We have used immunohistochemistry and monoclonal antibodies to analyze the phenotypic composition and activation status of the cellular infiltrate of bronchial biopsies obtained by fiber optic bronchoscopy of 11 atopic asthmatic subjects (FEV1 % predicted range 78 to 114), 9 atopic nonasthmatic control subjects, and 10 normal healthy subjects. Examination of mucosal biopsies obtained from both central (level I) and subsegmental (level II) bronchi showed that the highest number of CD45-, DC3-, DC4-, and CD8-positive cells were found in the group with asthma. There was a significant increase in the number of interleukin-2 receptor (CD25)-positive cells (a marker of lymphocyte activation) at airway level I in the asthmatic group compared with both nonasthmatic atopic (p < 0.05) and normal control subjects (p < 0.01). Eosinophil numbers were significantly increased in asthma at both airway levels and at airway level II in the nonasthmatic atopic group when compared with normal healthy control subjects (p < 0.05). EG2-positive cells (an index of secretion of eosinophil cationic protein following activation) were found at both airway levels in the asthmatic group and at level I in the nonasthmatic atopic control group (p < 0.05). When asthmatic subjects were compared with normal healthy subjects, there was a reduction in the number of neutrophil elastase-positive cells in the asthmatic subjects which, as a percentage of leukocytes, was significant (p = 0.05). When normoresponsive subjects were compared with hyperresponsive subjects (PC20 < 8 mg/ml methacholine), the hyperresponsive subjects had a significantly higher percentage of CD25-positive cells at airway level I (p < 0.01) and of EG2-positive cells at levels I and II (p < 0.01). These results support the view that the recruitment and “activation” of lymphocytes and eosinophils are important in the pathogenesis of bronchial asthma.

We have attempted to identify mRNA for IL-5 in endobronchial mucosal biopsies from asthmatics and controls, using the technique of in situ hybridization. Bronchial biopsies were obtained from 10 asthmatics and 9 nonatopic normal controls. A radio-labeled cRNA probe was prepared from an IL-5 cDNA and hybridized to permeabilized sections. These were washed extensively before processing for autoradiography. An IL-5-producing T cell clone derived from a patient with the hyper-IgE syndrome was used as a positive control. As a negative control, sections were also treated with a "sense" IL-5 probe. Specific hybridization signals for IL-5 mRNA were demonstrated within the bronchial mucosa in 6 out of the 10 asthmatic subjects. Cells exhibiting hybridization signals were located beneath the epithelial basement membrane. In contrast, there was no hybridization in the control group. No hybridization was observed with the sense probe. The six IL-5 mRNA-positive asthmatics tended to have more severe disease than the negative asthmatics, as assessed by symptoms and lung function, and showed a significant increase in the degree of infiltration of the bronchial mucosa by secreting (EG2+) eosinophils and activated (CD25+) T lymphocytes. Within the subjects who showed positive IL-5 mRNA, there was a correlation between IL-5 mRNA expression and the number of CD25+ and EG2+ cells and total eosinophil count. This study provides evidence for the cellular localization of IL-5 mRNA in the bronchial mucosa of asthmatics and supports the concept that this cytokine regulates eosinophil function in bronchial asthma.

With the use of monoclonal antibodies (mAb) and immunohistology, the numbers of phenotypically distinct cells infiltrating lung tissue from 15 postmortem (PM) cases of fatal asthma were quantified and compared with 6 cases of cystic fibrosis (CF) (three postmortem, three transplant) and 10 nonasthmatic cases of sudden death matched for age and sex. Tissue eosinophilia was significantly greater (p < 0.001) in the fatal asthma group than in the CF or sudden death controls. In asthma, approximately 40% of the eosinophilic infiltrate was EG2 positive (an indication of eosinophil activation and secretion of eosinophil cationic protein). The numbers of eosinophils and EG2 positive cells were significantly elevated in the subjects with acute severe asthma who had had a duration of terminal illness exceeding, as compared with less than, 24 h (p < 0.05). When compared with the sudden death controls, there were increases in the numbers of Dako L C positive cells (i.e., CD45 positive “total leukocytes”) in both fatal asthma and CF (p < 0.01 and 0.05, respectively). The mean number of MT-1 positive (T) cells in the asthma and CF groups was approximately twice that of the control (p < 0.05 and 0.01, respectively). The mean number of MB2 positive (B) cells was similar for both the asthma and sudden death control groups but was significantly increased in CF (p < 0.05). The average T:B cell ratios were 6:1, 1:1, and 2:1 in the fatal asthma, CF, and control groups, respectively. The results support a role for the T lymphocyte in the pathogenesis of fatal asthma and CF. However, the high numbers of B cells and the low numbers of eosinophils in CF distinguish CF from asthma. Asthma in contrast is characterized by a high T:B cell ratio and the presence in airway tissue of relatively large numbers of “activated” eosinophils.

Journal of Medical Internet Research - International Scientific Journal for Medical Research, Information and Communication on the Internet #Preprint #PeerReviewMe: Warning: This is a unreviewed preprint. Readers are warned that the document has not been peer-reviewed by expert/patient reviewers or an academic editor, may contain misleading claims, and is likely to undergo changes before final publication, if accepted, or may have been rejected/withdrawn. Readers with interest and expertise are encouraged to sign up as peer-reviewer, if the paper is within an open peer-review period. Please cite this preprint only for review purposes or for grant applications and CVs (if you are the author). Background: Cardiac rehabilitation (CR) is an essential part of coronary heart disease (CHD) management. However, patients exiting a center-based CR program have difficulty retaining its benefits. Objective: The purpose of the TRiCH study is to evaluate the added benefit of a home-based (HB) CR program with telemonitoring guidance on physical fitness in patients with coronary artery disease (CAD) who are being discharged from a phase II ambulatory CR program, and to compare its effectiveness to a prolonged center-based (CB) CR intervention. Methods: Between February 2014 and August 2016, 90 CAD patients (61.2±7.6yrs, 89% males, 1.73±0.7m, 82.9±13kg, 27.5±3.4kg/m2) who successfully completed a three month ambulatory CR program were randomly allocated to one of three groups: HB (=30), CB (=30) or a control group (CG) (=30) on a 1:1:1 basis. HB patients received a home-based exercise intervention with telemonitoring guidance consisting of weekly emails or phone calls, CB patients continued the ambulatory-hospital CR and CG patients received usual care including the recommendation to remain physically active. All patients underwent cardiopulmonary exercise testing to assess peak oxygen uptake (VO2P) at baseline and after the 12 week intervention period. Secondary outcomes included physical activity behavior, anthropometric characteristics, cardiovascular risk factors and quality of life. Results: The increase in VO2P was larger following 12 weeks of intervention in the CB group (P = .03) and HB group (P = .04) compared to the control group. In addition, oxygen uptake at the first ventilatory threshold (P-interaction = .03) and the second ventilatory threshold (P-interaction = .03) increased significantly more in the HB group compared to CB. No significant changes were found in the secondary outcomes. Conclusions: Adding a HB exercise program with telemonitoring guidance following completion of a phase II ambulatory CR program results in further improvement of physical fitness and is as equally effective as prolonging a CB CR in patients with coronary artery disease. Clinical Trial: NCT02047942.

The kinetics and phenotype of T lymphocytes infiltrating the airways of guinea pigs undergoing late-phase asthmatic reactions (LAR) were studied with monoclonal antibodies, cytofluorimetry, and immunocytochemistry. Challenge of sensitized animals with aerosolized ovalbumin was followed by early (2 h) and late-phase (17 h) bronchoconstriction. The induction of hypersensitivity, by aerosolized antigen, was associated with an increase in mucosal T cell numbers, which consisted almost entirely of CD8+ T cells. Following allergen challenge of fully sensitized animals, a biphasic rise in total T cell (CD3+) numbers was observed in the bronchial mucosa, peaking at 17 and 48 h. A similar pattern of T cell accumulation was observed in the bronchial adventitia but with an extra early peak at 2 h. In contrast to the T cell influx of the sensitization phase, the postchallenge infiltrate consisted largely of CD3 + ,CD8− cells. Eosinophil numbers were elevated in both submucosa and adventitia, with a single broad peak between 17 and 48 h. T cell infiltration was compared with eosinophil accumulation: while correlations between T cell and eosinophil numbers varied over the 96 h of the experiment, strong associations were observed between CD8+ numbers and eosinophils in the adventitia at 6 h (r = 0.733, p < 0.01) and between CD3+ numbers and eosinophils in the submucosa at 72 h (r = 0.88, p < 0.001). No significant changes were detected in T cell or eosinophil numbers in the lung parenchyma. There was a postchallenge increase in eosinophils (but not T cells) in bronchoalveolar lavage (BAL). In contrast, analysis of blood leukocytes showed no changes in T cell total or subset numbers during the progression of the LAR. These results indicate that accumulation of T cells in the airways is a feature of the LAR in this model. These observations are consistent with human BAL and skin studies of the allergen-induced late-phase reaction. These data also illustrate that evaluation of distant compartments (blood and BAL) may not fully reflect rapidly evolving tissue infiltration in the bronchial mucosa.