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To characterize the cellular inflammation at the bronchial and bronchoalveolar levels, we evaluated 43 patients with asthma who were sensitized to house dust mites. On 2 consecutive days patients underwent methacholine challenge and allergen bronchial challenge. In addition, 6, 24, or 72 h after allergen challenge, fiberoptic bronchoscopy with bronchial lavage (BL) and bronchoalveolar lavage (BAL) was performed. Patients belonging to the 6-h, 24-h, or 72-h group were divided further into two subgroups: those with isolated early response to allergen (LAR&sup-; ), and those with dual response to allergen (LAR&sup+; ). The percentage of eosinophils and of epithelial cells in BAL fluid was significantly higher in LAR&sup+; than in LAR&sup-; patients in the 6-h group (p < 0.05, each comparison), but not 24 or 72 h after (p > 0.05, each comparison). Similarly, the proportion of BL eosinophils was also higher in LAR&sup+; than in LAR&sup-; patients, both in the 6-h and in the 24-h group (p < 0.05, each comparison). In addition, increased proportions of BL neutrophils were present in the LAR&sup+; patients belonging to the 24-h group (p < 0.05). Comparing 'proximal' 4BL vs 'distal' 4BAL data, we found a significantly higher proportion of epithelial cells in BL compared with BAL, in both LAR&sup-; and LAR&sup+; subjects, either 6, or 24, or 72 h after challenge (p < 0.01, each comparison) and increased percentages of BL neutrophils and eosinophils in LAR&sup+; patients (p < 0.05, each comparison), but not in LAR&sup-; patients, in the 24-h group. The percentages of BL or BAL macrophages and lymphocytes did not differ significantly among the different patient groups. These data indicate that the development of LAR after allergen inhalation challenge is associated with an early recruitment of eosinophils and with epithelial desquamation in the airways. In addition, after allergen challenge epithelial desquamation is more pronounced in. [ABSTRACT FROM AUTHOR]

Despite its clinical efficacy on intracellular pathogens, the in-vitro intracellular antimicrobial activity of azithromycin, has been shown to be absent or lower than expected from the intracellular concentrations reached. To test the possibility that the high intracellular concentrations of the drug could damage phagocytes, the present study evaluated the effects of azithromycin on (a) the intracellular killing of Staphylococcus aureus by human blood neutrophils (PMNs) and (b) the viability and the respiratory burst of PMNs. Using a fluorochrome assay, we assessed the phagocytosis and intracellular killing of S. aureus by PMNs preloaded with azithromycin, or by PMNs unloaded but with the drug in the culture medium. In addition, possible drug-induced damage to PMNs was evaluated measuring: (a) hydrogen peroxide (H2O2) production and (b) the percentages of PMNs dead at the end of the phagocytosis process. Compared to control PMNs without drug, a time-dependent enhancement in the intracellular killing was observed which was statistically significant after 60 min incubation. The increased intracellular killing was higher in suspensions of unloaded PMNs and azithromycin (P < 0.01) that in suspensions of preloaded PMNs (P < 0.05). This increased intracellular killing was not associated with increased proportions of dead phagocytes, either in preloaded or unloaded PMNs (P < 0.05, each comparison). Similarly no changes in the production of H2O2 by PMNs were observed in the presence of azithromycin. Thus, azithromycin induces a time-dependent increase in the bactericidal activity of human PMNs, without increasing the phagocyte self-killing or modifying H2O2 production. [ABSTRACT FROM AUTHOR]

Blood levels of inhaled corticosteroids are significantly lower than those measured in the lung, but their concentration could still have anti-inflammatory effects. To determine whether budesonide, at concentrations similar to those obtained in blood after drug inhalation (10-9 M), could downregulate the allergen-induced activation of mononuclear cells, we studied 21 atopic patients, sensitized to Dermatophagoides pteronyssinus (Der p). On blood mononuclear cells, isolated from these patients, incubated with Der p allergen extract and with or without budesonide, we evaluated: 1) the proliferative response of T cells; 2) the expression of two surface activation markers, the HLA-DR antigens and the interleukin (IL)-2 receptors; and 3) the release of cytokines known to modulate the allergic processes. Allergen-induced T-cell proliferation was associated with increased HLA-DR antigen and IL-2 receptor expression (P< 0.001), and with increased release of IL-2, interferon-γ (IFN-γ;), IL-1 β, tumor necrosis factor-alpha (TNF-α), and granulocyte/macrophage colony-stimulating factor (GM-CSF). The addition of budesonide at the beginning of the cell cultures induced a dose-dependent inhibition of T-cell proliferation, still significant (P< 0.05) at the lowest concentrations tested (10-9 and 10-10 M). A significant inhibitory effect on T-cell proliferation was also present when budesonide (10-9 M) was added to the cell cultures 3 or 5 days after the beginning of the cell cultures. [ABSTRACT FROM AUTHOR]

CD40/CD40 ligand interaction is an important pathway for B and T cell cooperation and function; functional CD40 molecules have recently been found on nonhematopoietic cells. We detected CD40 in vivo on normal human respiratory epithelial cells and showed that its expression is increased on inflamed airway epithelium. Subsequently, we analyzed its expression and function on primary cultures of human airway epithelial cells. Our data show that CD40 is up-regulated by IFN-beta and IFN-gamma, its ligation increases the surface expression of CD54 and CD106 and it may stimulate the release of IL-6 and IL-8. The use of Janus kinase 3 (JAK3) and NF-kappaB inhibitors suggests that both basal and CD40-induced release of the two cytokines is JAK3-dependent. Using colocalization techniques, we revealed the existence of CD40/JAK3 and CD40/TNFR-associated factor 2 interplay. The extent of these interactions may be partial (2-40% of the cells) or massive (80-90% of the cells) in cultured cells. Stimulation via CD40 causes a significant increase in the number of cells expressing colocalization only in the cultures displaying low frequency of initial colocalization. Thus, airway epithelial cells, activated by CD40, may behave as effector cells of the inflammation process and should be considered priority targets for anti-inflammatory therapy. This work identifies CD40 and the correlated JAK3 signaling molecule as potential molecular targets to block the inflammatory functions of epithelial cells.

Asthma is characterized by an irreversible subepithelial fibrosis with the appearance of myofibroblasts, which can be now considered important early participants in inflammatory responses as well as potential targets for anti-inflammatory drugs. In this study, we show that fluticasone propionate (FP), a powerful inhaled corticosteroid (ICS), displays novel anti-inflammatory effects on human lung fibroblasts during their myofibroblastic differentiation. Indeed, FP inhibits in lung myofibroblasts, at a very early stage of differentiation, the activation of Janus kinase/STAT pathways induced by IL-13 (tyrosine kinase 2, STAT1, STAT3, STAT6, mitogen-activated protein kinase). Contrarily, in mildly or fully differentiated myofibroblastic cultures, FP still displays a potential anti-inflammatory activity even if it only inhibits tyrosine kinase 2 phosphorylation. Moreover, FP inhibits constitutive and TGF-beta-induced expression of alpha-smooth muscle actin, the main marker of myofibroblastic differentiation, both in very early and in mild differentiated myofibroblasts. Finally, FP displays an additional powerful anti-inflammatory effect, decreasing nuclear translocation of NF-kappaB independent of the degree of myofibroblastic differentiation. These data 1) suggest that myofibroblasts are priority targets for ICS, which is able to revert them to a normal phenotype even if they appear to be already engaged in their differentiation, and 2) may help to explain why asthma is improved by an early ICS treatment, whereas advanced asthma is more resistant to these drugs.

We studied whether different bronchial responses to allergen in asthma and rhinitis are associated with different bronchial inflammation and remodeling or airway mechanics. Nine subjects with mild asthma and eight with rhinitis alone underwent methacholine and allergen inhalation challenges. The latter was preceded and followed by bronchoalveolar lavage and bronchial biopsy. The response to methacholine was positive in all asthmatic but in only two rhinitic subjects. The response to allergen was positive in all asthmatic and most, i.e., five, rhinitic subjects. No significant differences between groups were found in airway inflammatory cells or basement membrane thickness either at baseline or after allergen. The ability of deep inhalation to dilate methacholine-constricted airways was greater in rhinitis than in asthma, but it was progressively reduced in rhinitis during allergen challenge. We conclude that 1) rhinitic subjects may develop similar airway inflammation and remodeling as the asthmatic subjects do and 2) the difference in bronchial response to allergen between asthma and rhinitis is associated with different airway mechanics.

beta2-adrenoreceptor agonists have the ability to downregulate in vitro the proliferative response of peripheral blood mononuclear cells (BMCs). This activity could be related to a variety of beta2-adrenoreceptor-mediated functions, including induction of cell apoptosis in activated T-cells. To test this hypothesis, BMCs from atopic subjects, sensitized to house dust mites (Dermatophagoides [Der p]) and/or to Parietaria were incubated with fenoterol (10(-8)-10(-5) M) in the presence of (a) purified allergen extracts (Der p [5 microg/mL] or Parietaria [5 microg/mL]) or (b) antigens (tetanus toxoid [1 microg/mL] or Candida albicans [5 x 10(5) bodies/mL]). The BMC proliferation was assessed by [3H] thymidine incorporation and cell apoptosis was assessed by evaluating DNA fragmentation by a fluorescence technique, using propidium iodide. In cultures stimulated with Der p or with Parietaria, fenoterol induced a dose-dependent inhibition of BMC proliferation, significant also at the lowest concentration tested (10(-8) M) (p < 0.05, each comparison). In contrast, the inhibitory activity of the drug on tetanus-toxoid-stimulated BMCs was significant only at the highest dose tested (10(-5)M) (p < 0.05), whereas no effect was seen when BMCs were stimulated with C. albicans extract (p > 0.05). The different inhibitory efficacy of fenoterol appeared to be related to the degree of activation of beta2-adrenoreceptors on the different BMC populations that responded to the different stimuli. Indeed, in the presence of fenoterol (10(-6) and 10(-5)M), a significant increase in cyclic adenosine monophosphate (cAMP) levels was seen in Der p- or Parietaria-stimulated cells (p < 0.05; each comparison), but not in cell cultures stimulated with tetanus toxoid or with C. albicans extracts (p > 0.05; each comparison). Finally, the percentage of cells with fragmented DNA was lower in cultures stimulated with Der p or Parietaria than in those stimulated with tetanus toxoid or C. albicans, and the presence of fenoterol did not modify cell apoptosis (p > 0.05; each comparison). Thus, the different inhibitory activity of fenoterol on BMCs activated by allergens (Der p or Parietaria) or by antigens (tetanus toxoid or C. albicans) seems to be related to differences in beta2-adrenoreceptor expression and/or function in the different antigen-specific T-cell subsets, but it is not influenced by changes in cell apoptosis.

Objective: To investigate whether the state of activation of circulating T-cells in childhood asthma could be related to serum IgE levels and/or to blood eosinophilia., Methods: Seventeen atopic asthmatic children, sensitized to Dermatophagoides pteronyssinus (Der p), in stable condition at the time of the study and 15 sex-matched and age-matched controls were studied. The expression of activation surface markers (HLA-DR and CD25) on peripheral blood mononuclear cells (PBMCs) was tested by monoclonal antibodies and FACS analysis, while the PBMC proliferative response to Der p antigens was measured by tritiated thymidine (3HTdR) incorporation., Results: As compared to controls, atopic children showed higher eosinophil counts (P < .01), similar lymphocyte counts (P > .1, each comparison) but higher proportion of HLA-DR+ and CD25+ T-lymphocytes (P < .05, each comparison). A significant Der p allergen-induced PBMC proliferation was observed in atopic children (P < .01) but not in controls (P > .1). Both in controls and in atopic children, no correlations were found between lymphocyte counts and eosinophil counts or total or allergen-specific IgE levels (P > .1, each comparison). In contrast, weak correlations were detected between the degree of allergen-induced PBMC proliferation and: a) allergen-specific IgE levels in serum (P < .05) and b) eosinophil counts (P < .05)., Conclusion: These data support the concept that the degree of activation of allergen-specific T-lymphocytes in blood may reflect the intensity of allergic sensitization in childhood asthma.

Patients with chronic obstructive lung disorders often show increased susceptibility to airway infections. As beta 2-adrenoceptor agonists, in addition to reversing the contractile response of bronchial smooth muscles, may inhibit a variety of inflammatory and immuno-effector cell functions, it is possible that these drugs interfere with host defence mechanisms. The present study was designed to test in vitro whether fenoterol, a short-acting beta 2-adrenoceptor agonist, could modify human blood neutrophil recruitment and antimicrobial activity. Pre-exposure to fenoterol significantly reduced neutrophil migration towards the complement component C5a, at concentrations ranging from 10(-7) M to 10(-5) M, or towards lipopolysaccharide, at a concentration of 10(-5) M (P < 0.05, each comparison). In contrast, the drug (10(-8)-10(-5) M) did not significantly modify the increased expression of lymphocyte function-associated antigen (LFA-1, i.e. CD11a/CD18) the macrophage antigen-1 (Mac-1, i.e. CD11b/CD18) induced by N-formylmethionylleucylphenylalanine (fMLP) (P > 0.05, each comparison). Finally, incubation of neutrophils with fenoterol (10(-8)-10(-5) M) did not significantly influence phagocytosis or intracellular killing of bacteria (Staphylococcus aureus) or H2O2 release induced by tetradecanoyl-phorbol-acetate (P > 0.1 for each comparison). These results suggest that short-acting beta 2-adrenoceptor agonists, such as fenoterol, are able partially to reduce neutrophil recruitment in the airways without interfering with the processes involved in phagocytic activity against bacteria.

Background: Alveolar macrophages (AMs) are more efficient antigen-presenting cells in allergic individuals than in nonatopic subjects., Objective: We studied whether this difference may be correlated to increased expression of membrane costimulatory molecules, such as the B7 molecules (CD80 and CD86)., Methods: Eleven subjects with allergic asthma sensitized to Dermatophagoides pteronyssinus and 5 healthy nonatopic volunteers underwent bronchoalveolar lavage, and the costimulatory molecule expression on AMs was evaluated. Peripheral blood T cells, either freshly isolated or as established D pteronyssinus -specific cell lines, were cultured with autologous monocytes or AMs as antigen-presenting cells. In vitro allergen-induced proliferation and cytokine production were evaluated in the presence of B7-blocking reagents., Results: Allergic individuals had a significantly higher proportion of AMs expressing the CD80 molecule than control subjects (28.5% +/- 14.8% vs 1.4% +/- 1.2%; P <.001), whereas no difference was observed in CD86 expression (2.0% +/- 2.3% vs 1.1% +/- 0.6; P >.1). In a large proportion of the asthmatic subjects we studied, AMs were presenting soluble antigens (tetanus toxoid and streptolysin-O) to freshly isolated T cells more efficiently than AMs from nonatopic control subjects. Finally, both T-cell proliferation and cytokine production of D pteronyssinus- specific established T-cell lines were inhibited by a CD80-blocking antibody in a dose-dependent manner., Conclusion: Costimulation by means of CD80 expressed by AMs is probably involved in the amplification of the allergen-specific T-lymphocyte response in the airways of asthmatic subjects.

In allergic asthma, there is convincing evidence that changes in eosinophil and lymphocyte state of activation in blood may reflect disease activity. We evaluated whether simple blood eosinophil or lymphocyte counts in atopic children with asthma could reflect the degree of allergic sensitization. Seventy-six asthmatic children, sensitized to house dust mites (HDM), in stable conditions at the time of the study, and 53 sex- and age-matched controls (CTR) were studied. As compared to CTR, allergic patients showed higher eosinophil numbers and percentages (p < 0.001) but similar lymphocyte numbers and proportions (p > 0.1). Both in CTR and in allergic patients, eosinophil counts did not correlate with lymphocyte counts (p > 0.05; each comparison) but positive correlations were observed between eosinophil numbers and percentages and paper radio immunosorbent test (PRIST) levels or radio-allergo sorbent test (RAST) classes (p < 0.001; each comparison). When allergic asthmatic individuals were subdivided according to their age into two subgroups (Gr), no differences were found in eosinophil and lymphocyte counts and in PRIST levels and RAST values between Gr1 (< or =5 years old [preschool children]) and Gr2 (>5 years old [school children]) (p > 0.05; each comparison). Interestingly, although positive correlations between eosinophil counts and PRIST levels were found in both subgroups (p < 0.05; each comparison), only in Gr2 did eosinophil counts correlate positively with RAST classes (p < 0.001). No correlations between lymphocyte counts and PRIST levels or RAST classes were demonstrated (p > 0.05; each comparison). These data suggest that although blood eosinophilia was similar in preschool and in allergic asthmatic school children sensitized to HDM, only in the oldest children did blood eosinophil counts appear to be related to the degree of HDM-specific sensitization.

Inflammatory airway disorders, such as asthma and chronic bronchitis, are characterized by overexpression of adhesion molecules on airway epithelial and endothelial cells. This phenomenon is associated with increased adherence and activation of polymorphonuclear leukocytes (PMNs). With the knowledge that beta2-adrenoceptor agonists demonstrate some anti-inflammatory activity in vitro, the present study was designed to evaluate whether fenoterol could interfere with adhesion molecule expression on airway epithelium. Human bronchial epithelial cells (HBECs), obtained by protease digestion from surgically resected bronchi, were stimulated with human recombinant interferon-gamma (rh IFN-gamma) in the presence of (a) fenoterol (10(-12)-10(-5) M); (b) dexamethasone (10(-12)-10(-5) M); and (c) fenoterol and dexamethasone. Because desensitization after high-dose exposure to agonists has been described for many membrane-associated receptors, in additional sets of experiments HBECs were preexposed to fenoterol and, as control, to dexamethasone for 8 hr, then washed and stimulated with rh IFN-gamma in the presence of fresh drugs. The cells were harvested after 24-hr culture and stained by specific monoclonal antibodies. The intensity of intercellular adhesion molecule-1 (ICAM-1) expression was then measured by flow cytometry analysis and expressed as mean fluorescence channel (mfc). The significant increase in ICAM-1 expression on HBECs induced by rh IFN-gamma was inhibited, in a dose-dependent manner, by the two drugs, but fenoterol was more efficient than dexamethasone at all of the concentrations tested (p < 0.05, all comparisons). In addition, the inhibitory activity of fenoterol was not enhanced by the simultaneous presence of dexamethasone in rh IFN-gamma-stimulated HBEC cultures (p > 0.05, all comparisons). Finally, preexposure to fenoterol or to dexamethasone did not induce any modification of the inhibitory effect of the two drugs on ICAM-1 expression (p > 0.05, all comparisons). These results suggest that clinical efficacy of fenoterol in patients with obstructive lung disease may include downregulation of adhesion molecule expression on airway epithelial cells.

Objective: The purpose of this study was to investigate the frequency of allergy in 85 families of pediatric patients with asthma and/or rhinitis., Methods: Families enrolled were drawn according to a table of randomization, from those whose children were referred the outpatient clinic of the Pulmonology Department of Gaslini Institute (Genoa, Italy). In patients and in both their mothers and fathers, allergic sensitization to the three most common classes of inhalant allergens (house dust mites, pollens, and animal danders) was evaluated by skin prick test., Results: As compared with their parents, children showed a similar prevalence of positive skin prick test (50.0% and 58.8%, respectively; chi 2 = 1.4; P > .1), but a significantly higher frequency of specific sensitization to house dust mites, pollens, or pets (P < .05, each chi 2). These differences were at least partially related to the higher frequency of polysensitization (i.e., sensitization to more than one class of allergens) in children than in their parents [chi 2 = 4.2, odds ratio (OR) = 2.3, 95% confidence interval (CI95%) = 1.0 to 5.1; P < .05). In addition, both in children (chi 2 = 5.8, OR = 11.3, CI95% = 1.4 to 81.5; P < .05] and in their parents (chi 2 = 7.4, OR = 4.8, CI95% = 1.5 to 16.7; P < .01), allergy to pollens was less frequent in monosensitized than in polysensitized families, while the prevalence of sensitization to house dust mites was similar in monosensitized and in polysensitized families. The analysis of the role of parental sensitization in the development of allergy in the offspring demonstrated that the prevalence of allergic children: (1) seemed to be higher, without reaching statistical significance, in families with sensitized parents than in those with non-sensitized parents (chi 2 = 0.41, P > .1) and (2) was higher in families with one or both polysensitized parents than in those with one or both monosensitized parents (chi 2 = 4.5, OR = 4.0, CI95% = 1.1 to 15.6; P < .05). In addition, within each family, the coincidence of sensitization to house dust mites was more frequent than that to pollens (60.0% and 36.0%, respectively; chi 2 = 7.6, OR = 4.5, CI95% = 1.5 to 14.3; P < .01), and it was not influenced by the number of parents sensitized to same allergen., Conclusion: These data support the concept that, in addition to genetic predisposition, other factors (i.e., environmental exposure) may influence the development of specific sensitization in children with respiratory symptoms. The different sensitization found in this study between children and their parents occurred in a generation span, so as it is more likely to be related to environmental changes than to genetic factors.

Although neutrophil number may be increased in the airways of patients with asthma, its pathogenetic role in this disorder remains unclear. We evaluated BAL of 8 normal control subjects, 30 +/- 2 years of age, and 24 patients with mild asthma: 17 patients with allergic asthma, 24 +/- 1 years of age, and 7 patients with nonallergic asthma, 30 +/- 1 years of age. The BAL of asthmatic patients showed increased numbers of neutrophils (p < 0.01), eosinophils (p < 0.01), and ciliated epithelial cells (p < 0.05) and increased concentrations of myeloperoxidase (MPO) (p < 0.01) compared with control subjects. Positive correlations were observed between the number of BAL neutrophils and eosinophils (Rs = 0.780, p < 0.0001) and between BAL neutrophil numbers and BAL MPO levels (Rs = 0.40, p < 0.05). No correlations were found between the following: (1) BAL eosinophils or neutrophils and BAL epithelial cells (p > 0.05, each comparison); (2) BAL neutrophils or eosinophils and log Pd15 methacholine (MCh) (p > 0.05, each comparison); or (3) BAL epithelial cells or log Pd15 MCh and BAL MPO (p > 0.05, each comparison). Dividing the patient population into two groups, allergic asthmatics and nonallergic asthmatics, similar BAL neutrophil, eosinophil, and epithelial cell numbers and similar MPO levels were found (p > 0.05, each comparison). In addition, the correlations between BAL neutrophils and eosinophils showed similar significance in the two patient subgroups (p > 0.05, each comparison). These results suggest that, both in allergic and nonallergic asthma, airway recruitment and activation of neutrophils occur as does parallel eosinophil migration. However, airway neutrophils do not seem to contribute significantly to epithelial cell injury or to airway hyperresponsiveness in the steady state.

Background: Allergy is one of the most common causes of respiratory symptoms in children and youth., Objective: Evaluate the presence and the type of allergic sensitization in a paediatric population with respiratory symptoms., Methods: We studied 564 consecutive children, 5 months to 17 years of age, with a male to female ratio (M/F) = 1.4, referred to our outpatient clinic in a 12-month period retrospectively. Patients were arbitrarily divided into four groups (grs) according to their age: gr1 = 5 months to 4 years old (181 patients), gr2 = 4 to 7 years (201 patients), gr3 = 7 to 10 years (96 patients), and gr4 = 10 to 17 years (86 patients). Sensitization to house dust mites, pollens, animal dander, and molds was determined by skin prick testing., Results: Sensitization to at least one class of allergen occurred in 304 of the 564 patients (53.9%, M/F ratio = 2.0); the percentage of allergic patients increased with age as follows: 29.8% (54 patients) of the patients in gr1, 55.2% (111 patients) in gr2, 68.8% (66 patients) in gr3 and 84.9% (73 patients) in gr4 (chi(2) = 84.1, P < .01). In the entire allergic population and in gr1 to gr3, the most common positive allergic reaction was to house dust mites (P < .01, chi(2) test each comparison). In contrast, gr4 patients showed a nearly equal percentage of sensitization to pollens and to house dust mites (79.5% and 78.1% respectively) (chi(2) = 0.0, P = >.1). Sensitization to only one class of allergen occurred in 51.3% of the allergic patients and the percentage of these monosensitized patients tended to decrease from gr1 to gr4 (chi(2) = 15.2, P < .1). In the monosensitized group, sensitization to house dust mites was the most frequent in gr1 to gr3 (age <10 years) as in the whole sample. In gr4, the frequency of sensitization to house dust mites was similar to that of sensitization to pollens. On the contrary, within the patient group sensitized to two or more allergens (polysensitized patients), sensitization to house dust mites was as frequent as sensitization to pollens already in gr2 as compared with monosensitized patients., Conclusions: In children with respiratory symptoms, the percentage of allergic individuals was high and increased with the age of the patients. This phenomenon was associated with an age-related enhancement in the ratio of polysensitized to monosensitized patients and with an age-related increase in the frequency of sensitization to seasonal allergens (ie, pollens).

Background: Corticosteroids are thought to be effective in the treatment of allergic reactions including bronchial asthma because they not only have anti-inflammatory effects, but also downregulate the processes of T-cell activation., Objective: To evaluate in vitro the inhibitory activity of budesonide, a widely used inhaled corticosteroid, on allergen-induced mononuclear cell activation., Methods: Thirty-one atopic asthmatic patients, sensitized to Dermatophagoides pteronyssinus (Der p) were studied. Peripheral blood mononuclear cells isolated from these patients were used to determine the ability of budesonide to inhibit (1) the proliferative response of blood T-lymphocytes to Der p allergen extract and to phytohemoagglutinin (PHA) and (2) the release of different cytokines known to modulate the interaction between T-lymphocytes and monocytes in the allergic processes., Results: A significant T-cell proliferation was observed both in the presence of PHA (P < .001) and that of Der p allergen extract (P < .05), and was associated with increased release of interleukin-2 [IL-2 (respectively P < .001 and P < .01)], gamma-interferon [gamma-IFN (respectively P < .001 and P < .01)], granulocyte-macrophage colony-stimulating factor [GM-CSF (respectively P < .01 and P < .001)], interleukin-1 beta [IL-1 beta (respectively P < .05 and P < .01)], and tumor necrosis factor-alpha [TNF-alpha (P < .05 each comparison)]. The addition at the beginning of the cell cultures of different concentrations (from 10(-10) M to 10(-7) M) of budesonide, and as control of dexamethasone, induced a dose-dependent inhibition of T-cell proliferation, in response to PHA and Der p. Budesonide at the lowest concentrations tested (10(-10) M and 10(-9) M) was more effective than dexamethasone. Budesonide was also more active than dexamethasone in inhibiting the release of IL-2, gamma-IFN, IL-1 beta and GM-CSF (in both PHA-stimulated and Der p-stimulated blood mononuclear cell cultures) and TNF-alpha (in Der p-stimulated blood mononuclear cell cultures)., Conclusions: Budesonide is equally or more effective than dexamethasone in inhibiting the allergen-induced T-cell proliferation and in reducing the release of cytokines by allergen-stimulated blood mononuclear cells.

Chronic bronchitis (CB) is characterized by inflammatory changes in the bronchial tissue and by recurrent bronchitis exacerbations. In addition, defective systemic and local immune mechanisms have been demonstrated and biologic response modifiers (BRMs) have been recently introduced for clinical use in patients with CB. We studied 24 patients with CB by bronchoalveolar lavage (BAL), before and after a 4-week treatment protocol with inhaled Immucytal (Pierre-Fabre Pharma Srl, Milan, Italy), a BRM composed of bacterial ribosomal fractions and membrane proteoglycans. Compared with normal controls (NC), before treatment BAL in patients with CB contained increased proportions of neutrophils (NC, 0.8 +/- 0.2 percent; CB, 3 +/- 1 percent), of eosinophils (NC, 0.1 +/- 0.02 percent; CB, 0.6 +/- 0.2 percent); and of lymphocytes (NC, 6 +/- 1 percent; CB, 13 +/- 2 percent; p < 0.01 each comparison) with higher percentages of CD3+ and CD8+ lymphocytes (p < 0.01 each comparison). In BAL from patients with CB there were also higher levels of albumin and of the ratio IgG/albumin (p < 0.01 and p < 0.05, respectively, compared with NC). After Immucytal treatment, the proportions of lymphocytes in BAL in patients with CB were decreased (13 +/- 2 percent before, 6 +/- 1 percent after; p < 0.01). In addition, the posttreatment BAL samples contained significantly fewer neutrophils per milliliter of BAL (3.7 +/- 0.8 x 10(3) neutrophils per milliliter of BAL before, 1.5 +/- 0.5 x 10(3) neutrophils per milliliter after; p < 0.05). No differences were seen for the proportions of lymphocyte subpopulations and for the protein levels between the BAL obtained before and after Immucytal treatment. These data demonstrate the presence of a lower respiratory tract inflammation in patients with CB and suggest that treatment of patients with CB with a BRM may change the proportions of inflammatory cells present in BAL.

The processes of phagocytosis and intracellular killing of bacteria by alveolar macrophages (AMs) and polymorphonuclear leukocytes (PMNs) result in the production of reactive oxygen species that can induce self-damage to the phagocytic cells. N-Acetylcysteine (NAC), a mucolytic agent used to treat chronic respiratory inflammatory disorders, possesses antioxidant properties and has therefore been used for the prevention of damage induced by oxygen radicals. This study was designed to evaluate whether NAC can interfere with the processes of phagocytosis and intracellular killing of bacteria and protect the phagocytic cells from self-killing. Human AM, obtained by bronchoalveolar lavage, and peripheral blood PMNs were cultured with Staphylococcus aureus (American Type Culture Collection 25923 strain) in the presence of different concentrations of NAC (1, 10, and 100 mg/L) and two chromophores (4',6'-diamidino-2-phenylindole dihydrochloride and propidium iodide), which identify live or dead bacteria and dead phagocytes. As compared with PMNs, AMs were more effective in ingesting bacteria (p < 0.05) and were equally effective as intracellular killers (p > 0.05), but were susceptible to a significantly higher rate of self-killing (p < 0.01). The presence of NAC in the cell cultures at the highest dose tested (100 mg/L) induced a significant enhancement of the bactericidal activity of both AMs (p < 0.05) and PMNs (p < 0.05). This increased intracellular killing was not associated with increased proportions of dead phagocytes either in AMs or PMNs cultures (p > 0.05, each comparison), suggesting a protective effect of NAC on damage induced by toxic products generated during phagocytosis.

Pulmonary complications are the most frequent cause of morbidity and mortality in immunocompromised patients. The speed of clinical assessment and the initiation of appropriate therapy is critically related to survival. Fibreoptic bronchoscopy and bronchoalveolar lavage (BAL) had proved useful in making the diagnosis of pulmonary complications in a high proportion of immunocompromised patients, where less invasive techniques, such as blood cultures or sputum induction, have failed to establish a diagnosis. Bronchoscopy and BAL cause little discomfort and low morbidity, and should be performed as early in the disease course as possible, preferably before the onset of respiratory failure.

T-lymphocytes with T-cell antigen receptor (TCR) comprising a gamma chain and a delta chain (gamma delta+ T cells) are believed to be involved in the immune reaction to mycobacterial antigens, and they have been found in increased proportions in sarcoid patients. We evaluated the proportions of gamma delta+ T-lymphocytes and of two major gamma delta+ subpopulations, the V delta 1+ and the V delta 2+ T-cell subsets, in 10 normal blood donors, in 15 patients with tuberculosis (TB), seven of whom had pleural effusion (PE), and in 12 patients with pulmonary sarcoidosis (PS), nine of whom underwent bronchoalveolar lavage (BAL). T-cell subsets were evaluated in peripheral blood (PBL) of all subjects and in PE from patients with TB and in BAL from patients with PS. Compared with normal blood donors, patients with TB had increased proportions of PBL CD3+ gamma delta+ T cells (6 +/- 1% versus 14 +/- 3% of CD3+ T cells, p < 0.05) because of the presence of four patients who had an increase (respectively, 18.3, 22.0, 24.2, and 35.4% of CD3+ T cells) of gamma delta+ T cells. In patients with TB and PE, gamma delta+ T cells were 7.9 +/- 2.7%, a value not different from that in the tubercular PBL and in normal PBL. Although patients with PS had proportions of PBL gamma delta+ T cells (9.2 +/- 3.4%) similar to those in normal PBL, two patients had increased (35 and 31%) PBL gamma delta+ T-lymphocytes.(ABSTRACT TRUNCATED AT 250 WORDS)