Since granulocyte colony-stimulating factor (G-CSF) is thought to be a granulocyte lineage-specific cytokine, G-CSF receptors on blood cells other than those of granulocyte or monocyte lineage have not been well investigated. We now report that G-CSF receptors are present on platelets. The expression of G-CSF receptors on platelets was demonstrated by flow cytometry and radioreceptor assay. The mean number of G-CSF-binding sites per cell was 41 and the binding affinity was high (Kd 300 pM), similar to the affinity observed on granulocytes. Cross-linking assay revealed that G-CSF receptors were present on a single subunit protein of approximately 150 kD on the platelets. To clarify whether or not G-CSF might produce some direct functional influence on platelet response, the effects on platelet aggregation were studied. Although G-CSF itself did not affect platelet aggregation in vitro, preincubation with G-CSF augmented a secondary aggregation of platelets induced by low concentrations of adenosine diphosphate (ADP). There was a dose-response relationship for this G-CSF activity at concentrations of up to 10 ng/ml. Furthermore, the augmented ADP-induced secondary aggregation of platelets on G-CSF receptors was completely abrogated in the presence of anti-G-CSF polyclonal antibodies. These results indicate that platelets possess functional G-CSF receptors.