Protein engineering of the chemokine CCL20 prevents psoriasiform dermatitis in an IL-23–dependent murine model

Getschman, Anthony E.; Imai, Yasutomo; Larsen, Olav; Peterson, Francis C.; Wu, Xuesong; Rosenkilde, Mette M.; Hwang, Samuel T; Volkman, Brian F.

doi:10.1073/pnas.1704958114

Cited by 51 publications

(64 citation statements)

References 58 publications

(54 reference statements)

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“…CCL20S64C binds CCR6 and induces intracellular calcium release but exhibits minimal chemotactic activity for CCR6+ cells. The administration of CCL20S64C significantly attenuates IL‐23–induced psoriasiform inflammation in mice …”

Section: The Ccl20/ccr6 Axis As Target For the Treatment Of Psoriasismentioning

confidence: 88%

The CCL20 and CCR6 axis in psoriasis

et al. 2019

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Psoriasis is a TNF‐α/IL‐23/IL‐17A–mediated inflammatory skin disease that causes a significant socioeconomic burden in afflicted patients. IL‐17A–producing immune cells, including Th17 cells, are crucial effector cells in the development of psoriasis. IL‐17A stimulates epidermal keratinocytes to produce CCL20, which eventually recruits CCR6 + Th17 cells into the lesional skin. Thus, the CCL20/CCR6 axis works as a driving force that prepares an IL‐17A–rich cutaneous milieu. In this review, we summarize the current research topics on the CCL20/CCR6 axis and the therapeutic intervention of this axis for psoriasis.

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Section: The Ccl20/ccr6 Axis As Target For the Treatment Of Psoriasismentioning

confidence: 88%

The CCL20 and CCR6 axis in psoriasis

et al. 2019

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“…The PathHunter U2OS‐A2 β‐arrestin‐2 cell line (DiscoverX, Fremont, CA, USA) was transfected with CCR6 vector using FuGENE ® 6 approximately 40 h prior to use in β‐arrestin‐2 recruitment assays. Cell lines were maintained as previously described …”

Section: Methodsmentioning

confidence: 99%

“…125 I‐CCL20 WT was produced by oxidative iodination using ChloramineT (PerkinElmer) and purified by reverse‐phase HPLC as previously described . Radioligand displacement assays were performed as previously described . Briefly, CCR6 + COS‐7 cells were seeded at a density of 35,000 cells/well in 96‐well plates.…”

Section: Methodsmentioning

confidence: 99%

“…IP 3 accumulation assays were performed as previously described . Briefly, COS‐7 cells cotransfected with CCR6 and G qi4myr were seeded into 96‐well plates at 35,000 cells/well.…”

Section: Methodsmentioning

confidence: 99%

“…Chemotaxis assays were performed as previously described . Briefly, chemokine dilutions were added to the lower well of HTS Transwell ® 96‐well plates with 5 μm pore size (Corning) and transfected CCR6 + Jurkat cells were added to the upper well at a density of 0.375 × 10 6 cells/mL.…”

Section: Methodsmentioning

confidence: 99%

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Mutational analysis of CCL20 reveals flexibility of N-terminal amino acid composition and length

Riutta

Larsen

Getschman

et al. 2018

Journal of Leukocyte Biology

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Chemokine-chemokine receptor (CKR) interactions are traditionally described by a two-step/two-site mechanism that details the major contact points between chemokine ligands and CKRs leading to ligand recognition and receptor activation. Chemokine recognition site 1 (CRS1) encompasses interactions between the CKR N-terminus and the globular chemokine core. Chemokine recognition site 2 (CRS2) includes interactions between the unstructured chemokine N-terminus and the binding pocket of the receptor. The two-step/two-site paradigm has been an adequate framework to study the intricacies of chemokine:CKR interactions, but emerging studies highlight the limitations of this model. Here, we present studies of CRS2 interactions between the chemokine CCL20 and its cognate receptor CCR6 driven by the hypothesis that CCL20 interacts with CCR6 as described by the two-step/two-site model. CCL20 is a chemokine with an unusually short N-terminus of 5 residues (NH -ASNFD), compared to the average length of 10 residues for chemokine ligands. We have investigated how well CCL20 tolerates manipulation of the N-terminus by monitoring binding affinity of variants and their ability to activate the receptor. We show the CCL20 N-terminus tolerates truncation of up to 3 residues, extension by up to 5 additional residues, and point mutations at 4 of 5 positions with minimal loss of binding affinity and minimal impairment in ability to stimulate calcium mobilization, inositol triphosphate accumulation, chemotaxis, and β-arrestin-2 recruitment. Mutation of the fifth residue, aspartate, to alanine or lysine has a dramatic impact on binding affinity for CCR6 and ligand potency. We postulate CCL20 does not activate CCR6 through the canonical two-step/two-site mechanism of CKR activation.

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Engineered Cytokines for Cancer and Autoimmune Disease Immunotherapy

Uricoli

Birnbaum

et al. 2021

Adv Healthcare Materials

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Cytokine signaling is critical to a range of biological processes including cell development, tissue repair, aging, and immunity. In addition to acting as key signal mediators of the immune system, cytokines can also serve as potent immunotherapies with more than 20 recombinant products currently Food and Drug Administration (FDA)‐approved to treat conditions including hepatitis, multiple sclerosis, arthritis, and various cancers. Yet despite their biological importance and clinical utility, cytokine immunotherapies suffer from intrinsic challenges that limit their therapeutic potential including poor circulation, systemic toxicity, and low tissue‐ or cell‐specificity. In the past decade in particular, methods have been devised to engineer cytokines in order to overcome such challenges and here, the myriad strategies are reviewed that may be employed in order to improve the therapeutic potential of cytokine and chemokine immunotherapies with applications in cancer and autoimmune disease therapy, as well as tissue engineering and regenerative medicine. For clarity, these strategies are collected and presented as they vary across size scales, ranging from single amino acid substitutions, to larger protein‐polymer conjugates, nano/micrometer‐scale particles, and macroscale implants. Together, this work aims to provide readers with a timely view of the field of cytokine engineering with an emphasis on early‐stage therapeutic approaches.

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Protein engineering of the chemokine CCL20 prevents psoriasiform dermatitis in an IL-23–dependent murine model

Cited by 51 publications

References 58 publications

The CCL20 and CCR6 axis in psoriasis

The CCL20 and CCR6 axis in psoriasis

Mutational analysis of CCL20 reveals flexibility of N-terminal amino acid composition and length

Engineered Cytokines for Cancer and Autoimmune Disease Immunotherapy

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