Preferential recognition of a fragment species of osteoarthritic synovial fluid fibronectin by antibodies to the alternatively spliced EIIIA segment

John H. Peters, Steven Carsons, Kenneth Kalunian, Skye McDougall, Mika Yoshida, Fred Ko, Milena Van Der Vliet-Hristova, Theodore J. Hahn

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Objective. To characterize the species of synovial fluid (SF) fibronectin (FN) bearing the alternatively spliced EIIIA segment. Methods. SF from patients with osteoarthritis (OA) and rheumatoid arthritis (RA), as well as corresponding affinity isolation products, were subjected to 1-dimensional and 2-dimensional electrophoresis followed by Western blot analysis. Results. Regardless of the clinical type of arthritis, a polyclonal antibody that recognizes antigenic determinants throughout the FN molecule produced staining of predominantly ∼ 200+ and ∼ 170-kd species in reduced 1-dimensional electrophoresis. Despite the overall prevalence of the larger species, 4 monoclonal antibodies (mAb) reactive with sequences lying near the center of the EIIIA segment exhibited a relative failure to recognize the larger of these 2 species in OA, but not RA, SF. The absence of recognition of EIIIA sequences within the ∼200+ kd forms of OA SF FN was unrelated to their derivation from dimers, since anti-EIIIA mAb recognized the smaller fragment species in preference to both monomeric and dimeric forms. The ∼170-kd EIIIA+ fragments were observed to have minimal gelatin-binding capacity and appeared on 2-dimensional electrophoresis to extend from the N-terminus of FN through at least the center of the EIIIA segment. Similar results were obtained for samples obtained by needle aspiration or arthroscopic lavage, suggesting a widespread applicability of these findings. Conclusion. The ∼170-kd EIIIA+ species of FN could potentially constitute a soluble "vehicle" by which chondrocyte-regulating EIIIA sequences, liberated from inhibitory flanking C-terminal sequences, could reach cells in the arthritic joint. Additionally, "FN species-specific" recognition of this segment within OA SF could constitute a marker by which to gauge the activity of the OA disease process.

Original languageEnglish (US)
Pages (from-to)2572-2585
Number of pages14
JournalArthritis and Rheumatism
Volume44
Issue number11
DOIs
StatePublished - 2001

Fingerprint

Synovial Fluid
Fibronectins
Osteoarthritis
Antibodies
Electrophoresis
Arthritis
Rheumatoid Arthritis
Monoclonal Antibodies
Therapeutic Irrigation
Gelatin
Chondrocytes
Needles
Epitopes
Joints
Western Blotting
Staining and Labeling

ASJC Scopus subject areas

  • Immunology
  • Rheumatology

Cite this

Preferential recognition of a fragment species of osteoarthritic synovial fluid fibronectin by antibodies to the alternatively spliced EIIIA segment. / Peters, John H.; Carsons, Steven; Kalunian, Kenneth; McDougall, Skye; Yoshida, Mika; Ko, Fred; Van Der Vliet-Hristova, Milena; Hahn, Theodore J.

In: Arthritis and Rheumatism, Vol. 44, No. 11, 2001, p. 2572-2585.

Research output: Contribution to journalArticle

Peters, John H. ; Carsons, Steven ; Kalunian, Kenneth ; McDougall, Skye ; Yoshida, Mika ; Ko, Fred ; Van Der Vliet-Hristova, Milena ; Hahn, Theodore J. / Preferential recognition of a fragment species of osteoarthritic synovial fluid fibronectin by antibodies to the alternatively spliced EIIIA segment. In: Arthritis and Rheumatism. 2001 ; Vol. 44, No. 11. pp. 2572-2585.
@article{c6f1652730d24b22922eca044da588a7,
title = "Preferential recognition of a fragment species of osteoarthritic synovial fluid fibronectin by antibodies to the alternatively spliced EIIIA segment",
abstract = "Objective. To characterize the species of synovial fluid (SF) fibronectin (FN) bearing the alternatively spliced EIIIA segment. Methods. SF from patients with osteoarthritis (OA) and rheumatoid arthritis (RA), as well as corresponding affinity isolation products, were subjected to 1-dimensional and 2-dimensional electrophoresis followed by Western blot analysis. Results. Regardless of the clinical type of arthritis, a polyclonal antibody that recognizes antigenic determinants throughout the FN molecule produced staining of predominantly ∼ 200+ and ∼ 170-kd species in reduced 1-dimensional electrophoresis. Despite the overall prevalence of the larger species, 4 monoclonal antibodies (mAb) reactive with sequences lying near the center of the EIIIA segment exhibited a relative failure to recognize the larger of these 2 species in OA, but not RA, SF. The absence of recognition of EIIIA sequences within the ∼200+ kd forms of OA SF FN was unrelated to their derivation from dimers, since anti-EIIIA mAb recognized the smaller fragment species in preference to both monomeric and dimeric forms. The ∼170-kd EIIIA+ fragments were observed to have minimal gelatin-binding capacity and appeared on 2-dimensional electrophoresis to extend from the N-terminus of FN through at least the center of the EIIIA segment. Similar results were obtained for samples obtained by needle aspiration or arthroscopic lavage, suggesting a widespread applicability of these findings. Conclusion. The ∼170-kd EIIIA+ species of FN could potentially constitute a soluble {"}vehicle{"} by which chondrocyte-regulating EIIIA sequences, liberated from inhibitory flanking C-terminal sequences, could reach cells in the arthritic joint. Additionally, {"}FN species-specific{"} recognition of this segment within OA SF could constitute a marker by which to gauge the activity of the OA disease process.",
author = "Peters, {John H.} and Steven Carsons and Kenneth Kalunian and Skye McDougall and Mika Yoshida and Fred Ko and {Van Der Vliet-Hristova}, Milena and Hahn, {Theodore J.}",
year = "2001",
doi = "10.1002/1529-0131(200111)44:11<2572::AID-ART438>3.0.CO;2-Y",
language = "English (US)",
volume = "44",
pages = "2572--2585",
journal = "Arthritis and Rheumatology",
issn = "2326-5191",
publisher = "John Wiley and Sons Ltd",
number = "11",

}

TY - JOUR

T1 - Preferential recognition of a fragment species of osteoarthritic synovial fluid fibronectin by antibodies to the alternatively spliced EIIIA segment

AU - Peters, John H.

AU - Carsons, Steven

AU - Kalunian, Kenneth

AU - McDougall, Skye

AU - Yoshida, Mika

AU - Ko, Fred

AU - Van Der Vliet-Hristova, Milena

AU - Hahn, Theodore J.

PY - 2001

Y1 - 2001

N2 - Objective. To characterize the species of synovial fluid (SF) fibronectin (FN) bearing the alternatively spliced EIIIA segment. Methods. SF from patients with osteoarthritis (OA) and rheumatoid arthritis (RA), as well as corresponding affinity isolation products, were subjected to 1-dimensional and 2-dimensional electrophoresis followed by Western blot analysis. Results. Regardless of the clinical type of arthritis, a polyclonal antibody that recognizes antigenic determinants throughout the FN molecule produced staining of predominantly ∼ 200+ and ∼ 170-kd species in reduced 1-dimensional electrophoresis. Despite the overall prevalence of the larger species, 4 monoclonal antibodies (mAb) reactive with sequences lying near the center of the EIIIA segment exhibited a relative failure to recognize the larger of these 2 species in OA, but not RA, SF. The absence of recognition of EIIIA sequences within the ∼200+ kd forms of OA SF FN was unrelated to their derivation from dimers, since anti-EIIIA mAb recognized the smaller fragment species in preference to both monomeric and dimeric forms. The ∼170-kd EIIIA+ fragments were observed to have minimal gelatin-binding capacity and appeared on 2-dimensional electrophoresis to extend from the N-terminus of FN through at least the center of the EIIIA segment. Similar results were obtained for samples obtained by needle aspiration or arthroscopic lavage, suggesting a widespread applicability of these findings. Conclusion. The ∼170-kd EIIIA+ species of FN could potentially constitute a soluble "vehicle" by which chondrocyte-regulating EIIIA sequences, liberated from inhibitory flanking C-terminal sequences, could reach cells in the arthritic joint. Additionally, "FN species-specific" recognition of this segment within OA SF could constitute a marker by which to gauge the activity of the OA disease process.

AB - Objective. To characterize the species of synovial fluid (SF) fibronectin (FN) bearing the alternatively spliced EIIIA segment. Methods. SF from patients with osteoarthritis (OA) and rheumatoid arthritis (RA), as well as corresponding affinity isolation products, were subjected to 1-dimensional and 2-dimensional electrophoresis followed by Western blot analysis. Results. Regardless of the clinical type of arthritis, a polyclonal antibody that recognizes antigenic determinants throughout the FN molecule produced staining of predominantly ∼ 200+ and ∼ 170-kd species in reduced 1-dimensional electrophoresis. Despite the overall prevalence of the larger species, 4 monoclonal antibodies (mAb) reactive with sequences lying near the center of the EIIIA segment exhibited a relative failure to recognize the larger of these 2 species in OA, but not RA, SF. The absence of recognition of EIIIA sequences within the ∼200+ kd forms of OA SF FN was unrelated to their derivation from dimers, since anti-EIIIA mAb recognized the smaller fragment species in preference to both monomeric and dimeric forms. The ∼170-kd EIIIA+ fragments were observed to have minimal gelatin-binding capacity and appeared on 2-dimensional electrophoresis to extend from the N-terminus of FN through at least the center of the EIIIA segment. Similar results were obtained for samples obtained by needle aspiration or arthroscopic lavage, suggesting a widespread applicability of these findings. Conclusion. The ∼170-kd EIIIA+ species of FN could potentially constitute a soluble "vehicle" by which chondrocyte-regulating EIIIA sequences, liberated from inhibitory flanking C-terminal sequences, could reach cells in the arthritic joint. Additionally, "FN species-specific" recognition of this segment within OA SF could constitute a marker by which to gauge the activity of the OA disease process.

UR - http://www.scopus.com/inward/record.url?scp=0035149058&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035149058&partnerID=8YFLogxK

U2 - 10.1002/1529-0131(200111)44:11<2572::AID-ART438>3.0.CO;2-Y

DO - 10.1002/1529-0131(200111)44:11<2572::AID-ART438>3.0.CO;2-Y

M3 - Article

C2 - 11710714

AN - SCOPUS:0035149058

VL - 44

SP - 2572

EP - 2585

JO - Arthritis and Rheumatology

JF - Arthritis and Rheumatology

SN - 2326-5191

IS - 11

ER -