The dependence of human lymphocyte functions on the exogenous provision of glutamine (GLN) was evaluated in a series of in vitro experiments. The transcription of early activation markers (IL-2, IL-2-receptor, IL-4, IL-4, GM-CSF, IFN gamma) as evaluated by polymerase chain reaction was observed even in the absence of exogenous GLN. In contrast, later events of lymphocyte activation including cytokine production, proliferation of lymphocytes and lymphokine-activated killer cell activity were found to depend on exogenous GLN provision. These in vitro results are discussed in the context of established data on the reduction of peripheral blood GLN concentrations in critically ill patients and in view of recent studies reporting improved outcome of critically ill patients following GLN substitution. By and large, these data support the concept of GLN substitution in critical illness. However, the definition of indications and dose-response relationships clearly require further clinical studies.