multiple, large areas of airway tissue. Often it is not feasible to quantify stained area and staining intensity in a standardized way to include all available areas in the analysis. Second, it is not to obtain robust estimates of inflammatory cell counts and cytokine always possible to stain all microscopic slides in one batch, expression from multiple large areas of histopathologic sections. especially in large studies. Third, there seems to be an in- pers comparison between observers. Therefore, to achieve interferon-␥ protein was performed on frozen tissue sections, using minimal procedural variability the assessments in biopsy speci-3-amino-9-ethylcarbazole as chromogen and hematoxylin as counmens should preferably be done in random order and within terstaining. IL-4 and IL-5 messenger RNAs were localized by in situ a reasonable time span by one observer within a single study. Obviously, this is not always possible in practice. Automated image analysis might deal with these issues by we developed a fully automated inflammatory cell-counting image analysis with interactive digital cell counting and semiInflammatory cell number and cytokine expression in bronquantitative scoring of cytokine expression in bronchial biopsy chial biopsies are increasingly considered to be a major outspecimens in terms of repeatability, and agreement in a crosscome in asthma and chronic obstructive pulmonary disease sectional and longitudinal design. (COPD) research (1). However, the inflammatory infiltrate often shows a patchy focal pattern, sometimes with clusters METHODS in which the cells are difficult to count (2). Therefore, there Subjects might be considerable within-section, within-biopsy, and between-biopsy variability ("true" variability). To achieve satisThis study used bronchial biopsy specimens from patients with atopic asthma who participated in two different studies, and was performed factory stable inflammatory cell counts in bronchial biopsy to examine automated assessment of cytokine expression (27 patients) specimens it is recommended that multiple large areas of (6) and inflammatory cell counts (25 patients) (7). All patients (21 males airway tissue be examined (3-5). and 31 females) were nonsmokers at the time of recruitment (nonsmokers In addition to this true variability, the number of inflammafor more than 1 year, smokers for fewer than 5 pack-years), were atopic, tory cells that is determined will also be influenced by procewere between 18 and 50 years of age, and had a history of episodic dural factors. First, it is labor-intensive to assess cell counts in chest tightness and wheezing in the previous year. Atopy was assessed through a positive skin prick test reaction (wheals greater than 3 mm in diameter) to one or more common airborne allergen extracts (Soluprick; ALK, Copenhagen, Denmark). Prebronchodilator forced expired volume (Received in original form May 2, 2002; accepted in final form January 29, 2003) in 1 second (FEV 1 ) was more than 50% of the predicted value and more than 1.5 L, whereas postbronchodilator FEV 1 was within the normal range (more than 80% predicted). At the time of patients' entry into
Sont et al. REF assessed inflammatory cell counts and cytokine expression in immunostained sections of bronchial tissue.
Fully Automated Assessment of Inflammatory Cell Counts and Cytokine Expression in Bronchial Tissue
American journal of respiratory and critical care medicine
American journal of respiratory and critical care medicine
Medicine