recombinant osk1 peptide  (Alomone Labs)

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Potency of different Kv1.3 blockers on human Kv1.3 channels on transfected CHO cells.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Transfection

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: OsK1 peptide inhibited human Kv1.3 mediated whole cell currents in a concentration dependent manner in human CD4 T cells (A). Potency of OsK1 peptide on Kv1.3 currents in purified human CD4 T cells and Kv1.3 transfected CHO cells were determined and IC 50 curves were shown (B). Whole cell voltage-gated currents were measured in manual patch clamp assays. All recordings were made at room temperature using Multiclamp 700A amplifier and pClamp 9 software as described in Methods and Materials.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Concentration Assay, Purification, Transfection, Patch Clamp, Software

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Potency and efficacy of Kv1.3 blockers on human T cells under different stimulation conditions.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Inhibition

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Kv1.3 mRNA expression in different human tissues was profiled in Affymetrix microarrays (A). Cell surface Kv1.3 channels were measured with patch express assays on activated or resting human CD4 T cell subsets including naïve, central memory (Tcm) and effector memory (Tem) T cells, as well as antigen-specific T cells from house dust mite (HDM) stimulation. Data shown are either channels/cell (B) or number of channels normalized to cell surface area (channels/pF) (C). Time course of Kv1.3 channel activity and expression of the activation markers CD25 and CD69 are shown on effector memory CD4 T cells activated with anti-CD3/CD28 coated beads (D). Expression of Kv1.3 mRNA levels in naïve, Tcm and Tem CD4 T cells at different time points post activation with anti-CD3/CD28 coated beads are presented (E). Patch express measurement of Kv1.3 channels was performed on T cell subsets purified from 6 healthy blood donors. Statistical significance on the difference in Kv1.3 channel levels between activated and resting T cell subsets was analyzed by 2-tailed Student’s t test and P values were determined and indicated as *P < 0.05, ***P < 0.001, ****P < 0.0001.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Expressing, Activity Assay, Activation Assay, Purification

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Kv1.3 expression on human primary T cell subsets.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Expressing

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Kv1.3 blocker OsK1, Kv261 peptides and Kv261-HSA-34 fusion protein blocked proliferation of CD4 T cells from house dust mite allergic patients following 5 day stimulation with house dust mite extracts (A, B). IL-5 production from house dust mite-specific T cells was inhibited by Kv1.3 blockers as shown with OsK1 and Kv261-HSA-34 (C). OsK1 peptide at 0.3μM inhibited partially the proliferation of PBMC from patients allergic to ragweed after 3 day stimulation with ragweed extracts (D). Decreased proliferative response of individual donor PBMC samples by OsK1 peptide was shown in a separated graph. CTLA4-Ig or cyclosporine A (1μM) were used as reference compounds. The assays in panels A-C were repeated in 4 independent experiments with samples in duplicates. Data from one representative experiment were presented. The study in panel D was done in an experiment with 29 subjects per group. Statistical significance on the difference in proliferation between untreated and OsK1 treated groups were analyzed by 2-way ANOVA and the P value was determined and indicated as *P < 0.05. Readouts in (A) and (C) were performed on CD4 T cells from the same donor whereas those in (B) were from a separate donor. Data from one representative experiment were presented.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques:

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Potency of different Kv1.3 blockers on human Kv1.3 channels on transfected CHO cells.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Transfection

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: OsK1 peptide inhibited human Kv1.3 mediated whole cell currents in a concentration dependent manner in human CD4 T cells (A). Potency of OsK1 peptide on Kv1.3 currents in purified human CD4 T cells and Kv1.3 transfected CHO cells were determined and IC 50 curves were shown (B). Whole cell voltage-gated currents were measured in manual patch clamp assays. All recordings were made at room temperature using Multiclamp 700A amplifier and pClamp 9 software as described in Methods and Materials.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Concentration Assay, Purification, Transfection, Patch Clamp, Software

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Effect of OsK1 peptide (filled circles) or cyclosporine A (open circles) on proliferation or IL-2 production of human primary CD4 (A—F) and CD8 T cells (G—L) activated with anti-CD3/CD28 coated beads under strong stimulation (2:1 bead:cell ratio; left panels), intermediate stimulation (1:1 bead:cell ratio; middle panels), or weak stimulation (1:2 bead:cell ratio; right panels) conditions. Cyclosporine A was not tested in CD8 T cells with weak stimulation due to insufficient purified T cells. The assays were performed in 4 independent experiments with samples in duplicates. Data from one representative experiment were presented.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Purification

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Potency and efficacy of Kv1.3 blockers on human T cells under different stimulation conditions.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Inhibition

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Effects of OsK1 and Kv261 on IL-2 production of thapsigargin stimulated CD4 T cells (A) were shown. OsK1, Kv261 peptide and Kv261-HSA-34 were tested in thapsigargin stimulated blood (B) from healthy donors and IL-2 was measured. BTP2 and cyclosporine A were used as reference compounds. The assays were performed twice in CD4 T cells and 3 times in human blood with samples in duplicates. Data from representative experiments were presented.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques:

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Human CD45RO - CCR7 + naïve T cells, CD45RO + CCR7 + central memory (Tcm) and CD45RO + CCR7 - effector memory (Tem) T cells were purified from human blood (A). OsK1 peptide blocked proliferation of the purified human CD4 T cell subsets induced by 2 day stimulation with anti-CD3 in the presence of PBMC (B). BTP2 and cyclosporine A were used as reference compounds in T cell assays. The assays were performed in 8 similar experiments with samples in duplicates. Data from one representative experiment were presented.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: Purification

Journal: PLoS ONE

Article Title: T Cell Subset and Stimulation Strength-Dependent Modulation of T Cell Activation by Kv1.3 Blockers

doi: 10.1371/journal.pone.0170102

Figure Lengend Snippet: Kv1.3 blocker OsK1, Kv261 peptides and Kv261-HSA-34 fusion protein blocked proliferation of CD4 T cells from house dust mite allergic patients following 5 day stimulation with house dust mite extracts (A, B). IL-5 production from house dust mite-specific T cells was inhibited by Kv1.3 blockers as shown with OsK1 and Kv261-HSA-34 (C). OsK1 peptide at 0.3μM inhibited partially the proliferation of PBMC from patients allergic to ragweed after 3 day stimulation with ragweed extracts (D). Decreased proliferative response of individual donor PBMC samples by OsK1 peptide was shown in a separated graph. CTLA4-Ig or cyclosporine A (1μM) were used as reference compounds. The assays in panels A-C were repeated in 4 independent experiments with samples in duplicates. Data from one representative experiment were presented. The study in panel D was done in an experiment with 29 subjects per group. Statistical significance on the difference in proliferation between untreated and OsK1 treated groups were analyzed by 2-way ANOVA and the P value was determined and indicated as *P < 0.05. Readouts in (A) and (C) were performed on CD4 T cells from the same donor whereas those in (B) were from a separate donor. Data from one representative experiment were presented.

Article Snippet: The Kv1.3 channel blockers used in this report were OsK1 peptide (Alomone Labs), ShK peptide (Bachem), Kv261 peptide (Janssen R&D), and Kv261-HSA-34 fusion protein (Janssen R&D).

Techniques: