The random co-polymer glatiramer acetate rapidly kills primary human leukocytes through sialic-acid-dependent cell membrane damage

Stig Hill Christiansen, Xianwei Zhang, Kristian Juul-Madsen, Michael Lykke Hvam, Brian Stougaard Vad, Manja Annette Behrens, Ida Lysgaard Thygesen, Babak Jalilian, Jan Skov Pedersen, Ken Howard, Daniel Otzen, Thomas Vorup-Jensen

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    Abstract

    The formulation glatiramer acetate (GA) is widely used in therapy of multiple sclerosis. GA consists of random copolymers of four amino acids, in ratios that produce a predominantly positive charge and an amphipathic character. With the extraordinary complexity of the drug, several pharmacological modes-of-action were suggested, but so far none, which rationalizes the cationicity and amphipathicity as part of the mode-of-action. Here, we report that GA rapidly kills primary human T lymphocytes and, less actively, monocytes. LL-37 is a cleavage product of human cathelicidin with important roles in innate immunity. It shares the positive charge and amphipathic character of GA, and, as shown here, also the ability to kill human leukocyte. The cytotoxicity of both compounds depends on sialic acid in the cell membrane. The killing was associated with the generation of CD45 + debris, derived from cell membrane deformation. Nanoparticle tracking analysis confirmed the formation of such debris, even at low GA concentrations. Electric cell-substrate impedance sensing measurements also recorded stable alterations in T lymphocytes following such treatment. LL-37 forms oligomers through weak hydrophobic contacts, which is critical for the lytic properties. In our study, SAXS showed that GA also forms this type of contacts. Taken together, our study offers new insight on the immunomodulatory mode-of-action of positively charged co-polymers. The comparison of LL-37 and GA highlights a consistent requirement of certain oligomeric and chemical properties to support cytotoxic effects of cationic polymers targeting human leukocytes.
    Original languageDanish
    JournalBiochimica et Biophysica Acta. Biomembranes
    Volume1859
    Pages (from-to)425–437
    ISSN0005-2736
    DOIs
    Publication statusPublished - 2017

    Keywords

    • Cationic co-polymers
    • Glatiramoids
    • Immunotherapy
    • Cathelicidins

    Cite this

    Christiansen, Stig Hill ; Zhang, Xianwei ; Juul-Madsen, Kristian ; Hvam, Michael Lykke ; Vad, Brian Stougaard ; Behrens, Manja Annette ; Thygesen, Ida Lysgaard ; Jalilian, Babak ; Pedersen, Jan Skov ; Howard, Ken ; Otzen, Daniel ; Vorup-Jensen, Thomas. / The random co-polymer glatiramer acetate rapidly kills primary human leukocytes through sialic-acid-dependent cell membrane damage. In: Biochimica et Biophysica Acta. Biomembranes. 2017 ; Vol. 1859. pp. 425–437.
    @article{682bf468b92a44158944b550c55f5ee2,
    title = "The random co-polymer glatiramer acetate rapidly kills primary human leukocytes through sialic-acid-dependent cell membrane damage",
    abstract = "The formulation glatiramer acetate (GA) is widely used in therapy of multiple sclerosis. GA consists of random copolymers of four amino acids, in ratios that produce a predominantly positive charge and an amphipathic character. With the extraordinary complexity of the drug, several pharmacological modes-of-action were suggested, but so far none, which rationalizes the cationicity and amphipathicity as part of the mode-of-action. Here, we report that GA rapidly kills primary human T lymphocytes and, less actively, monocytes. LL-37 is a cleavage product of human cathelicidin with important roles in innate immunity. It shares the positive charge and amphipathic character of GA, and, as shown here, also the ability to kill human leukocyte. The cytotoxicity of both compounds depends on sialic acid in the cell membrane. The killing was associated with the generation of CD45 + debris, derived from cell membrane deformation. Nanoparticle tracking analysis confirmed the formation of such debris, even at low GA concentrations. Electric cell-substrate impedance sensing measurements also recorded stable alterations in T lymphocytes following such treatment. LL-37 forms oligomers through weak hydrophobic contacts, which is critical for the lytic properties. In our study, SAXS showed that GA also forms this type of contacts. Taken together, our study offers new insight on the immunomodulatory mode-of-action of positively charged co-polymers. The comparison of LL-37 and GA highlights a consistent requirement of certain oligomeric and chemical properties to support cytotoxic effects of cationic polymers targeting human leukocytes.",
    keywords = "Multiple Sclerosis, Cationic co-polymers, Glatiramoids, Immunotherapy, Cathelicidins",
    author = "Christiansen, {Stig Hill} and Xianwei Zhang and Kristian Juul-Madsen and Hvam, {Michael Lykke} and Vad, {Brian Stougaard} and Behrens, {Manja Annette} and Thygesen, {Ida Lysgaard} and Babak Jalilian and Pedersen, {Jan Skov} and Ken Howard and Daniel Otzen and Thomas Vorup-Jensen",
    year = "2017",
    doi = "10.1016/j.bbamem.2017.01.001",
    language = "Dansk",
    volume = "1859",
    pages = "425–437",
    journal = "B B A - Biomembranes",
    issn = "0005-2736",
    publisher = "Elsevier",

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    Christiansen, SH, Zhang, X, Juul-Madsen, K, Hvam, ML, Vad, BS, Behrens, MA, Thygesen, IL, Jalilian, B, Pedersen, JS, Howard, K, Otzen, D & Vorup-Jensen, T 2017, 'The random co-polymer glatiramer acetate rapidly kills primary human leukocytes through sialic-acid-dependent cell membrane damage', Biochimica et Biophysica Acta. Biomembranes, vol. 1859, pp. 425–437. https://doi.org/10.1016/j.bbamem.2017.01.001

    The random co-polymer glatiramer acetate rapidly kills primary human leukocytes through sialic-acid-dependent cell membrane damage. / Christiansen, Stig Hill; Zhang, Xianwei; Juul-Madsen, Kristian; Hvam, Michael Lykke; Vad, Brian Stougaard; Behrens, Manja Annette; Thygesen, Ida Lysgaard; Jalilian, Babak; Pedersen, Jan Skov; Howard, Ken; Otzen, Daniel; Vorup-Jensen, Thomas.

    In: Biochimica et Biophysica Acta. Biomembranes, Vol. 1859, 2017, p. 425–437.

    Research output: Contribution to journalJournal articleResearchpeer-review

    TY - JOUR

    T1 - The random co-polymer glatiramer acetate rapidly kills primary human leukocytes through sialic-acid-dependent cell membrane damage

    AU - Christiansen, Stig Hill

    AU - Zhang, Xianwei

    AU - Juul-Madsen, Kristian

    AU - Hvam, Michael Lykke

    AU - Vad, Brian Stougaard

    AU - Behrens, Manja Annette

    AU - Thygesen, Ida Lysgaard

    AU - Jalilian, Babak

    AU - Pedersen, Jan Skov

    AU - Howard, Ken

    AU - Otzen, Daniel

    AU - Vorup-Jensen, Thomas

    PY - 2017

    Y1 - 2017

    N2 - The formulation glatiramer acetate (GA) is widely used in therapy of multiple sclerosis. GA consists of random copolymers of four amino acids, in ratios that produce a predominantly positive charge and an amphipathic character. With the extraordinary complexity of the drug, several pharmacological modes-of-action were suggested, but so far none, which rationalizes the cationicity and amphipathicity as part of the mode-of-action. Here, we report that GA rapidly kills primary human T lymphocytes and, less actively, monocytes. LL-37 is a cleavage product of human cathelicidin with important roles in innate immunity. It shares the positive charge and amphipathic character of GA, and, as shown here, also the ability to kill human leukocyte. The cytotoxicity of both compounds depends on sialic acid in the cell membrane. The killing was associated with the generation of CD45 + debris, derived from cell membrane deformation. Nanoparticle tracking analysis confirmed the formation of such debris, even at low GA concentrations. Electric cell-substrate impedance sensing measurements also recorded stable alterations in T lymphocytes following such treatment. LL-37 forms oligomers through weak hydrophobic contacts, which is critical for the lytic properties. In our study, SAXS showed that GA also forms this type of contacts. Taken together, our study offers new insight on the immunomodulatory mode-of-action of positively charged co-polymers. The comparison of LL-37 and GA highlights a consistent requirement of certain oligomeric and chemical properties to support cytotoxic effects of cationic polymers targeting human leukocytes.

    AB - The formulation glatiramer acetate (GA) is widely used in therapy of multiple sclerosis. GA consists of random copolymers of four amino acids, in ratios that produce a predominantly positive charge and an amphipathic character. With the extraordinary complexity of the drug, several pharmacological modes-of-action were suggested, but so far none, which rationalizes the cationicity and amphipathicity as part of the mode-of-action. Here, we report that GA rapidly kills primary human T lymphocytes and, less actively, monocytes. LL-37 is a cleavage product of human cathelicidin with important roles in innate immunity. It shares the positive charge and amphipathic character of GA, and, as shown here, also the ability to kill human leukocyte. The cytotoxicity of both compounds depends on sialic acid in the cell membrane. The killing was associated with the generation of CD45 + debris, derived from cell membrane deformation. Nanoparticle tracking analysis confirmed the formation of such debris, even at low GA concentrations. Electric cell-substrate impedance sensing measurements also recorded stable alterations in T lymphocytes following such treatment. LL-37 forms oligomers through weak hydrophobic contacts, which is critical for the lytic properties. In our study, SAXS showed that GA also forms this type of contacts. Taken together, our study offers new insight on the immunomodulatory mode-of-action of positively charged co-polymers. The comparison of LL-37 and GA highlights a consistent requirement of certain oligomeric and chemical properties to support cytotoxic effects of cationic polymers targeting human leukocytes.

    KW - Multiple Sclerosis

    KW - Cationic co-polymers

    KW - Glatiramoids

    KW - Immunotherapy

    KW - Cathelicidins

    U2 - 10.1016/j.bbamem.2017.01.001

    DO - 10.1016/j.bbamem.2017.01.001

    M3 - Tidsskriftartikel

    VL - 1859

    SP - 425

    EP - 437

    JO - B B A - Biomembranes

    JF - B B A - Biomembranes

    SN - 0005-2736

    ER -