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Fully human antagonistic antibodies against CCR4 potently inhibit cell signaling and chemotaxis.

Hagemann UB, Gunnarsson L, Géraudie S, Scheffler U, Griep RA, Reiersen H, Duncan AR, Kiprijanov SM - PLoS ONE (2014)

Bottom Line: In a mouse model of human T-cell lymphoma, significant survival benefit was demonstrated for animals treated with the newly selected anti-CCR4 antibodies.For the first time, successful generation of anti- G-protein coupled chemokine receptor (GPCR) antibodies using human non-immune library and phage display on GPCR-expressing cells was demonstrated.The data demonstrate that the anti-tumor activity in vivo is mediated, at least in part, through Fc-receptor dependent effector mechanisms, such as ADCC and phagocytosis.

View Article: PubMed Central - PubMed

Affiliation: Affitech Research AS, Oslo, Norway.

ABSTRACT

Background: CC chemokine receptor 4 (CCR4) represents a potentially important target for cancer immunotherapy due to its expression on tumor infiltrating immune cells including regulatory T cells (Tregs) and on tumor cells in several cancer types and its role in metastasis.

Methodology: Using phage display, human antibody library, affinity maturation and a cell-based antibody selection strategy, the antibody variants against human CCR4 were generated. These antibodies effectively competed with ligand binding, were able to block ligand-induced signaling and cell migration, and demonstrated efficient killing of CCR4-positive tumor cells via ADCC and phagocytosis. In a mouse model of human T-cell lymphoma, significant survival benefit was demonstrated for animals treated with the newly selected anti-CCR4 antibodies.

Significance: For the first time, successful generation of anti- G-protein coupled chemokine receptor (GPCR) antibodies using human non-immune library and phage display on GPCR-expressing cells was demonstrated. The generated anti-CCR4 antibodies possess a dual mode of action (inhibition of ligand-induced signaling and antibody-directed tumor cell killing). The data demonstrate that the anti-tumor activity in vivo is mediated, at least in part, through Fc-receptor dependent effector mechanisms, such as ADCC and phagocytosis. Anti-CC chemokine receptor 4 antibodies inhibiting receptor signaling have potential as immunomodulatory antibodies for cancer.

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Related in: MedlinePlus

Effect of human anti-CCR4 antibody 9E10J on platelet aggregation.(a) Binding of 9E10J IgG1 at concentration 10 µg/mL to human platelets isolated from the fresh donor blood. As negative controls, binding histograms for an isotype control antibody (Isotype) and of a secondary antibody alone (anti-human-PE) are shown. (b) Effect of anti-CCR4 antibody 9E10 and of the isotype control antibody on platelet aggregation in comparison with ADP. (c) Platelet aggregation induced by CCR4 ligands, CCL17 and CCL22, in comparison with ADP. (d, e) Ligand-induced aggregation of platelets pre-incubated with either the isotype control antibody (d) or anti-CCR4 antibody 9E10J (e).
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pone-0103776-g007: Effect of human anti-CCR4 antibody 9E10J on platelet aggregation.(a) Binding of 9E10J IgG1 at concentration 10 µg/mL to human platelets isolated from the fresh donor blood. As negative controls, binding histograms for an isotype control antibody (Isotype) and of a secondary antibody alone (anti-human-PE) are shown. (b) Effect of anti-CCR4 antibody 9E10 and of the isotype control antibody on platelet aggregation in comparison with ADP. (c) Platelet aggregation induced by CCR4 ligands, CCL17 and CCL22, in comparison with ADP. (d, e) Ligand-induced aggregation of platelets pre-incubated with either the isotype control antibody (d) or anti-CCR4 antibody 9E10J (e).

Mentions: Expression of functional CCR4 on human platelets [43] raised the question whether the isolated human antibodies would bind to platelets and affect their activation. Experiments performed with an affinity-matured variant 9E10J demonstrated that the antibody indeed binds to platelets (Figure 7a), however, this binding did not lead to platelet activation nor aggregation (Figure 7b). In contrast, the CCR4 ligands when incubated with platelets strongly induced their aggregation (Figure 7c). This effect of ligands was not changed in presence of the isotype control antibody (Figure 7d) and was only slightly influenced by the anti-CCR4 antibody 9E10J at a concentration used in the assay (Figure 7e).


Fully human antagonistic antibodies against CCR4 potently inhibit cell signaling and chemotaxis.

Hagemann UB, Gunnarsson L, Géraudie S, Scheffler U, Griep RA, Reiersen H, Duncan AR, Kiprijanov SM - PLoS ONE (2014)

Effect of human anti-CCR4 antibody 9E10J on platelet aggregation.(a) Binding of 9E10J IgG1 at concentration 10 µg/mL to human platelets isolated from the fresh donor blood. As negative controls, binding histograms for an isotype control antibody (Isotype) and of a secondary antibody alone (anti-human-PE) are shown. (b) Effect of anti-CCR4 antibody 9E10 and of the isotype control antibody on platelet aggregation in comparison with ADP. (c) Platelet aggregation induced by CCR4 ligands, CCL17 and CCL22, in comparison with ADP. (d, e) Ligand-induced aggregation of platelets pre-incubated with either the isotype control antibody (d) or anti-CCR4 antibody 9E10J (e).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4117600&req=5

pone-0103776-g007: Effect of human anti-CCR4 antibody 9E10J on platelet aggregation.(a) Binding of 9E10J IgG1 at concentration 10 µg/mL to human platelets isolated from the fresh donor blood. As negative controls, binding histograms for an isotype control antibody (Isotype) and of a secondary antibody alone (anti-human-PE) are shown. (b) Effect of anti-CCR4 antibody 9E10 and of the isotype control antibody on platelet aggregation in comparison with ADP. (c) Platelet aggregation induced by CCR4 ligands, CCL17 and CCL22, in comparison with ADP. (d, e) Ligand-induced aggregation of platelets pre-incubated with either the isotype control antibody (d) or anti-CCR4 antibody 9E10J (e).
Mentions: Expression of functional CCR4 on human platelets [43] raised the question whether the isolated human antibodies would bind to platelets and affect their activation. Experiments performed with an affinity-matured variant 9E10J demonstrated that the antibody indeed binds to platelets (Figure 7a), however, this binding did not lead to platelet activation nor aggregation (Figure 7b). In contrast, the CCR4 ligands when incubated with platelets strongly induced their aggregation (Figure 7c). This effect of ligands was not changed in presence of the isotype control antibody (Figure 7d) and was only slightly influenced by the anti-CCR4 antibody 9E10J at a concentration used in the assay (Figure 7e).

Bottom Line: In a mouse model of human T-cell lymphoma, significant survival benefit was demonstrated for animals treated with the newly selected anti-CCR4 antibodies.For the first time, successful generation of anti- G-protein coupled chemokine receptor (GPCR) antibodies using human non-immune library and phage display on GPCR-expressing cells was demonstrated.The data demonstrate that the anti-tumor activity in vivo is mediated, at least in part, through Fc-receptor dependent effector mechanisms, such as ADCC and phagocytosis.

View Article: PubMed Central - PubMed

Affiliation: Affitech Research AS, Oslo, Norway.

ABSTRACT

Background: CC chemokine receptor 4 (CCR4) represents a potentially important target for cancer immunotherapy due to its expression on tumor infiltrating immune cells including regulatory T cells (Tregs) and on tumor cells in several cancer types and its role in metastasis.

Methodology: Using phage display, human antibody library, affinity maturation and a cell-based antibody selection strategy, the antibody variants against human CCR4 were generated. These antibodies effectively competed with ligand binding, were able to block ligand-induced signaling and cell migration, and demonstrated efficient killing of CCR4-positive tumor cells via ADCC and phagocytosis. In a mouse model of human T-cell lymphoma, significant survival benefit was demonstrated for animals treated with the newly selected anti-CCR4 antibodies.

Significance: For the first time, successful generation of anti- G-protein coupled chemokine receptor (GPCR) antibodies using human non-immune library and phage display on GPCR-expressing cells was demonstrated. The generated anti-CCR4 antibodies possess a dual mode of action (inhibition of ligand-induced signaling and antibody-directed tumor cell killing). The data demonstrate that the anti-tumor activity in vivo is mediated, at least in part, through Fc-receptor dependent effector mechanisms, such as ADCC and phagocytosis. Anti-CC chemokine receptor 4 antibodies inhibiting receptor signaling have potential as immunomodulatory antibodies for cancer.

Show MeSH
Related in: MedlinePlus