Ligation of CD45 and the isoforms CD45RA and CD45RB accelerates the rate of constitutive apoptosis in human eosinophils

M G Blaylock, D W Sexton, G M Walsh

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Background: Eosinophils are important effector cells in asthma pathogenesis, and an understanding of the mechanisms involved in eosinophil apoptosis induction might thus be relevant to the resolution of asthmatic inflammation.

Objective: Our aim was to determine the role of the common leukocyte antigen CD45 and the isoforms CD45RA, CD45RB, and CD45RO in human eosinophil apoptosis induction.

Methods: Immmunostaining and flow cytometry were used to assess CD45 and CD45 isoform expression by cosinophils purified with use of density gradients and immunomagnetic negative selection. Apoptosis induction was measured by binding of fluorescein isothiocyanate-labeled annexin V to eosinophils cultured for 20 hours alone or with saturating quantities of m-lb against CD45. CD45RA, CD45RB, CD45RO, CD9, CD11b, and isotype-matched controls in the presence or absence of GM-CSF.

Results: Freshly isolated eosinophils had high expression of CD45 and CD45RO, modest expression of CD45RB, and low expression of CD45RA. Eosinophils cultured alone for 20 hours were found to be approximately 20% to 25% apoptotic, Incubation with mAb against CD45, CD45RA, and CD45RB resulted in significant (P < .005) enhancement (>100%) of their constitutive rate of apoptosis. Incubation with CD45RO, CD11b CD9 mAb, or isotype controls had no significant effect on the rate of eosinophil constitutive apoptosis, The addition of GM-CSF inhibited eosinophil apoptosis but did not prevent CD15, CD45RA or CD45RB mAb-dependent apoptosis induction.

Conclusion: These data indicate that ligation of CD45. CD45RA, or CD45RB represents a novel pathway for the induction of apoptosis in human eosinophils.

Original languageEnglish
Pages (from-to)1244-1250
Number of pages7
JournalJournal of Allergy and Clinical Immunology
Volume104
Publication statusPublished - 1999

Keywords

  • eosinophils
  • apoptosis
  • CD45
  • allergy asthma
  • cell surface molecules
  • COLONY-STIMULATING FACTOR
  • PROGRAMMED CELL-DEATH
  • CROSS-LINKING
  • TYROSINE PHOSPHATASE
  • REGULATES APOPTOSIS
  • IL-5
  • HEMATOPOIETINS
  • PHAGOCYTOSIS
  • MACROPHAGES
  • NEUTROPHILS

Cite this

Ligation of CD45 and the isoforms CD45RA and CD45RB accelerates the rate of constitutive apoptosis in human eosinophils. / Blaylock, M G ; Sexton, D W ; Walsh, G M .

In: Journal of Allergy and Clinical Immunology, Vol. 104, 1999, p. 1244-1250.

Research output: Contribution to journalArticle

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abstract = "Background: Eosinophils are important effector cells in asthma pathogenesis, and an understanding of the mechanisms involved in eosinophil apoptosis induction might thus be relevant to the resolution of asthmatic inflammation.Objective: Our aim was to determine the role of the common leukocyte antigen CD45 and the isoforms CD45RA, CD45RB, and CD45RO in human eosinophil apoptosis induction.Methods: Immmunostaining and flow cytometry were used to assess CD45 and CD45 isoform expression by cosinophils purified with use of density gradients and immunomagnetic negative selection. Apoptosis induction was measured by binding of fluorescein isothiocyanate-labeled annexin V to eosinophils cultured for 20 hours alone or with saturating quantities of m-lb against CD45. CD45RA, CD45RB, CD45RO, CD9, CD11b, and isotype-matched controls in the presence or absence of GM-CSF.Results: Freshly isolated eosinophils had high expression of CD45 and CD45RO, modest expression of CD45RB, and low expression of CD45RA. Eosinophils cultured alone for 20 hours were found to be approximately 20{\%} to 25{\%} apoptotic, Incubation with mAb against CD45, CD45RA, and CD45RB resulted in significant (P < .005) enhancement (>100{\%}) of their constitutive rate of apoptosis. Incubation with CD45RO, CD11b CD9 mAb, or isotype controls had no significant effect on the rate of eosinophil constitutive apoptosis, The addition of GM-CSF inhibited eosinophil apoptosis but did not prevent CD15, CD45RA or CD45RB mAb-dependent apoptosis induction.Conclusion: These data indicate that ligation of CD45. CD45RA, or CD45RB represents a novel pathway for the induction of apoptosis in human eosinophils.",
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TY - JOUR

T1 - Ligation of CD45 and the isoforms CD45RA and CD45RB accelerates the rate of constitutive apoptosis in human eosinophils

AU - Blaylock, M G

AU - Sexton, D W

AU - Walsh, G M

PY - 1999

Y1 - 1999

N2 - Background: Eosinophils are important effector cells in asthma pathogenesis, and an understanding of the mechanisms involved in eosinophil apoptosis induction might thus be relevant to the resolution of asthmatic inflammation.Objective: Our aim was to determine the role of the common leukocyte antigen CD45 and the isoforms CD45RA, CD45RB, and CD45RO in human eosinophil apoptosis induction.Methods: Immmunostaining and flow cytometry were used to assess CD45 and CD45 isoform expression by cosinophils purified with use of density gradients and immunomagnetic negative selection. Apoptosis induction was measured by binding of fluorescein isothiocyanate-labeled annexin V to eosinophils cultured for 20 hours alone or with saturating quantities of m-lb against CD45. CD45RA, CD45RB, CD45RO, CD9, CD11b, and isotype-matched controls in the presence or absence of GM-CSF.Results: Freshly isolated eosinophils had high expression of CD45 and CD45RO, modest expression of CD45RB, and low expression of CD45RA. Eosinophils cultured alone for 20 hours were found to be approximately 20% to 25% apoptotic, Incubation with mAb against CD45, CD45RA, and CD45RB resulted in significant (P < .005) enhancement (>100%) of their constitutive rate of apoptosis. Incubation with CD45RO, CD11b CD9 mAb, or isotype controls had no significant effect on the rate of eosinophil constitutive apoptosis, The addition of GM-CSF inhibited eosinophil apoptosis but did not prevent CD15, CD45RA or CD45RB mAb-dependent apoptosis induction.Conclusion: These data indicate that ligation of CD45. CD45RA, or CD45RB represents a novel pathway for the induction of apoptosis in human eosinophils.

AB - Background: Eosinophils are important effector cells in asthma pathogenesis, and an understanding of the mechanisms involved in eosinophil apoptosis induction might thus be relevant to the resolution of asthmatic inflammation.Objective: Our aim was to determine the role of the common leukocyte antigen CD45 and the isoforms CD45RA, CD45RB, and CD45RO in human eosinophil apoptosis induction.Methods: Immmunostaining and flow cytometry were used to assess CD45 and CD45 isoform expression by cosinophils purified with use of density gradients and immunomagnetic negative selection. Apoptosis induction was measured by binding of fluorescein isothiocyanate-labeled annexin V to eosinophils cultured for 20 hours alone or with saturating quantities of m-lb against CD45. CD45RA, CD45RB, CD45RO, CD9, CD11b, and isotype-matched controls in the presence or absence of GM-CSF.Results: Freshly isolated eosinophils had high expression of CD45 and CD45RO, modest expression of CD45RB, and low expression of CD45RA. Eosinophils cultured alone for 20 hours were found to be approximately 20% to 25% apoptotic, Incubation with mAb against CD45, CD45RA, and CD45RB resulted in significant (P < .005) enhancement (>100%) of their constitutive rate of apoptosis. Incubation with CD45RO, CD11b CD9 mAb, or isotype controls had no significant effect on the rate of eosinophil constitutive apoptosis, The addition of GM-CSF inhibited eosinophil apoptosis but did not prevent CD15, CD45RA or CD45RB mAb-dependent apoptosis induction.Conclusion: These data indicate that ligation of CD45. CD45RA, or CD45RB represents a novel pathway for the induction of apoptosis in human eosinophils.

KW - eosinophils

KW - apoptosis

KW - CD45

KW - allergy asthma

KW - cell surface molecules

KW - COLONY-STIMULATING FACTOR

KW - PROGRAMMED CELL-DEATH

KW - CROSS-LINKING

KW - TYROSINE PHOSPHATASE

KW - REGULATES APOPTOSIS

KW - IL-5

KW - HEMATOPOIETINS

KW - PHAGOCYTOSIS

KW - MACROPHAGES

KW - NEUTROPHILS

M3 - Article

VL - 104

SP - 1244

EP - 1250

JO - Journal of Allergy and Clinical Immunology

JF - Journal of Allergy and Clinical Immunology

SN - 0091-6749

ER -