Characterization of Human Recombinant Interleukin 2 Binding to Heparin and Heparan Sulfate Using an Elisa Approach

Najjam, Saloua; Gibbs, Roslyn V.; Gordon, Myrtle Y.; Rider, Christopher C.

doi:10.1006/cyto.1997.0246

Cited by 90 publications

(69 citation statements)

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“…The more LMWH the sample contains, the longer the interaction lasts, the broader the IL-2 peak becomes and the lower the peak height changes. The peak heights of IL-2 were used to estimate the binding constant of the interaction between IL-2 and LMWH, and the value (1.2 × 10 6 M −1 ) obtained here is in good agreement with that obtained by ELISA (2.0 × 10 6 M −1 ) [29]. This demonstrated that the fast on-and-off kinetic interaction systems could also be studied by CZE contrary to traditional opinion that CZE could only be used in studying slow onand-off kinetic interactions and ACE is exclusive in studying fast on-and-off ones based on CE methods.…”

Section: Discussionsupporting

confidence: 81%

“…(1). From the slope of the Scatchard plot, K was calculated to be 1.2 × 10 6 M −1 which was in excellent agreement with the K D value of 0.5 M (that means K is 2.0 × 10 6 M −1 ) that Saloua et al obtained by using ELISA [29]. This demonstrated that the fast on-and-off rates interactions might be studied using CZE.…”

Section: The Interaction Between Il-2 and Lmwhsupporting

confidence: 79%

“…The interaction between IL-2 and heparin has been investigated by an enzyme-linked immunosorbent assay (ELISA) [1,29]. IL-2 binds to heparin in a dose-dependent manner.…”

Section: Introductionmentioning

confidence: 99%

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Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2

Liang

et al. 2005

Journal of Pharmaceutical and Biomedical Analysis

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Section: Discussionsupporting

confidence: 81%

Section: The Interaction Between Il-2 and Lmwhsupporting

confidence: 79%

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Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2

Liang

et al. 2005

Journal of Pharmaceutical and Biomedical Analysis

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“…Binding of ␤2-GPI to immobilized heparin was performed using a modification of the method of Najjam and co-workers (28). Nunc Maxisorp plates were incubated overnight at 4°C with heparin-BSA complex or BSA (Sigma) alone at a concentration of 5.0 g/ml in 13 mM EDTA and 50 mM Tris-HCl, pH 7.4.…”

Section: Methodsmentioning

confidence: 99%

Heparin Inhibits the Binding of β2-glycoprotein I to Phospholipids and Promotes the Plasmin-mediated Inactivation of This Blood Protein

Guerin

Sheng

Reddel

et al. 2002

Journal of Biological Chemistry

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The phospholipid-binding plasma protein ␤2-glycoprotein I (␤2-GPI) is the primary antigen recognized by the circulating autoantibodies in patients with the "anti-phospholipid syndrome" (APS). Although heparin is routinely used in the treatment and prophylaxis of APS patients, the primary heparin-binding site within ␤2-GPI has not been identified. More importantly, how heparin exerts its beneficial effects in vivo in APS patients has not been deduced at the molecular level. Using an expression/site-directed mutagenesis approach, we now show that the positively charged site that resides in the first domain of ␤2-GPI is not the primary heparin-binding site. Rather it is the second positively charged site located within the fifth domain of the protein that also binds to phospholipids. Lys 284 , Lys 286 , and Lys 287 in this domain are essential for the interaction of ␤2-GPI with heparin. These data indicate that ␤2-GPI binds to heparin in a relatively specific manner even though the affinity for the interaction is rather low. Lys 317 resides in the center of the high affinity phospholipid-binding site. Surprisingly, heparin at concentrations that can be achieved in vivo during anticoagulation therapy greatly enhances the plasmin-mediated cleavage of the Lys 317 -Thr 318 site in ␤2-GPI. Because the cleaved form cannot bind to phospholipids effectively, the combined actions of heparin and plasmin result in a diminished ability of ␤2-GPI to recognize phospholipids. This, in turn, decreases the prothrombotic activity of the endogenous circulating anti-␤2-GPI antibodies in the patients. Thus, heparin exerts its beneficial effects in APS patients by at least two distinct mechanisms.

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“…However, little research has been conducted into the possible biological significance of sulphated polysaccharides found in green algae. The present study was undertaken to evaluate the nature and possible therapeutic potential of the interactions between a sulphated polysaccharide structure extracted from the green alga Codium fragile and the three immune cytokines interleukin-2 (IL-2), interleukin-7 (IL-7) and gamma interferon (INF-y), all of which have been characterized as heparin-binding cytokines (Clarke et al, 1995;Lortat-Jacob et al, 1996;Najjam et al, 1997) and evaluated as therapeutic agents. The ability of other commercial polysaccharides extracted from brown (fucoidan) and red (carrageenan) algae to bind to the same cytokines was also assessed.…”

Section: Introductionmentioning

confidence: 99%

Specific recognition of immune cytokines by sulphated polysaccharides from marine algae

Nika

Carpenter

Gibbs

2003

European Journal of Phycology

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Several immune cytokines, including interleukin-2 (IL-2), interleukin-7 (IL-7) and gamma interferon (INF-y), have been shown to interact with the sulphated polysaccharides heparin and heparan sulphate, so that the co-administration of cytokines with such sulphated sugars may improve current cytokine-based therapies. In this study, sulphated polysaccharides were extracted from the green alga Codiumfragile in cold water and purified by size exclusion chromatography. Gas chromatography~massspectroscopy indicated that they were galactans and/or arabinogalactans, and preliminary nuclear magnetic resonance spectroscopy analysis suggested that they consisted of repeating units of several monosaccharide residues, probably galactopyranose and arabinopyranose, which were sulphated. The binding of Codium fragile polysaccharIde and commercial preparations of fucoidan, carrageenans and heparin to IL-2. IL-7 and INF-}' was then assessed using an ELISA-based inhibition test. For all cytokines tested, inhibition by fucoidan was found to be strongest (80-90% inhibition), followed by heparin (65-80% inhibition), the Codium polysaccharide (40-59% inhibition) and finally the carrageenans (20-60% inhibition). The differential binding profiles obtained for the cytokines tested suggest the interactions between cytokines and sulphated polysaccharides are specific and could therefore be exploited in a therapeutic capacity.

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Characterization of Human Recombinant Interleukin 2 Binding to Heparin and Heparan Sulfate Using an Elisa Approach

Cited by 90 publications

References 0 publications

Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2

Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2

Heparin Inhibits the Binding of β2-glycoprotein I to Phospholipids and Promotes the Plasmin-mediated Inactivation of This Blood Protein

Specific recognition of immune cytokines by sulphated polysaccharides from marine algae

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