Lack of detection of agonist activity by antibodies to platelet-derived growth factor receptor α in a subset of normal and systemic sclerosis patient Sera

Nick Loizos, Leah LaRiccia, Jami Weiner, Heather Griffith, Francesco Boin, Laura Hummers, Fredrick Wigley, Paul Kussie

Research output: Contribution to journalArticle

Abstract

To investigate whether agonist antiplatelet-derived growth factor receptor α (anti- PDGFRa) antibodies are present in the serum of patients with systemic sclerosis (SSc; scleroderma). Methods. Sera were obtained from healthy subjects and scleroderma patients. An electrochemiluminescence binding assay was performed for detection of serum autoantibodies to PDGFRa, PDGFRβ, epidermal growth factor receptor (EGFR), and colonystimulating factor receptor 1 (CSFR1). Serum immunoglobulin was purified by protein A/G chromatography. To assess Ig agonist activity, PDGFRa-expressing cells were incubated with pure Ig and the level of receptor phosphorylation determined in an enzyme-linked immunoassay, as well as by Western blotting. Ig agonist activity was also assessed in a mitogenic assay and by MAP kinase activation in a PDGFRa-expressing cell line. Results. Sera from 34.3% of the healthy subjects and 32.7% of the SSc patients contained detectable autoantibodies to PDGFRα and PDGFRβ, but not EGFR or CSFR1. Purified Ig from these sera was shown to retain PDGFR binding activity and, at 200-1,000 μg/ml, exhibited no agonist activity in a cell-based PDGFRa phosphorylation assay and did not stimulate a mitogenic response or MAP kinase activation in a PDGFRa-expressing cell line. Two purified Ig samples that were unable to bind PDGFRa did exhibit binding activity to a nonglycosylated form of PDGFRa. Conclusion. Although approximately one-third of sera from scleroderma patients contained detectable autoantibodies to PDGFR, these antibodies were not specific to scleroderma, since they were also detected in a similar percentage of samples from normal subjects. PDGFRa agonist activity was not demonstrated when purified Ig from these sera was tested in cell-based assays.

Original languageEnglish (US)
Pages (from-to)1145-1151
Number of pages7
JournalArthritis and Rheumatism
Volume60
Issue number4
DOIs
StatePublished - Apr 2009

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Platelet-Derived Growth Factor Receptors
Systemic Scleroderma
Antibodies
Serum
Autoantibodies
Epidermal Growth Factor Receptor
Healthy Volunteers
Phosphotransferases
Phosphorylation
Cell Line
Growth Factor Receptors
Staphylococcal Protein A
Immunoenzyme Techniques
Chromatography
Immunoglobulins
Anti-Idiotypic Antibodies
Western Blotting

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Rheumatology
  • Pharmacology (medical)

Cite this

Lack of detection of agonist activity by antibodies to platelet-derived growth factor receptor α in a subset of normal and systemic sclerosis patient Sera. / Loizos, Nick; LaRiccia, Leah; Weiner, Jami; Griffith, Heather; Boin, Francesco; Hummers, Laura; Wigley, Fredrick; Kussie, Paul.

In: Arthritis and Rheumatism, Vol. 60, No. 4, 04.2009, p. 1145-1151.

Research output: Contribution to journalArticle

Loizos, Nick ; LaRiccia, Leah ; Weiner, Jami ; Griffith, Heather ; Boin, Francesco ; Hummers, Laura ; Wigley, Fredrick ; Kussie, Paul. / Lack of detection of agonist activity by antibodies to platelet-derived growth factor receptor α in a subset of normal and systemic sclerosis patient Sera. In: Arthritis and Rheumatism. 2009 ; Vol. 60, No. 4. pp. 1145-1151.
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abstract = "To investigate whether agonist antiplatelet-derived growth factor receptor α (anti- PDGFRa) antibodies are present in the serum of patients with systemic sclerosis (SSc; scleroderma). Methods. Sera were obtained from healthy subjects and scleroderma patients. An electrochemiluminescence binding assay was performed for detection of serum autoantibodies to PDGFRa, PDGFRβ, epidermal growth factor receptor (EGFR), and colonystimulating factor receptor 1 (CSFR1). Serum immunoglobulin was purified by protein A/G chromatography. To assess Ig agonist activity, PDGFRa-expressing cells were incubated with pure Ig and the level of receptor phosphorylation determined in an enzyme-linked immunoassay, as well as by Western blotting. Ig agonist activity was also assessed in a mitogenic assay and by MAP kinase activation in a PDGFRa-expressing cell line. Results. Sera from 34.3{\%} of the healthy subjects and 32.7{\%} of the SSc patients contained detectable autoantibodies to PDGFRα and PDGFRβ, but not EGFR or CSFR1. Purified Ig from these sera was shown to retain PDGFR binding activity and, at 200-1,000 μg/ml, exhibited no agonist activity in a cell-based PDGFRa phosphorylation assay and did not stimulate a mitogenic response or MAP kinase activation in a PDGFRa-expressing cell line. Two purified Ig samples that were unable to bind PDGFRa did exhibit binding activity to a nonglycosylated form of PDGFRa. Conclusion. Although approximately one-third of sera from scleroderma patients contained detectable autoantibodies to PDGFR, these antibodies were not specific to scleroderma, since they were also detected in a similar percentage of samples from normal subjects. PDGFRa agonist activity was not demonstrated when purified Ig from these sera was tested in cell-based assays.",
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AU - Weiner, Jami

AU - Griffith, Heather

AU - Boin, Francesco

AU - Hummers, Laura

AU - Wigley, Fredrick

AU - Kussie, Paul

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N2 - To investigate whether agonist antiplatelet-derived growth factor receptor α (anti- PDGFRa) antibodies are present in the serum of patients with systemic sclerosis (SSc; scleroderma). Methods. Sera were obtained from healthy subjects and scleroderma patients. An electrochemiluminescence binding assay was performed for detection of serum autoantibodies to PDGFRa, PDGFRβ, epidermal growth factor receptor (EGFR), and colonystimulating factor receptor 1 (CSFR1). Serum immunoglobulin was purified by protein A/G chromatography. To assess Ig agonist activity, PDGFRa-expressing cells were incubated with pure Ig and the level of receptor phosphorylation determined in an enzyme-linked immunoassay, as well as by Western blotting. Ig agonist activity was also assessed in a mitogenic assay and by MAP kinase activation in a PDGFRa-expressing cell line. Results. Sera from 34.3% of the healthy subjects and 32.7% of the SSc patients contained detectable autoantibodies to PDGFRα and PDGFRβ, but not EGFR or CSFR1. Purified Ig from these sera was shown to retain PDGFR binding activity and, at 200-1,000 μg/ml, exhibited no agonist activity in a cell-based PDGFRa phosphorylation assay and did not stimulate a mitogenic response or MAP kinase activation in a PDGFRa-expressing cell line. Two purified Ig samples that were unable to bind PDGFRa did exhibit binding activity to a nonglycosylated form of PDGFRa. Conclusion. Although approximately one-third of sera from scleroderma patients contained detectable autoantibodies to PDGFR, these antibodies were not specific to scleroderma, since they were also detected in a similar percentage of samples from normal subjects. PDGFRa agonist activity was not demonstrated when purified Ig from these sera was tested in cell-based assays.

AB - To investigate whether agonist antiplatelet-derived growth factor receptor α (anti- PDGFRa) antibodies are present in the serum of patients with systemic sclerosis (SSc; scleroderma). Methods. Sera were obtained from healthy subjects and scleroderma patients. An electrochemiluminescence binding assay was performed for detection of serum autoantibodies to PDGFRa, PDGFRβ, epidermal growth factor receptor (EGFR), and colonystimulating factor receptor 1 (CSFR1). Serum immunoglobulin was purified by protein A/G chromatography. To assess Ig agonist activity, PDGFRa-expressing cells were incubated with pure Ig and the level of receptor phosphorylation determined in an enzyme-linked immunoassay, as well as by Western blotting. Ig agonist activity was also assessed in a mitogenic assay and by MAP kinase activation in a PDGFRa-expressing cell line. Results. Sera from 34.3% of the healthy subjects and 32.7% of the SSc patients contained detectable autoantibodies to PDGFRα and PDGFRβ, but not EGFR or CSFR1. Purified Ig from these sera was shown to retain PDGFR binding activity and, at 200-1,000 μg/ml, exhibited no agonist activity in a cell-based PDGFRa phosphorylation assay and did not stimulate a mitogenic response or MAP kinase activation in a PDGFRa-expressing cell line. Two purified Ig samples that were unable to bind PDGFRa did exhibit binding activity to a nonglycosylated form of PDGFRa. Conclusion. Although approximately one-third of sera from scleroderma patients contained detectable autoantibodies to PDGFR, these antibodies were not specific to scleroderma, since they were also detected in a similar percentage of samples from normal subjects. PDGFRa agonist activity was not demonstrated when purified Ig from these sera was tested in cell-based assays.

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