EphB Receptor-binding Peptides Identified by Phage Display Enable Design of an Antagonist with Ephrin-like Affinity

Koolpe, Mitchell; Burgess, Rosemary; Dail, Monique; Pasquale, Elena B.

doi:10.1074/jbc.m500363200

Cited by 131 publications

(162 citation statements)

References 49 publications

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“…LA (55) was used at 2.5 g/ml for 30 min prior to the addition of cytokines; STAT3 inhibitor (ML116) (56) was used for 1 h prior to the addition of cytokines at 0.1, 1, or 10 g/ml. Cells were then washed twice with ice-cold PBS before cell lysates were prepared by shaking at 4°C with modified radioimmune precipitation assay buffer (84), proteinase inhibitor (Sigma), and phosphatase inhibitor (Dako) before lysates were centrifuged. Protein concentration was determined according to the manufacturer's protocol (Pierce).…”

Section: Methodsmentioning

confidence: 99%

Murine Oncostatin M Acts via Leukemia Inhibitory Factor Receptor to Phosphorylate Signal Transducer and Activator of Transcription 3 (STAT3) but Not STAT1, an Effect That Protects Bone Mass

Walker

Johnson

et al. 2016

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

Murine Oncostatin M Acts via Leukemia Inhibitory Factor Receptor to Phosphorylate Signal Transducer and Activator of Transcription 3 (STAT3) but Not STAT1, an Effect That Protects Bone Mass

Walker

Johnson

et al. 2016

Journal of Biological Chemistry

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“…The Eph family of receptor tyrosine kinases is an attractive candidate for therapeutic targets. Previous studies have developed a monoclonal antibody, peptide or small molecules against EphB2 in an attempt to block its activation (17)(18)(19), which may be applied as a potential remedy in pancreatic cancer as there are few treatment options available for patients with a disease with such a high mortality rate, resulting in poor outcomes.…”

Section: -B2 --------------------------------------------------------mentioning

confidence: 99%

Overexpression of the B-type Eph and ephrin genes correlates with progression and pain in human pancreatic cancer

Lu¹,

Zhang²,

Li³

et al. 2012

Oncology Letters

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Abstract. B-type erythropoietin-producing hepatocellular (EphB) receptors and their ephrin ligands are associated with aggressive behavior and poor prognosis in a number of malignancies. The aim of this study was to determine the association between the aberrant expression of these genes and prognosis in human pancreatic cancer. The expression of EphB and the ephrin ligands was determined using real-time quantitative reverse transcription PCR in 46 human primary pancreatic cancers. Overexpression of EphB2 and a more modest overexpression of ephrin-B2 mRNA were observed in more than 44% (20/46) of the pancreatic cancer specimens examined. Overexpression (>upper quartile) of EphB2 and ephrin-B2 was markedly associated with abdominal and/or back pain. Multivariate analysis identified the overexpression of EphB2 as an independent prognostic factor for disease-free and overall survival (both P<0.05). The results of the present study suggest that an increased level of ephrin-B2, in the presence of a high expression of EphB2, leads to a more aggressive tumor phenotype and that EphB2 may be used as a prognostic factor in human pancreatic cancer.

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“…The functional role of EphB2 and ephrin-B2 in inhibiting T-cell proliferation was further substantiated using blocking peptides, which have previously been shown to effectively block the ligandbinding site of specific Eph receptors [14,50]. In addition, these blocking peptides not only act to block stabilized activation of the EphB receptor, but can also inhibit simultaneous activation of ephrin-B reverse signaling [14,[49][50][51][52][53][54]. The present study demonstrated that T-cells exhibited increased proliferation in the presence of EphB2 or EphB4 blocking peptides compared with the scrambled peptide control.…”

Section: Fig 6 Msc Inhibitory Effect Of T-cell Proliferation Is Medmentioning

confidence: 99%

EphB and ephrin-B interactions mediate human mesenchymal stem cell suppression of activated T-cells

Nguyen

Gronthos

Arthur

et al. 2013

Experimental Hematology

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Mesenchymal stromal/stem cells (MSC) express the contact-dependent erythropoietin-producing hepatocellular (Eph) receptor tyrosine kinase family and their cognate ephrin ligands, which are known to regulate thymocyte maturation and selection, T-cell transendothelial migration, activation, co-stimulation, and proliferation. However, the contribution of Eph/ephrin molecules in mediating human MSC suppression of activated T-cells remains to be determined. In the present study, we showed that EphB2 and ephrin-B2 are expressed by ex vivo expanded MSC, while the corresponding ligands, ephrin-B1 and EphB4, respectively, are highly expressed by T-cells. Initial studies demonstrated that EphB2-Fc and ephrin-B2-Fc molecules suppressed T-cell proliferation in allogeneic mixed lymphocyte reaction (MLR) assays compared with human IgG-treated controls. While the addition of a third-party MSC population demonstrated dramatic suppression of T-cell proliferation responses in the MLR, blocking the function of EphB2 or EphB4 receptors using inhibitor binding peptides significantly increased T-cell proliferation. Consistent with these observations, shRNA EphB2 or ephrin-B2 knockdown expression in MSC reduced their ability to inhibit T-cell proliferation. Importantly, the expression of immunosuppressive factors, indoleamine 2, 3-dioxygenase, transforming growth factor-b1, and inducible nitric oxide synthase expressed by MSC, was up-regulated after stimulation with EphB4 and ephrin-B1 in the presence of interferon (IFN)-g, compared with untreated controls. Conversely, key factors involved in T-cell activation and proliferation, such as interleukin (IL)-2, IFN-g, tumor necrosis factor-a, and IL-17, were down-regulated by T-cells treated with EphB2 or ephrin-B2 compared with untreated controls. Studies utilizing signaling inhibitors revealed that inhibition of T-cell proliferation is partly mediated through EphB2-induced ephrin-B1 reverse signaling or ephrin-B2-mediated EphB4 forward signaling by activating Src, PI3Kinase, Abl, and JNK kinase pathways, activated by tyrosine phosphorylation. Taken together, these observations suggest that EphB/ephrin-B interactions play an important role in mediating human MSC inhibition of activated T cells.

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EphB Receptor-binding Peptides Identified by Phage Display Enable Design of an Antagonist with Ephrin-like Affinity

Cited by 131 publications

References 49 publications

Murine Oncostatin M Acts via Leukemia Inhibitory Factor Receptor to Phosphorylate Signal Transducer and Activator of Transcription 3 (STAT3) but Not STAT1, an Effect That Protects Bone Mass

Murine Oncostatin M Acts via Leukemia Inhibitory Factor Receptor to Phosphorylate Signal Transducer and Activator of Transcription 3 (STAT3) but Not STAT1, an Effect That Protects Bone Mass

Overexpression of the B-type Eph and ephrin genes correlates with progression and pain in human pancreatic cancer

EphB and ephrin-B interactions mediate human mesenchymal stem cell suppression of activated T-cells

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