CD44 and CD69 represent different types of cell-surface activation markers for human eosinophils

Kenji Matsumoto, James Akwa Appiah-Pippim, Robert P. Schleimer, Carol A. Bickel, Lisa A. Beck, Bruce S. Bochner

Research output: Contribution to journalArticle

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Abstract

Eosinophils (EOS) purified from peripheral blood or late-phase bronchoalveolar lavage (BAL) were analyzed with 473 monoclonal antibodies (mAbs) from the Fifth International Workshop on Human Leukocyte Antigens in an attempt to identify markers of EOS activation. Two strategies were used: (1) to look for surface markers absent on fresh EOS but present after in vivo activation (e.g., in late-phase BAL fluid [BALF]) or after in vitro culture for up to 72 h with cytokines (≤ 10 ng/ml of interleukin-3 [IL-3], IL-5, or granulocyte-macrophage colony-stimulating factor [GM-CSF]); and (2) to look for markers constitutively expressed on fresh EOS that were increased after activation in vivo or after culture in vitro. With indirect immunofluorescence and flow cytometry, the first approach revealed that among approximately 350 mAbs tested, only those recognizing CD69 became bound to late-phase BALF EOS or cytokine-cultured EOS, but not to fresh EOS. Using the second approach, we observed statistically significant concentration-and time-dependent increases in CD44 expression in EOS cultured with IL-3, IL-5, or GM-CSF (∼ 2-fold increase in fluorescence intensity, P < 0.05), but not with interferon-γ (IFN-γ) (up to 100 ng/ml), whereas levels of 15 other constitutively expressed markers were unchanged. Despite increased expression, neither fresh nor cytokine-cultured EOS adhered to immobilized hyaluronate, a ligand for CD44. Additionally, simultaneous comparison of hypodense (specific gravity < 1.085 g/liter) and normodense (specific gravity > 1.085 g/liter) EOS from allergic donors consistently revealed higher levels of CD44 expression (∼ 3-to 8-fold) but not CD69 expression on hypodense EOS. We conclude that CD69 and CD44 represent different types of activation markers for human EOS. These findings may be useful in assessing the state of EOS activation in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)860-866
Number of pages7
JournalAmerican journal of respiratory cell and molecular biology
Volume18
Issue number6
DOIs
StatePublished - Jan 1 1998

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Eosinophils
Chemical activation
Granulocyte-Macrophage Colony-Stimulating Factor
Interleukin-3
Interleukin-5
Monoclonal Antibodies
Cytokines
Dimercaprol
Fluids
Flow cytometry
HLA Antigens
Blood
Fluorescence
Bronchoalveolar Lavage Fluid
Bronchoalveolar Lavage
Indirect Fluorescent Antibody Technique
Flow Cytometry
Education

ASJC Scopus subject areas

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry
  • Cell Biology

Cite this

CD44 and CD69 represent different types of cell-surface activation markers for human eosinophils. / Matsumoto, Kenji; Appiah-Pippim, James Akwa; Schleimer, Robert P.; Bickel, Carol A.; Beck, Lisa A.; Bochner, Bruce S.

In: American journal of respiratory cell and molecular biology, Vol. 18, No. 6, 01.01.1998, p. 860-866.

Research output: Contribution to journalArticle

Matsumoto, Kenji ; Appiah-Pippim, James Akwa ; Schleimer, Robert P. ; Bickel, Carol A. ; Beck, Lisa A. ; Bochner, Bruce S. / CD44 and CD69 represent different types of cell-surface activation markers for human eosinophils. In: American journal of respiratory cell and molecular biology. 1998 ; Vol. 18, No. 6. pp. 860-866.
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abstract = "Eosinophils (EOS) purified from peripheral blood or late-phase bronchoalveolar lavage (BAL) were analyzed with 473 monoclonal antibodies (mAbs) from the Fifth International Workshop on Human Leukocyte Antigens in an attempt to identify markers of EOS activation. Two strategies were used: (1) to look for surface markers absent on fresh EOS but present after in vivo activation (e.g., in late-phase BAL fluid [BALF]) or after in vitro culture for up to 72 h with cytokines (≤ 10 ng/ml of interleukin-3 [IL-3], IL-5, or granulocyte-macrophage colony-stimulating factor [GM-CSF]); and (2) to look for markers constitutively expressed on fresh EOS that were increased after activation in vivo or after culture in vitro. With indirect immunofluorescence and flow cytometry, the first approach revealed that among approximately 350 mAbs tested, only those recognizing CD69 became bound to late-phase BALF EOS or cytokine-cultured EOS, but not to fresh EOS. Using the second approach, we observed statistically significant concentration-and time-dependent increases in CD44 expression in EOS cultured with IL-3, IL-5, or GM-CSF (∼ 2-fold increase in fluorescence intensity, P < 0.05), but not with interferon-γ (IFN-γ) (up to 100 ng/ml), whereas levels of 15 other constitutively expressed markers were unchanged. Despite increased expression, neither fresh nor cytokine-cultured EOS adhered to immobilized hyaluronate, a ligand for CD44. Additionally, simultaneous comparison of hypodense (specific gravity < 1.085 g/liter) and normodense (specific gravity > 1.085 g/liter) EOS from allergic donors consistently revealed higher levels of CD44 expression (∼ 3-to 8-fold) but not CD69 expression on hypodense EOS. We conclude that CD69 and CD44 represent different types of activation markers for human EOS. These findings may be useful in assessing the state of EOS activation in vitro and in vivo.",
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