Macrophage migration inhibitory factors (MIFs) of mouse and guinea pig have been thoroughly characterized with regard to molecular weight and isoelectric points. Several molecular weight species have been identified. In a comparative study with purified MIFs it was found that these molecules were distinct from a series of other lymphokines, particularly so from macrophage activating activities. Investigations on the molecular weight heterogeneity of MIF have led us to a transglutaminase-like activity which was found to be expressed in certain subsets of macrophages. The question whether low molecular weight factors are polymerized by this enzyme to oligomers is further investigated. Studies on the induction by lymphokines of interferon and plasminogen activator revealed a great heterogeneity of responding macrophages. In studies on the biological basis of the functional heterogeneity of macrophages, the question was investigated whether the heterogeneity was due to different macrophage subpopulations or to intermediate relatively stable phenotypes on their way to maturity and senescence. To approach this question, the bone marrow liquid culture system was used as a developing system. Our data are summarized in a unifying model which takes into account the different constitutive and inducible functions during the cell cycle. Accordingly, lymphokines may act either as differentiation signals, as mitogens or activating signals.