhesion molecule expression were evaluated. Stimulation of human foetal lung fibroblasts with a fibrogenic cytokine, basic fibroblast growth factor (bFGF), resulted in a [methyl-3H] thymidine ([3H]TdR) uptake, four-fold higher than that of control cultures (p=0.0001) and was significantly inhibited by S, at all the concentrations tested (0.1-100 nM; p<0.05). No changes in bFGF-induced cell proliferation were observed in the presence of FP (0.1-100 nM; p>0.05, all comparisons). In addition, the association FP+S did not improve the inhibitory activity of S alone (p>0.05, each comparison). An upregulation of intercellular adhesion molecule-1 (ICAM-1) expression was induced by tumour necrosis factor-alpha (TNF-alpha) (p=0.0004), but not by interleukin-4 (IL-4) (p>0.05), while none of the two cytokines were able to increase hyaluronic-cellular adhesion molecule (H-CAM) expression by lung fibroblasts (p>0.05). A significant downregulation of ICAM-1 or H-CAM expression was demonstrated in the presence of FP or S, at all concentrations tested (0.1-100 nM; p<0.01, each comparison). Interestingly, S (10 nM and 100 nM) was able to enhance the inhibitory activity of FP on ICAM-1 expression (p<0.01), but not on H-CAM expression (p>0.1). These results show that in human foetal lung fibroblasts, fluticasone propionate and salmeterol are effective in modulating in vitro, different lung fibroblast biological functions that are likely to be involved in airway remodelling.

Regular vs prn nebulized treatment in wheeze preschool children

Background:  International guidelines recommend regular treatment with inhaled glucocorticoids for children with frequent wheezing; however, prn inhaled bronchodilator alone or in combination with glucocorticoid is also often used in practice. We aimed to evaluate whether regular nebulized glucocorticoid plus a prn bronchodilator or a prn nebulized bronchodilator/glucocorticoid combination is more effective than prn bronchodilator alone in preschool children with frequent wheeze.

Interferon-gamma and IL-10 may protect from allergic polysensitization in children: preliminary evidence.

To cite this article: Prigione I, Morandi F, Tosca MA, Silvestri M, Pistoia V, Ciprandi G, Rossi GA. Interferon‐gamma and IL‐10 may protect from allergic polysensitization in children: preliminary evidence. Allergy 2010; 65: 740–742.

Mechanisms of bradykinin-induced contraction in human fetal lung fibroblasts

Bradykinin (BK) induces fibroblast contraction but the structural changes and intracellular mechanisms involved have not been completely explored. We stimulated HFL-1 fibroblasts with BK to assess: 1) fibroblast contractility; 2) the role of α-smooth muscle actin (SMA) in contraction by small interfering RNA (siRNA); 3) α-SMA protein expression; 4) α-SMA and F-actin structure; 5) intracellular calcium concentration ([Ca2+]i); and 6) phosphorylated myosin light-chain (pMLC) and MLC kinase (MLCK) expression. BK triggered concentration- and time-dependent fibroblast gel contraction in conjunction with α-SMA over expression, but not in α-SMA-siRNA-treated cells. BK also increased α-SMA+ and F-actin+ cell number and stress fibre polymerisation (detectable at 5–60 min). These BK-induced changes were associated with an increase in [Ca2+]i, which peaked within 15 s, and activation of pMLC, which was detectable at 5–60 min. No MLCK content modification was observed. The different manifestations of the BK-induced fibroblast activation were downregulated at different levels (25–100%) by HOE140, a specific BK B2 receptor (B2R) antagonist and by the Ca2+ chelator, EGTA. Thus, BK-induced fibroblast contraction, associated with differentiation into α-SMA+ myofibroblasts, is mediated through the activation of the B2R and involves the Ca2+/calmodulin pMLC-dependent pathway.

Peanut anaphylaxis: the usefulness of molecular-based allergy diagnostics

The peanut is the fruit of the herbaceous plant Arachis hypogea belonging to the Fabaceae (leguminous) family. The fruit (peanut) arises as a leguminous growing underground (hypogea), containing 2–3 seeds, commonly eaten roasted or worked to obtain oil or butter. The peanut allergy (PA) prevalence is about 1–2% in children, and PA may account also for severe food allergy (1). The severity of reactions may depend on the type of allergen component sensitization. In this regard, at least 11 peanut allergen proteins have been identified and characterized (2– 4). Ara h 1, 2 and 3 are the major peanut allergens belonging to the seed storage proteins family (Ara h 1 is a vicilin, Ara h 2 is a conglutin, Ara h 3 is a glycinin). Ara h 4 is a glycinin; Ara h 5 is a profilin. Ara h 6 and 7 are similar to Ara h 2. Ara h 8 is a PR-10 protein. Ara h 9 is a lipid transfer protein (LTP). Ara h 10 and 11 are oleosins. However, Ara h 2 is the most important peanut allergen component and positivity to Ara h 2 defines a ‘true’ PA (2). The PA diagnostic work-up is based on history, detection of allergen-specific IgE by skin prick test and/or serum assessment, and oral challenge. The raw allergen extract is available since much time. However, molecular-based allergy diagnostics allows to define and to characterize the sensitization profile, so identifying potential dangerous proteins and suggesting a more precise prognosis and improving the management. Very recently, Klemans et al. (5) reported that the diagnostic value of sIgE to Ara h 6 on adults was as good as to Ara h 2 PA. However, reported anaphylaxis was not considered in that study. Therefore, the aim of this study was to compare children with peanut anaphylaxis to children with less severe PA investigating the role of molecular-based allergy diagnostics. We retrospectively considered 14 children with history of peanut anaphylaxis and 15 children with diagnosed PA. Specific IgE to raw peanut allergen was measured by ImmunoCAP method (Thermo Fisher, Milan, Italy). Specific IgE to molecular components of peanut, such as Ara h 1, Ara h 2, Ara h 3, Ara h 6, Ara h 8 and Ara h 9, was assayed by the semiquantitative ISAC method (Thermo Fisher). Allergen-specific IgE levels to raw peanut were significantly higher in the anaphylaxis group (P = 0.0044) (Table 1). Sensitization to either Ara h 2 or Ara h 3 was more frequently detected in the anaphylaxis group (P = 0.0209 and P = 0.0007, respectively). Receiver operating characteristic analysis showed that raw peanut cut-off > 2.1 kUA/l, Ara h

TNF-alpha, IL-4R-alpha and IL-4 polymorphisms in mild to severe asthma from Italian Caucasians.

Asthma is a chronic airway inflammatory disease associated with airway hyperresponsiveness which affects subjects with genetic predisposition. An association has been reported between some polymorphisms in various cytokine genes and asthma. Most of them are single nucleotide polymorphisms (SNPs). These polymorphisms are detected in the protein coding sequence or in the promoter region thus influencing cytokine production. We investigated the involvement of SNP mapping in 5 cytokine genes in mild to severe asthmatics of Italian Caucasians. The frequency of alleles and genotypes, relatively to 10 allelic specificities of the cytokine genes, was defined in 57 asthmatics and in 124 control subjects by a Polymerase Chain Reaction-Sequence Specific Primer method. TNF-α -308A and TNF-α -238A allele frequencies were higher in asthmatics than in controls (p<0.001). Significant differences in the frequency of IL-4 -590T allele and of IL-4Rα + 1902A allele were also detected in asthmatics in comparison with controls (p<0.001 and p=0.005, respectively). Similarly, IL-1α -889C allele was present in 84.1% of asthmatics and in 70.2% of controls (p=0.013). Furthermore, the IL-4Rα + 1902A/A and IL-1α -889C/C homozygous conditions and the TNF-α -308G/A, TNF-α -238G/A, IL-4 -590T/C and IL-10 -1082G/A heterozygous conditions were significantly associated with asthma (p<0.05). ACA haplotype of IL-10 was observed only in asthmatic patients. This study reports, for the first time, the frequency of 10 different single nucleotide polymorphisms in 5 cytokine genes in the Italian Caucasians. Furthermore, we also indicate that in our population some single nucleotide polymorphisms are associated with mild to severe bronchial asthma.

Acute anaphylactic reaction after prick-by-prick testing for pine nut in a child.

e nuts are the seeds of Pinus pinea which are a fundantal ingredient of a variety of sauces, including ‘‘pesto’’. sto originated in Genoa, Italy, and the name comes from Italian word ‘‘pestare’’ (to pound or to bruise) since traditional way of making this sauce was with mortar d pestle. Typically, pesto is prepared with fresh basil, rlic, pine nuts, grated parmesan and sheep milk cheese, tra-virgin olive oil, and coarse salt. prick-by-prick testing, nasal and conjunctival itching was observed with congestion. Itching became rapidly systemic, associated with irritability, asthenia, cough and dyspnoea. The clinical examination demonstrated wheezing, SaO2 was 96% in room air and the blood pressure 90/65 mmHg (before testing 110/70). Symptoms rapidly remitted after adrenaline aerosol (10 mg) and parenteral antihistaminic (chlorfenamine 5 mg) and steroidal (betamethasone 1.2 mg) treatment. Anyway, the test was positive: wheal diameter was 4 mm × 4 mm. Further, we obtained the results of the level of serum IgE specific for pine nut 32 kU/L (ImmunoCap System, Phadia, Milan, Italy), thus confirming the IgE-dependent reaction. Acute anaphylactic reaction to pine nut following skin 9 tes bu pri tem

The impact of age on serum allergen-specific IgE to inhaled molecular components

BACKGROUND Respiratory allergy is characterised by an IgE-mediated reaction. The immune system functions, including IgE production, progressively decline over time, such as growing up and ageing. Molecular-based allergy diagnostic defines sensitisation profile. This study aimed to evaluate the impact of age on serum allergen-specific IgE to molecular component levels in a large sample of subjects. METHODS Serum IgE to: Phl p1, Bet v1, Ole e1, Cup a1, Par j2, Can f1, Der p2, and Fel d1 were assessed by ISAC method. Sera from 2788 patients, 1230 males (44.1%) and 1558 females (55.9%), median age 23 years (1st and 3rd quartiles: 9.7-49.7 years; age range: 1 month-103 years) were analysed. RESULTS The number of positive tests (i.e. sensitisation) tended to increase between birth and school-age till young adulthood and then decreased (p<0.0001) with the exception of Fel d 1 (p=0.14). A similar age-dependent trend was observed considering the levels of each allergen components: the levels of each allergen component, with the exception of Fel d 1, tended to increase till early adulthood and then to decrease reaching the lowest levels in the elderly. CONCLUSIONS Allergen-specific IgE production to inhaled molecular components trend to reduce with ageing, but with differences between allergens. This phenomenon should be adequately evaluated managing allergic patients.