title: Release of granule proteins from eosinophils cultured with IL-5.
abstract: Eosinophils isolated from normal individuals were cultured in the presence of human rIL-5 (hrIL-5) for up to 14 days, and the effects of this exposure were determined. First, the hrIL-5-cultured eosinophils were activated and degranulated more readily than freshly isolated eosinophils. For example, eosinophils cultured for 7 days with hrIL-5 released 30 and 10% of granule eosinophil-derived neurotoxin (EDN) when exposed to Sepharose 4B beads coupled to secretory IgA and IgG, respectively, whereas freshly isolated eosinophils released only 19 and 4%, respectively, of their EDN in response to the same stimuli. Degranulation of hrIL-5-cultured eosinophils was not augmented by further exposure to hrIL-5, whereas degranulation of freshly isolated cells to secretory IgA and IgG beads was increased by exposure to hrIL-5. Second, eosinophils cultured with hrIL-5 had prolonged viability in vitro. For example, after four days of culture with 50 U/ml of hrIL-5, 86% of eosinophils were viable compared to 12% in medium alone. Third, hrIL-5-cultured eosinophils became hypodense, and electron microscopy showed that they contained granules with core and matrix lucency and with evidence of granule fusion. Fourth, hrIL-5-cultured eosinophils spontaneously lost 30 to 60% of their EDN, eosinophil cationic protein, and eosinophil peroxidase and about 50% of their eosinophil granule major basic protein content compared to freshly isolated eosinophils, and all four of the granule proteins were released into the culture medium. Fifth, detailed studies of eosinophils cultured in hrIL-5 showed that 89 +/- 10% of the starting quantity of EDN could be recovered at 7 days. Whereas 99 +/- 1% of the EDN at day 0 was cell associated, by 7 days 60 +/- 9% was in the cell supernatants. Thus, hrIL-5 activates eosinophils, increases their viability, decreases their density, and their content of granule proteins and causes release of the granule proteins into culture fluids. The striking loss of granule proteins during culture with hrIL-5 may be an important mechanism for deposition of these cationic toxins in various diseases where IL-5 plays a role.
authors.affiliation: Department of Immunology, Mayo Clinic, Rochester, MN 55905.
authors.forename: H
authors.initials: H
authors.lastname: Kita
authors.forename: D A
authors.initials: DA
authors.lastname: Weiler
authors.forename: R
authors.initials: R
authors.lastname: Abu-Ghazaleh
authors.forename: C J
authors.initials: CJ
authors.lastname: Sanderson
authors.forename: G J
authors.initials: GJ
authors.lastname: Gleich
mesh_terms: D001798:Blood Proteins / Q000032:analysis / Q000378:metabolism*; D015550:Cell Degranulation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D047091:Eosinophil Granule Proteins; D047208:Eosinophil-Derived Neurotoxin; D004804:Eosinophils / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007136:Immunoglobulins / Q000502:physiology; D015848:Interleukin-5 / Q000494:pharmacology*; D009498:Neurotoxins / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D012260:Ribonucleases
chemical_list: D001798:Blood Proteins; D047091:Eosinophil Granule Proteins; D007136:Immunoglobulins; D015848:Interleukin-5; D009498:Neurotoxins; D011994:Recombinant Proteins; D047208:Eosinophil-Derived Neurotoxin; D012260:Ribonucleases