An improved protocol for measuring cytotoxic T cell activity in anatomic compartments with low cell numbers

The study of target cell lysis and cytokine production are valuable tools t o characterize antigen-specific T and NK cell function during virus infecti ons. After localized infections in compartments such as the lung or the bra in, however, cell numbers isolated from these organs are too low to perform standard assays with individual mice. Here, we report a few simple modific ations of the classical Cr-51 release assay allowing reduction of the numbe r of required effector cells by a factor of 10 without loosing sensitivity or specificity. Using not more than 4 X 10(5) effector cells, we were able to study ex vivo virus-specific CTL or NK activity from the lungs of indivi dual mice after infection with respiratory syncytial virus (RSV) and from t he brains of mice infected with Boma disease virus (BDV). Flow cytometric a nalysis of interferon-gamma production by virus-specific T cells including appropriate controls was achieved with as few as 105 effector cells. (C) 20 01 Elsevier Science B.V. All rights reserved.