Abstract
Mixed lymphocyte reactions (MLRs) remain central to the characterization of cellular allo-interactions. Here we show that irradiation, as used to ‘silence’ a given cell-population in unidirectional (‘one-way’) MLRs, is unable to abolish cytokine-production even at doses much higher than usually applied. By contrast, using target cells silenced via a formaldehyde-based fixation-protocol, we demonstrate feasibility to detect – in a true one-way reaction – secretion of IFNγ by alloreactive NK cells. This simple, fixation-based protocol provides an accurate, robust and time-efficient means for assessing alloreactivity, avoiding cytokine-production by the MLR stimulator cells.
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