We investigated the in vivo role of interleukin 5 (IL-5) and its receptor (IL-5R) in eosinophil growth and differentiation. When mice were administered IL-5 i.p., an increase in the number of eosinophils was observed within 5 days in peripheral blood and the peritoneal cavity. Hypereosinophilia was observed in IL-5 transgenic mice who displayed constitutive production of IL-5. A binding assay with 35S-labeled IL-5 revealed the presence of two classes of IL-5 binding sites (low and high affinity) on the surface of eosinophils. IL-5Rs on eosinophils were recognized using mAbs against murine IL-5R. When the IL-5 transgenic mice were passively administered with anti-murine IL-5R mAbs, the number of recognizable eosinophils in peripheral blood dropped within 5 days to normal levels. The antibody treatment also prevented the increase in the number of eosinophils in IL-5-injected mice. The inhibition of the above experimental eosinophilia was also observed by the passive administration of anti-IL-5 mAb. The results of the in vivo experiments clearly demonstrate that IL-5 plays an essential role in in vivo eosinophilopoiesis and may be acting on eosinophils or their precursors directly through IL-5Rs, resulting in preferential growth of this lineage of hematopoietic cells. It can also be stressed that IL-5 regulates a specific lineage of hematopoietic cells (eosinophils).