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Our data confirm that CD4 and CD8 T cells isolated from BAL wash fluid coexpress α4β1 at a high level, supporting the use of these markers as a measure of lung homing capacity.
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At 8 days postinfection animals were euthanized and BAL washes were collected for determination of anti-Fim2 IgG (a) and IgA (b) antibodies.
The cell pellet from a 10 ml aliquot of BAL was washed, counted and resuspended at a concentration of 0.02 x 10 cells.ml-1.
CCL3 protein concentration was measured in the sera, nasal wash, BAL and lung homogenate supernatant.
Therefore, current diagnostic approaches include non-culture/histopathologic modalities including combination of chest CT imaging and non-invasive, non-culture based tools such as galactomannan (GM) antigen detection in bronchoalveolar lavage (BAL) and bronchial wash (BW) [ 7– 11].
Pooled BAL cells were washed in Ca+2+Mg+2-free PBS by alternate centrifugation (800×g for 10 min at 4°C) and resuspension.
BAL cells were washed twice with ice-cold culture medium, and cell viability and concentration were determined by staining with trypan blue (0.04%).
BAL cells were washed with PBS.
Pooled BAL cells were washed in Ca2+/Mg2+-free PBS by alternate centrifugation (200 g, 10 min, 4 °C).
For immunofluorescence staining, 10 BAL cells were washed once with phosphate-buffered saline (PBS), and resuspended in 50 μl of PBS containing 2% fetal bovine serum (FBS), 0.05% sodium azide and 5 μg/ml mAbs.
Fifteen volunteers all underwent bronchoscopies with bronchial wash and BAL on three occasions, each being 3 h after inhalation provocation.
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