Release of granule proteins from eosinophils cultured with IL-5

Hirohito Kita, Deborah A. Weiler, Randa Abu-Ghazaleh, Colin J. Sanderson, Gerald J. Gleich

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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.

Original languageEnglish (US)
Pages (from-to)629-635
Number of pages7
JournalJournal of Immunology
Volume149
Issue number2
StatePublished - Jul 15 1992

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Eosinophil Granule Proteins
Interleukin-5
Eosinophils
Eosinophil-Derived Neurotoxin
Secretory Immunoglobulin A
Proteins
Eosinophil Major Basic Protein
Immunoglobulin G
Eosinophil Peroxidase
Eosinophil Cationic Protein

ASJC Scopus subject areas

  • Immunology

Cite this

Kita, H., Weiler, D. A., Abu-Ghazaleh, R., Sanderson, C. J., & Gleich, G. J. (1992). Release of granule proteins from eosinophils cultured with IL-5. Journal of Immunology, 149(2), 629-635.

Release of granule proteins from eosinophils cultured with IL-5. / Kita, Hirohito; Weiler, Deborah A.; Abu-Ghazaleh, Randa; Sanderson, Colin J.; Gleich, Gerald J.

In: Journal of Immunology, Vol. 149, No. 2, 15.07.1992, p. 629-635.

Research output: Contribution to journalArticle

Kita, H, Weiler, DA, Abu-Ghazaleh, R, Sanderson, CJ & Gleich, GJ 1992, 'Release of granule proteins from eosinophils cultured with IL-5', Journal of Immunology, vol. 149, no. 2, pp. 629-635.
Kita H, Weiler DA, Abu-Ghazaleh R, Sanderson CJ, Gleich GJ. Release of granule proteins from eosinophils cultured with IL-5. Journal of Immunology. 1992 Jul 15;149(2):629-635.
Kita, Hirohito ; Weiler, Deborah A. ; Abu-Ghazaleh, Randa ; Sanderson, Colin J. ; Gleich, Gerald J. / Release of granule proteins from eosinophils cultured with IL-5. In: Journal of Immunology. 1992 ; Vol. 149, No. 2. pp. 629-635.
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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.",
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N2 - 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.

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