HUMAN EOSINOPHILS PREFERENTIALLY SURVIVE ON TISSUE FIBRONECTIN COMPARED WITH PLASMA FIBRONECTIN

Garry Michael Walsh, F A SYMON, A J WARDLAW

Research output: Contribution to journalArticle

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Abstract

Background Eosinophil-derived inflammatory mediators including cytokines are considered to be important in the pathogenesis of allergic inflammation. Fibronectin (Fn) has been shown to be a physiological trigger of autocrine cytokine production by human eosinophils. Fn is encoded by a single gene, but alternate splicing of the primary RNA transcript results in polypeptide diversity in a cell type-specific fashion. Thus, tissue Fn contains approximately 50% more of the CS-1 cell binding region recognized by the integrin alpha 4 beta 1 compared with plasma Fn.

Objective Since eosinophils are predominantly tissue-dwelling cells we compared the effect of tissue and plasma Fn on eosinophil survival in culture.

Methods The viability and cytokine generation of eosinophils (>99.9% pure) cultured for up to 4 days in 96 well plates coated with tissue Fn, plasma Fn or BSA was compared.

Results There was a significant difference in the ability of tissue Fn to support eosinophil survival compared with plasma Fn (P < 0.01). Optimal survival with tissue Fn was seen at 25 mu g/well (70% +/- 2.0% viability at 3 days vs 7% +/- 2.2% viability on BSA). Significant (P < 0.001) cell viability on tissue Fn was observed for up to 4 days in culture (54% +/- 6.0%) compared with BSA coated wells. Addition of autologous mononuclear cells (final concentration 0.5%, 1% or 2%) resulted in plasma Fn-dependent eosinophil survival comparable to that of 99.9% pure eosinophils adherent to tissue Fn. Tissue Fn-dependent survival was significantly inhibited by anti-interleukin-3, anti-granulocyte macrophage colony stimulating factor (GM-CSF) and anti-IL-5 monoclonal antibodies. Picogram quantities of these three cytokines were detected in supernatants from eosinophils cultured for 3 days on tissue Fn using specific enzyme-linked immunosorbent assays (ELISAs). Eosinophil survival on tissue Fn was significantly inhibited by anti-pr and alpha 4 beta 1 monoclonal antibody (MoAb) and also by a MoAb specific for the CS-1 motif in the IIICS region of Fn.

Conclusion These observations show preferential survival of eosinophils cultured on tissue Fn as a result of alpha 4 beta 1-dependent interaction with the CS-1 region of tissue Fn triggering autocrine cytokine synthesis and release, thereby promoting their survival and persistence within the tissues.

Original languageEnglish
Pages (from-to)1128-1136
Number of pages9
JournalClinical & experimental allergy
Volume25
Issue number11
Publication statusPublished - Nov 1995

Keywords

  • EOSINOPHIL
  • CELL SURVIVAL
  • ASTHMA
  • FIBRONECTIN
  • EXTRACELLULAR MATRIX
  • CYTOKINES
  • COLONY-STIMULATING FACTOR
  • MESSENGER-RNA
  • BRONCHIAL BIOPSIES
  • BLOOD EOSINOPHILS
  • ADHESION RECEPTOR
  • 3T3 FIBROBLASTS
  • LYMPHOCYTES-T
  • IIICS REGION
  • IDENTIFICATION

Cite this

HUMAN EOSINOPHILS PREFERENTIALLY SURVIVE ON TISSUE FIBRONECTIN COMPARED WITH PLASMA FIBRONECTIN. / Walsh, Garry Michael; SYMON, F A ; WARDLAW, A J .

In: Clinical & experimental allergy, Vol. 25, No. 11, 11.1995, p. 1128-1136.

Research output: Contribution to journalArticle

@article{76a4adcc8e1a461ca599ec8aa29650ca,
title = "HUMAN EOSINOPHILS PREFERENTIALLY SURVIVE ON TISSUE FIBRONECTIN COMPARED WITH PLASMA FIBRONECTIN",
abstract = "Background Eosinophil-derived inflammatory mediators including cytokines are considered to be important in the pathogenesis of allergic inflammation. Fibronectin (Fn) has been shown to be a physiological trigger of autocrine cytokine production by human eosinophils. Fn is encoded by a single gene, but alternate splicing of the primary RNA transcript results in polypeptide diversity in a cell type-specific fashion. Thus, tissue Fn contains approximately 50{\%} more of the CS-1 cell binding region recognized by the integrin alpha 4 beta 1 compared with plasma Fn.Objective Since eosinophils are predominantly tissue-dwelling cells we compared the effect of tissue and plasma Fn on eosinophil survival in culture.Methods The viability and cytokine generation of eosinophils (>99.9{\%} pure) cultured for up to 4 days in 96 well plates coated with tissue Fn, plasma Fn or BSA was compared.Results There was a significant difference in the ability of tissue Fn to support eosinophil survival compared with plasma Fn (P < 0.01). Optimal survival with tissue Fn was seen at 25 mu g/well (70{\%} +/- 2.0{\%} viability at 3 days vs 7{\%} +/- 2.2{\%} viability on BSA). Significant (P < 0.001) cell viability on tissue Fn was observed for up to 4 days in culture (54{\%} +/- 6.0{\%}) compared with BSA coated wells. Addition of autologous mononuclear cells (final concentration 0.5{\%}, 1{\%} or 2{\%}) resulted in plasma Fn-dependent eosinophil survival comparable to that of 99.9{\%} pure eosinophils adherent to tissue Fn. Tissue Fn-dependent survival was significantly inhibited by anti-interleukin-3, anti-granulocyte macrophage colony stimulating factor (GM-CSF) and anti-IL-5 monoclonal antibodies. Picogram quantities of these three cytokines were detected in supernatants from eosinophils cultured for 3 days on tissue Fn using specific enzyme-linked immunosorbent assays (ELISAs). Eosinophil survival on tissue Fn was significantly inhibited by anti-pr and alpha 4 beta 1 monoclonal antibody (MoAb) and also by a MoAb specific for the CS-1 motif in the IIICS region of Fn.Conclusion These observations show preferential survival of eosinophils cultured on tissue Fn as a result of alpha 4 beta 1-dependent interaction with the CS-1 region of tissue Fn triggering autocrine cytokine synthesis and release, thereby promoting their survival and persistence within the tissues.",
keywords = "EOSINOPHIL, CELL SURVIVAL, ASTHMA, FIBRONECTIN, EXTRACELLULAR MATRIX, CYTOKINES, COLONY-STIMULATING FACTOR, MESSENGER-RNA, BRONCHIAL BIOPSIES, BLOOD EOSINOPHILS, ADHESION RECEPTOR, 3T3 FIBROBLASTS, LYMPHOCYTES-T, IIICS REGION, IDENTIFICATION",
author = "Walsh, {Garry Michael} and SYMON, {F A} and WARDLAW, {A J}",
year = "1995",
month = "11",
language = "English",
volume = "25",
pages = "1128--1136",
journal = "Clinical & experimental allergy",
issn = "0954-7894",
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T1 - HUMAN EOSINOPHILS PREFERENTIALLY SURVIVE ON TISSUE FIBRONECTIN COMPARED WITH PLASMA FIBRONECTIN

AU - Walsh, Garry Michael

AU - SYMON, F A

AU - WARDLAW, A J

PY - 1995/11

Y1 - 1995/11

N2 - Background Eosinophil-derived inflammatory mediators including cytokines are considered to be important in the pathogenesis of allergic inflammation. Fibronectin (Fn) has been shown to be a physiological trigger of autocrine cytokine production by human eosinophils. Fn is encoded by a single gene, but alternate splicing of the primary RNA transcript results in polypeptide diversity in a cell type-specific fashion. Thus, tissue Fn contains approximately 50% more of the CS-1 cell binding region recognized by the integrin alpha 4 beta 1 compared with plasma Fn.Objective Since eosinophils are predominantly tissue-dwelling cells we compared the effect of tissue and plasma Fn on eosinophil survival in culture.Methods The viability and cytokine generation of eosinophils (>99.9% pure) cultured for up to 4 days in 96 well plates coated with tissue Fn, plasma Fn or BSA was compared.Results There was a significant difference in the ability of tissue Fn to support eosinophil survival compared with plasma Fn (P < 0.01). Optimal survival with tissue Fn was seen at 25 mu g/well (70% +/- 2.0% viability at 3 days vs 7% +/- 2.2% viability on BSA). Significant (P < 0.001) cell viability on tissue Fn was observed for up to 4 days in culture (54% +/- 6.0%) compared with BSA coated wells. Addition of autologous mononuclear cells (final concentration 0.5%, 1% or 2%) resulted in plasma Fn-dependent eosinophil survival comparable to that of 99.9% pure eosinophils adherent to tissue Fn. Tissue Fn-dependent survival was significantly inhibited by anti-interleukin-3, anti-granulocyte macrophage colony stimulating factor (GM-CSF) and anti-IL-5 monoclonal antibodies. Picogram quantities of these three cytokines were detected in supernatants from eosinophils cultured for 3 days on tissue Fn using specific enzyme-linked immunosorbent assays (ELISAs). Eosinophil survival on tissue Fn was significantly inhibited by anti-pr and alpha 4 beta 1 monoclonal antibody (MoAb) and also by a MoAb specific for the CS-1 motif in the IIICS region of Fn.Conclusion These observations show preferential survival of eosinophils cultured on tissue Fn as a result of alpha 4 beta 1-dependent interaction with the CS-1 region of tissue Fn triggering autocrine cytokine synthesis and release, thereby promoting their survival and persistence within the tissues.

AB - Background Eosinophil-derived inflammatory mediators including cytokines are considered to be important in the pathogenesis of allergic inflammation. Fibronectin (Fn) has been shown to be a physiological trigger of autocrine cytokine production by human eosinophils. Fn is encoded by a single gene, but alternate splicing of the primary RNA transcript results in polypeptide diversity in a cell type-specific fashion. Thus, tissue Fn contains approximately 50% more of the CS-1 cell binding region recognized by the integrin alpha 4 beta 1 compared with plasma Fn.Objective Since eosinophils are predominantly tissue-dwelling cells we compared the effect of tissue and plasma Fn on eosinophil survival in culture.Methods The viability and cytokine generation of eosinophils (>99.9% pure) cultured for up to 4 days in 96 well plates coated with tissue Fn, plasma Fn or BSA was compared.Results There was a significant difference in the ability of tissue Fn to support eosinophil survival compared with plasma Fn (P < 0.01). Optimal survival with tissue Fn was seen at 25 mu g/well (70% +/- 2.0% viability at 3 days vs 7% +/- 2.2% viability on BSA). Significant (P < 0.001) cell viability on tissue Fn was observed for up to 4 days in culture (54% +/- 6.0%) compared with BSA coated wells. Addition of autologous mononuclear cells (final concentration 0.5%, 1% or 2%) resulted in plasma Fn-dependent eosinophil survival comparable to that of 99.9% pure eosinophils adherent to tissue Fn. Tissue Fn-dependent survival was significantly inhibited by anti-interleukin-3, anti-granulocyte macrophage colony stimulating factor (GM-CSF) and anti-IL-5 monoclonal antibodies. Picogram quantities of these three cytokines were detected in supernatants from eosinophils cultured for 3 days on tissue Fn using specific enzyme-linked immunosorbent assays (ELISAs). Eosinophil survival on tissue Fn was significantly inhibited by anti-pr and alpha 4 beta 1 monoclonal antibody (MoAb) and also by a MoAb specific for the CS-1 motif in the IIICS region of Fn.Conclusion These observations show preferential survival of eosinophils cultured on tissue Fn as a result of alpha 4 beta 1-dependent interaction with the CS-1 region of tissue Fn triggering autocrine cytokine synthesis and release, thereby promoting their survival and persistence within the tissues.

KW - EOSINOPHIL

KW - CELL SURVIVAL

KW - ASTHMA

KW - FIBRONECTIN

KW - EXTRACELLULAR MATRIX

KW - CYTOKINES

KW - COLONY-STIMULATING FACTOR

KW - MESSENGER-RNA

KW - BRONCHIAL BIOPSIES

KW - BLOOD EOSINOPHILS

KW - ADHESION RECEPTOR

KW - 3T3 FIBROBLASTS

KW - LYMPHOCYTES-T

KW - IIICS REGION

KW - IDENTIFICATION

M3 - Article

VL - 25

SP - 1128

EP - 1136

JO - Clinical & experimental allergy

JF - Clinical & experimental allergy

SN - 0954-7894

IS - 11

ER -