Determination of the number of subtypes of lymphocytes present in bronchoalveolar lavage (BAL) by FACS.

BALs were pooled out from three mice treated with adenoviral gene vector expressing murine granulocyte-macrophage colony-stimulating factor (AdGM-CSF) or Addl70-3, respectively, 7 days after gene transfer. Cells were immunostained with different combinations of monoclonal antibodies and examined by FACS analysis. Then, the absolute cell numbers of CD4, CD8, and natural killer cells were calculated, based on the total cell recovery and the percentage of each subtype.