A Novel Biological Role for Peptidyl-Arginine Deiminases: Citrullination of Cathelicidin LL-37 Controls the Immunostimulatory Potential of Cell-Free DNA

Alicia Wong; Danuta Bryzek; Ewelina Dobosz; Carsten Scavenius; Pavel Svoboda; Maria Rapala-Kozik; Adam Lesner; Ivo Frydrych; Jan Enghild; Piotr Mydel; Jan Pohl; Paul R Thompson; Jan Potempa; Joanna Koziel

doi:10.4049/jimmunol.1701391

A Novel Biological Role for Peptidyl-Arginine Deiminases: Citrullination of Cathelicidin LL-37 Controls the Immunostimulatory Potential of Cell-Free DNA

Alicia Wong, Danuta Bryzek, Ewelina Dobosz, Carsten Scavenius, Pavel Svoboda, Maria Rapala-Kozik, Adam Lesner, Ivo Frydrych, Jan Enghild, Piotr Mydel, Jan Pohl, Paul R Thompson, Jan Potempa, Joanna Koziel

Interdisciplinary Nanoscience Center

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

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Abstract

LL-37, the only human cathelicidin that is released during inflammation, is a potent regulator of immune responses by facilitating delivery of oligonucleotides to intracellular TLR-9, thereby enhancing the response of human plasmacytoid dendritic cells (pDCs) to extracellular DNA. Although important for pathogen recognition, this mechanism may facilitate development of autoimmune diseases. In this article, we show that citrullination of LL-37 by peptidyl-arginine deiminases (PADs) hindered peptide-dependent DNA uptake and sensing by pDCs. In contrast, carbamylation of the peptide (homocitrullination of Lys residues) had no effect. The efficiency of LL-37 binding to oligonucleotides and activation of pDCs was found to be inversely proportional to the number of citrullinated residues in the peptide. Similarly, preincubation of carbamylated LL-37 with PAD2 abrogated the peptide's ability to bind DNA. Conversely, LL-37 with Arg residues substituted by homoarginine, which cannot be deiminated, elicited full activity of native LL-37 regardless of PAD2 treatment. Taken together, the data showed that citrullination abolished LL-37 ability to bind DNA and altered the immunomodulatory function of the peptide. Both activities were dependent on the proper distribution of guanidinium side chains in the native peptide sequence. Moreover, our data suggest that cathelicidin/LL-37 is citrullinated by PADs during NET formation, thus affecting the inflammatory potential of NETs. Together this may represent a novel mechanism for preventing the breakdown of immunotolerance, which is dependent on the response of APCs to self-molecules (including cell-free DNA); overactivation may facilitate development of autoimmunity.

Original language	English
Journal	Journal of Immunology
Volume	200
Issue	7
Pages (from-to)	2327-2340
Number of pages	14
ISSN	0022-1767
DOIs	https://doi.org/10.4049/jimmunol.1701391
Publication status	Published - 1 Apr 2018

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Journal Article

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https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5860981/pdf/nihms938212.pdf

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Wong, A., Bryzek, D., Dobosz, E., Scavenius, C., Svoboda, P., Rapala-Kozik, M., Lesner, A., Frydrych, I., Enghild, J., Mydel, P., Pohl, J., Thompson, P. R., Potempa, J., & Koziel, J. (2018). A Novel Biological Role for Peptidyl-Arginine Deiminases: Citrullination of Cathelicidin LL-37 Controls the Immunostimulatory Potential of Cell-Free DNA. Journal of Immunology, 200(7), 2327-2340. https://doi.org/10.4049/jimmunol.1701391

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title = "A Novel Biological Role for Peptidyl-Arginine Deiminases: Citrullination of Cathelicidin LL-37 Controls the Immunostimulatory Potential of Cell-Free DNA",

abstract = "LL-37, the only human cathelicidin that is released during inflammation, is a potent regulator of immune responses by facilitating delivery of oligonucleotides to intracellular TLR-9, thereby enhancing the response of human plasmacytoid dendritic cells (pDCs) to extracellular DNA. Although important for pathogen recognition, this mechanism may facilitate development of autoimmune diseases. In this article, we show that citrullination of LL-37 by peptidyl-arginine deiminases (PADs) hindered peptide-dependent DNA uptake and sensing by pDCs. In contrast, carbamylation of the peptide (homocitrullination of Lys residues) had no effect. The efficiency of LL-37 binding to oligonucleotides and activation of pDCs was found to be inversely proportional to the number of citrullinated residues in the peptide. Similarly, preincubation of carbamylated LL-37 with PAD2 abrogated the peptide's ability to bind DNA. Conversely, LL-37 with Arg residues substituted by homoarginine, which cannot be deiminated, elicited full activity of native LL-37 regardless of PAD2 treatment. Taken together, the data showed that citrullination abolished LL-37 ability to bind DNA and altered the immunomodulatory function of the peptide. Both activities were dependent on the proper distribution of guanidinium side chains in the native peptide sequence. Moreover, our data suggest that cathelicidin/LL-37 is citrullinated by PADs during NET formation, thus affecting the inflammatory potential of NETs. Together this may represent a novel mechanism for preventing the breakdown of immunotolerance, which is dependent on the response of APCs to self-molecules (including cell-free DNA); overactivation may facilitate development of autoimmunity.",

keywords = "Journal Article",

author = "Alicia Wong and Danuta Bryzek and Ewelina Dobosz and Carsten Scavenius and Pavel Svoboda and Maria Rapala-Kozik and Adam Lesner and Ivo Frydrych and Jan Enghild and Piotr Mydel and Jan Pohl and Thompson, {Paul R} and Jan Potempa and Joanna Koziel",

note = "Copyright {\textcopyright} 2018 by The American Association of Immunologists, Inc.",

year = "2018",

month = apr,

day = "1",

doi = "10.4049/jimmunol.1701391",

language = "English",

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Wong, A, Bryzek, D, Dobosz, E, Scavenius, C, Svoboda, P, Rapala-Kozik, M, Lesner, A, Frydrych, I, Enghild, J, Mydel, P, Pohl, J, Thompson, PR, Potempa, J & Koziel, J 2018, 'A Novel Biological Role for Peptidyl-Arginine Deiminases: Citrullination of Cathelicidin LL-37 Controls the Immunostimulatory Potential of Cell-Free DNA', Journal of Immunology, vol. 200, no. 7, pp. 2327-2340. https://doi.org/10.4049/jimmunol.1701391

A Novel Biological Role for Peptidyl-Arginine Deiminases: Citrullination of Cathelicidin LL-37 Controls the Immunostimulatory Potential of Cell-Free DNA. / Wong, Alicia; Bryzek, Danuta; Dobosz, Ewelina et al.
In: Journal of Immunology, Vol. 200, No. 7, 01.04.2018, p. 2327-2340.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - A Novel Biological Role for Peptidyl-Arginine Deiminases

T2 - Citrullination of Cathelicidin LL-37 Controls the Immunostimulatory Potential of Cell-Free DNA

AU - Wong, Alicia

AU - Bryzek, Danuta

AU - Dobosz, Ewelina

AU - Scavenius, Carsten

AU - Svoboda, Pavel

AU - Rapala-Kozik, Maria

AU - Lesner, Adam

AU - Frydrych, Ivo

AU - Enghild, Jan

AU - Mydel, Piotr

AU - Pohl, Jan

AU - Thompson, Paul R

AU - Potempa, Jan

AU - Koziel, Joanna

PY - 2018/4/1

Y1 - 2018/4/1

N2 - LL-37, the only human cathelicidin that is released during inflammation, is a potent regulator of immune responses by facilitating delivery of oligonucleotides to intracellular TLR-9, thereby enhancing the response of human plasmacytoid dendritic cells (pDCs) to extracellular DNA. Although important for pathogen recognition, this mechanism may facilitate development of autoimmune diseases. In this article, we show that citrullination of LL-37 by peptidyl-arginine deiminases (PADs) hindered peptide-dependent DNA uptake and sensing by pDCs. In contrast, carbamylation of the peptide (homocitrullination of Lys residues) had no effect. The efficiency of LL-37 binding to oligonucleotides and activation of pDCs was found to be inversely proportional to the number of citrullinated residues in the peptide. Similarly, preincubation of carbamylated LL-37 with PAD2 abrogated the peptide's ability to bind DNA. Conversely, LL-37 with Arg residues substituted by homoarginine, which cannot be deiminated, elicited full activity of native LL-37 regardless of PAD2 treatment. Taken together, the data showed that citrullination abolished LL-37 ability to bind DNA and altered the immunomodulatory function of the peptide. Both activities were dependent on the proper distribution of guanidinium side chains in the native peptide sequence. Moreover, our data suggest that cathelicidin/LL-37 is citrullinated by PADs during NET formation, thus affecting the inflammatory potential of NETs. Together this may represent a novel mechanism for preventing the breakdown of immunotolerance, which is dependent on the response of APCs to self-molecules (including cell-free DNA); overactivation may facilitate development of autoimmunity.

AB - LL-37, the only human cathelicidin that is released during inflammation, is a potent regulator of immune responses by facilitating delivery of oligonucleotides to intracellular TLR-9, thereby enhancing the response of human plasmacytoid dendritic cells (pDCs) to extracellular DNA. Although important for pathogen recognition, this mechanism may facilitate development of autoimmune diseases. In this article, we show that citrullination of LL-37 by peptidyl-arginine deiminases (PADs) hindered peptide-dependent DNA uptake and sensing by pDCs. In contrast, carbamylation of the peptide (homocitrullination of Lys residues) had no effect. The efficiency of LL-37 binding to oligonucleotides and activation of pDCs was found to be inversely proportional to the number of citrullinated residues in the peptide. Similarly, preincubation of carbamylated LL-37 with PAD2 abrogated the peptide's ability to bind DNA. Conversely, LL-37 with Arg residues substituted by homoarginine, which cannot be deiminated, elicited full activity of native LL-37 regardless of PAD2 treatment. Taken together, the data showed that citrullination abolished LL-37 ability to bind DNA and altered the immunomodulatory function of the peptide. Both activities were dependent on the proper distribution of guanidinium side chains in the native peptide sequence. Moreover, our data suggest that cathelicidin/LL-37 is citrullinated by PADs during NET formation, thus affecting the inflammatory potential of NETs. Together this may represent a novel mechanism for preventing the breakdown of immunotolerance, which is dependent on the response of APCs to self-molecules (including cell-free DNA); overactivation may facilitate development of autoimmunity.

KW - Journal Article

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U2 - 10.4049/jimmunol.1701391

DO - 10.4049/jimmunol.1701391

M3 - Journal article

C2 - 29475987

SN - 0022-1767

VL - 200

SP - 2327

EP - 2340

JO - Journal of Immunology

JF - Journal of Immunology

IS - 7

ER -

A Novel Biological Role for Peptidyl-Arginine Deiminases: Citrullination of Cathelicidin LL-37 Controls the Immunostimulatory Potential of Cell-Free DNA

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