Interleukin-5 is the predominant eosinophilopoietin produced by cloned T lymphocytes in hypereosinophilic syndrome

Abstract

Cloned T lymphocytes (TLC) of the CD4+CD8- phenotype established from peripheral blood of a patient with idiopathic hypereosinophilic syndrome (HES) were found to release a lineage-specific eosinophilic colony-stimulating factor (Eo-CSF). The present study was undertaken to identify the lymphokine accounting for this Eo-CSF activity. Comparison of TLC-derived Eo-CSF with recombinant human interleukin-5 (rhIL-5), recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and recombinant human interleukin-3 (rhIL-3) by in vitro clonogenic assays revealed similar bioactivity of HES-derived Eo-CSF and IL-5. Neutralization studies using specific antibodies against IL-5, GM-CSF and IL-3 confirmed that IL-5 mainly accounts for the Eo-CSF activity in all 9 HES-derived TLC tested. Eosinophilic colony (CFU-Eo) formation supported by conditioned media of the TLC was significantly inhibited in all clones by addition of anti-IL-5 monoclonal antibody (MAB) to the conditioned media. Inhibition by anti-IL-5 MAB was specific and dose-dependent. In 2 of the 9 clones, anti-GM-CSF antibodies could partially neutralize the Eo-CSF activity in the conditioned media. In 4 clones, addition of a combination of anti-IL-5 MAB and anti-GM-CSF antiserum to the conditioned media reduced CFU-Eo formation significantly more than addition of anti-IL-5 MAB alone. In none of the TLC could a significant role for IL-3 in eosinophilic colony formation be shown. These results were confirmed at the mRNA level. Cytokine transcripts were detected by reverse transcription (RT) and subsequent polymerase chain reaction (PCR). Under the same experimental conditions, all HES-derived TLC, but only one third of tested TLC from healthy donors, expressed IL-5 mRNA 5 days after stimulation. In control TLC with inducible IL-5 mRNA expression, IL-5 transcripts were found for only 3 days after stimulation. In contrast, HES-derived TLC contained IL-5 mRNA at least until day 18 after restimulation. All HES clones expressed GM-CSF mRNA upon stimulation. Two HES-derived TLC were found to lack IL-3 mRNA even after stimulation. Whereas IL-5 was expressed abundantly in all HES-clones, the intensity of PCR products for GM-CSF and IL-3 showed striking differences. Our in vitro results suggest that IL-5 produced by activated CD4+ T lymphocytes plays a crucial role in the induction of eosinophilia in HES. In addition, GM-CSF but not IL-3 seems to contribute partially to the increased eosinophil production in HES.