Although the role of L-carnitine (L-Cn) as a cofactor in the oxidation of long-chain fatty acids has been well established, this agent has also been recognized to have an important role in the regulation of carbohydrate metabolism, and consequently, the maintenance of cell membrane structure and cell viability. L-Cn has been reported to reduce the apoptotic levels of CD4+ and CD8+ cells. It has also been demonstrated to interfere with cells of the monocytic lineage by regulating their ability to produce growth factors that ultimately affect both T and B lymphocytic subsets. Therefore, in this study, we examined whether this agent affects the antigenic response of immune cells and determined the relative numbers of immune cells in the murine spleen after in vitro and in vivo treatment. The results showed that L-Cn reduces the relative numbers of CD8+, CD4+ and Ly5+ cells. This observation was consistent in all systems studied including (a) in vitro inoculation of antigen (DNP-HSA) and L-Cn, (b) in vitro priming of spleen cells treated with L-Cn in vivo, and (c) in vivo immunization and L-Cn administration. In all cases, the reduction of T lymphocytes correlated with the decreased production of interleukin-2. L-Cn, however, did not affect the production of specific antibody, which indicates that the observed reduction of Ly5-positive cells is due to cell differentiation of B cells to plasma cells.