Patients and Control Subjects
Bronchoalveolar lavage was performed in 100 patients with histologically proven sarcoidosis and 14 control subjects. The characteristics of the patients and control subjects are described in Table 1. The patients were divided into three groups based on their clinical presentation. Apnea
Croup A consisted of patients whose disease was detected on routine chest x-ray film, without symptoms or knowledge of the exact duration and the time of onset of the disease (11 NSm and 7 Sm); group В included those with respiratory and general constitutional symptoms (50 NSm and 10 Sm); and group С included patients with erythema nodosum and/or arthralgia and hilar lymph-adenopathy (ie, M. Lflfgren; 16 NSm and 6 Sm). All patients had stage 1 or II disease and none had stage III disease, as evidenced on x-ray films. The majority of the patients were NSms, 77 of 100 (Table 1). The initial BAL fluid samples of consecutive sarcoidosis patients obtained at die time of the diagnosis, within 2 weeks after admission to our hospital, were used for this study. No patient was receiving corticosteroid or other treatment either at the time of or before the lavage. The control group consisted of individuals without chest abnormalities or a history of pulmonary abnormalities or disease (11 NSm; 3 Sm). The studied groups were subdivided according to their smoking status (Table 1).
Bronchoalveolar lavage was performed as previously reported during fiberoptic bronchoscopy.“ At the same time, blood samples were taken. In short, the procedure was as follows. After premedication with atropine and sometimes diazepam or codeine, and local anesthesia of the larynx and bronchial tree with 0.5 percent tetracaine, BAL was performed by standardized washing of the right middle lobe with four 50-ml aliquots of sterile saline solution (0.9 percent NaCl) at room temperature. Lavage fluid samples, kept on ice in a siliconized specimen trap, were centrifuged (10 min, 350 g) and separated into cells and supernatant. The cell pellet was washed twice, counted, and suspended in minimal essential medium (Gibco, Grand Island, NY), supplemented with 1 percent bovine serum albumin (Organon, Teknika, Box tel, the Netherlands). Preparations of the cell suspension were made in a cytocentrifiige (Shandon). Cytospin slides of BAL cells were stained with May-Griinwald-Giemsa (Merck, Darmstadt, Germany) for cell differentiation. At least 1,000 cells were counted.
If more than 15 percent lymphocytes were present in the BAL fluid samples, T lymphocyte subpopulations were determined. Identification of T lymphocytes reacting with monoclonal antibodies was performed by means of a conventional indirect immunofluorescence technique. Total T lymphocytes and subpopulations were recognized by staining with monoclonal antibodies (CD3, CD4, and CD8). Monoclonal antibodies, CD3 (OKT3), CD4 (OKT4), and CD8 (OKT8), were obtained from Ortho-Pharmaceuticals (Beerse, Belgium) and subsequently labelled with FITC-conjugated goat-antimouse-immunoglobulin (Nordic, Immunological Laboratories, Tilburg, the Netherlands and from the Central Laboratory of the Netherlands Red Cross Blood Transfusion Service [CLB], Amsterdam, the Netherlands). Results were expressed as a percentage of lymphocytes.
To investigate whether there were statistically significant differences between the three categories of sarcoidosis patients, the Kruskal-Wallis one-way analysis of variance test was used. Each category, group A (no symptoms), group В (respiratory and general constitutional symptoms), and group С (erythema nodosum and hilar lymphadenopathy [M. Lofgren]), respectively, denoted the clinical presentation of the patient.
The Mann-Whitney U test, a pairwise comparison, was used to evaluate differences between Sms and NSms in each group, as well as the differences in each category with the control subjects. The probability values less than 0.05 were considered to be significant.
Table 1—Characteristics of the Groups Studied
|Group||No. of Cases||Age, yr*||Sex|
|Control subjects||11||39.4 (27-66)||7||4|
|Control subjects||3||41.0 (30-60)||2||1|