Cholesterol Crystals Activate the Lectin Complement Pathway via Ficolin-2 and Mannose-Binding Lectin: Implications for the Progression of Atherosclerosis

Katrine Pilely; Anne Rosbjerg; Ninette Genster; Péter Gál; Gábor Pál; Bente Halvorsen; Sverre Holm; Pål Aukrust; Siril Skaret Bakke; Bjørnar Sporsheim; Ingunn Nervik; Nathalie Niyonzima; Emil Daniel Bartels; Gregory L Stahl; Tom Eirik Mollnes; Terje Espevik; Peter Garred

doi:10.4049/jimmunol.1502595

Cholesterol Crystals Activate the Lectin Complement Pathway via Ficolin-2 and Mannose-Binding Lectin: Implications for the Progression of Atherosclerosis

Katrine Pilely, Anne Rosbjerg, Ninette Genster, Péter Gál, Gábor Pál, Bente Halvorsen, Sverre Holm, Pål Aukrust, Siril Skaret Bakke, Bjørnar Sporsheim, Ingunn Nervik, Nathalie Niyonzima, Emil Daniel Bartels, Gregory L Stahl, Tom Eirik Mollnes, Terje Espevik, Peter Garred

Department of Clinical Medicine

19 Citations (Scopus)

Abstract

Cholesterol crystals (CC) play an essential role in the formation of atherosclerotic plaques. CC activate the classical and the alternative complement pathways, but the role of the lectin pathway is unknown. We hypothesized that the pattern recognition molecules (PRMs) from the lectin pathway bind CC and function as an upstream innate inflammatory signal in the pathophysiology of atherosclerosis. We investigated the binding of the PRMs mannose-binding lectin (MBL), ficolin-1, ficolin-2, and ficolin-3, the associated serine proteases, and complement activation products to CC in vitro using recombinant proteins, specific inhibitors, as well as deficient and normal sera. Additionally, we examined the deposition of ficolin-2 and MBL in human carotid plaques by immunohistochemistry and fluorescence microscopy. The results showed that the lectin pathway was activated on CC by binding of ficolin-2 and MBL in vitro, resulting in activation and deposition of complement activation products. MBL bound to CC in a calcium-dependent manner whereas ficolin-2 binding was calcium-independent. No binding was observed for ficolin-1 or ficolin-3. MBL and ficolin-2 were present in human carotid plaques, and binding of MBL to CC was confirmed in vivo by immunohistochemistry, showing localization of MBL around CC clefts. Moreover, we demonstrated that IgM, but not IgG, bound to CC in vitro and that C1q binding was facilitated by IgM. In conclusion, our study demonstrates that PRMs from the lectin pathway recognize CC and provides evidence for an important role for this pathway in the inflammatory response induced by CC in the pathophysiology of atherosclerosis.

Original language	English
Journal	Journal of immunology (Baltimore, Md. : 1950)
Volume	196
Issue number	12
Pages (from-to)	5064-5074
Number of pages	11
ISSN	0022-1767
DOIs	https://doi.org/10.4049/jimmunol.1502595
Publication status	Published - 15 Jun 2016

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Pilely, K., Rosbjerg, A., Genster, N., Gál, P., Pál, G., Halvorsen, B., Holm, S., Aukrust, P., Bakke, S. S., Sporsheim, B., Nervik, I., Niyonzima, N., Bartels, E. D., Stahl, G. L., Mollnes, T. E., Espevik, T., & Garred, P. (2016). Cholesterol Crystals Activate the Lectin Complement Pathway via Ficolin-2 and Mannose-Binding Lectin: Implications for the Progression of Atherosclerosis. Journal of immunology (Baltimore, Md. : 1950), 196(12), 5064-5074. https://doi.org/10.4049/jimmunol.1502595

Cholesterol Crystals Activate the Lectin Complement Pathway via Ficolin-2 and Mannose-Binding Lectin: Implications for the Progression of Atherosclerosis. / Pilely, Katrine; Rosbjerg, Anne; Genster, Ninette et al.
In: Journal of immunology (Baltimore, Md. : 1950), Vol. 196, No. 12, 15.06.2016, p. 5064-5074.

Research output: Contribution to journal › Journal article › Research › peer-review

Pilely, K, Rosbjerg, A, Genster, N, Gál, P, Pál, G, Halvorsen, B, Holm, S, Aukrust, P, Bakke, SS, Sporsheim, B, Nervik, I, Niyonzima, N, Bartels, ED, Stahl, GL, Mollnes, TE, Espevik, T & Garred, P 2016, 'Cholesterol Crystals Activate the Lectin Complement Pathway via Ficolin-2 and Mannose-Binding Lectin: Implications for the Progression of Atherosclerosis', Journal of immunology (Baltimore, Md. : 1950), vol. 196, no. 12, pp. 5064-5074. https://doi.org/10.4049/jimmunol.1502595

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title = "Cholesterol Crystals Activate the Lectin Complement Pathway via Ficolin-2 and Mannose-Binding Lectin: Implications for the Progression of Atherosclerosis",

abstract = "Cholesterol crystals (CC) play an essential role in the formation of atherosclerotic plaques. CC activate the classical and the alternative complement pathways, but the role of the lectin pathway is unknown. We hypothesized that the pattern recognition molecules (PRMs) from the lectin pathway bind CC and function as an upstream innate inflammatory signal in the pathophysiology of atherosclerosis. We investigated the binding of the PRMs mannose-binding lectin (MBL), ficolin-1, ficolin-2, and ficolin-3, the associated serine proteases, and complement activation products to CC in vitro using recombinant proteins, specific inhibitors, as well as deficient and normal sera. Additionally, we examined the deposition of ficolin-2 and MBL in human carotid plaques by immunohistochemistry and fluorescence microscopy. The results showed that the lectin pathway was activated on CC by binding of ficolin-2 and MBL in vitro, resulting in activation and deposition of complement activation products. MBL bound to CC in a calcium-dependent manner whereas ficolin-2 binding was calcium-independent. No binding was observed for ficolin-1 or ficolin-3. MBL and ficolin-2 were present in human carotid plaques, and binding of MBL to CC was confirmed in vivo by immunohistochemistry, showing localization of MBL around CC clefts. Moreover, we demonstrated that IgM, but not IgG, bound to CC in vitro and that C1q binding was facilitated by IgM. In conclusion, our study demonstrates that PRMs from the lectin pathway recognize CC and provides evidence for an important role for this pathway in the inflammatory response induced by CC in the pathophysiology of atherosclerosis.",

author = "Katrine Pilely and Anne Rosbjerg and Ninette Genster and P{\'e}ter G{\'a}l and G{\'a}bor P{\'a}l and Bente Halvorsen and Sverre Holm and P{\aa}l Aukrust and Bakke, {Siril Skaret} and Bj{\o}rnar Sporsheim and Ingunn Nervik and Nathalie Niyonzima and Bartels, {Emil Daniel} and Stahl, {Gregory L} and Mollnes, {Tom Eirik} and Terje Espevik and Peter Garred",

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doi = "10.4049/jimmunol.1502595",

language = "English",

volume = "196",

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TY - JOUR

T1 - Cholesterol Crystals Activate the Lectin Complement Pathway via Ficolin-2 and Mannose-Binding Lectin

T2 - Implications for the Progression of Atherosclerosis

AU - Pilely, Katrine

AU - Rosbjerg, Anne

AU - Genster, Ninette

AU - Gál, Péter

AU - Pál, Gábor

AU - Halvorsen, Bente

AU - Holm, Sverre

AU - Aukrust, Pål

AU - Bakke, Siril Skaret

AU - Sporsheim, Bjørnar

AU - Nervik, Ingunn

AU - Niyonzima, Nathalie

AU - Bartels, Emil Daniel

AU - Stahl, Gregory L

AU - Mollnes, Tom Eirik

AU - Espevik, Terje

AU - Garred, Peter

PY - 2016/6/15

Y1 - 2016/6/15

N2 - Cholesterol crystals (CC) play an essential role in the formation of atherosclerotic plaques. CC activate the classical and the alternative complement pathways, but the role of the lectin pathway is unknown. We hypothesized that the pattern recognition molecules (PRMs) from the lectin pathway bind CC and function as an upstream innate inflammatory signal in the pathophysiology of atherosclerosis. We investigated the binding of the PRMs mannose-binding lectin (MBL), ficolin-1, ficolin-2, and ficolin-3, the associated serine proteases, and complement activation products to CC in vitro using recombinant proteins, specific inhibitors, as well as deficient and normal sera. Additionally, we examined the deposition of ficolin-2 and MBL in human carotid plaques by immunohistochemistry and fluorescence microscopy. The results showed that the lectin pathway was activated on CC by binding of ficolin-2 and MBL in vitro, resulting in activation and deposition of complement activation products. MBL bound to CC in a calcium-dependent manner whereas ficolin-2 binding was calcium-independent. No binding was observed for ficolin-1 or ficolin-3. MBL and ficolin-2 were present in human carotid plaques, and binding of MBL to CC was confirmed in vivo by immunohistochemistry, showing localization of MBL around CC clefts. Moreover, we demonstrated that IgM, but not IgG, bound to CC in vitro and that C1q binding was facilitated by IgM. In conclusion, our study demonstrates that PRMs from the lectin pathway recognize CC and provides evidence for an important role for this pathway in the inflammatory response induced by CC in the pathophysiology of atherosclerosis.

AB - Cholesterol crystals (CC) play an essential role in the formation of atherosclerotic plaques. CC activate the classical and the alternative complement pathways, but the role of the lectin pathway is unknown. We hypothesized that the pattern recognition molecules (PRMs) from the lectin pathway bind CC and function as an upstream innate inflammatory signal in the pathophysiology of atherosclerosis. We investigated the binding of the PRMs mannose-binding lectin (MBL), ficolin-1, ficolin-2, and ficolin-3, the associated serine proteases, and complement activation products to CC in vitro using recombinant proteins, specific inhibitors, as well as deficient and normal sera. Additionally, we examined the deposition of ficolin-2 and MBL in human carotid plaques by immunohistochemistry and fluorescence microscopy. The results showed that the lectin pathway was activated on CC by binding of ficolin-2 and MBL in vitro, resulting in activation and deposition of complement activation products. MBL bound to CC in a calcium-dependent manner whereas ficolin-2 binding was calcium-independent. No binding was observed for ficolin-1 or ficolin-3. MBL and ficolin-2 were present in human carotid plaques, and binding of MBL to CC was confirmed in vivo by immunohistochemistry, showing localization of MBL around CC clefts. Moreover, we demonstrated that IgM, but not IgG, bound to CC in vitro and that C1q binding was facilitated by IgM. In conclusion, our study demonstrates that PRMs from the lectin pathway recognize CC and provides evidence for an important role for this pathway in the inflammatory response induced by CC in the pathophysiology of atherosclerosis.

U2 - 10.4049/jimmunol.1502595

DO - 10.4049/jimmunol.1502595

M3 - Journal article

C2 - 27183610

SN - 0022-1767

VL - 196

SP - 5064

EP - 5074

JO - Journal of immunology (Baltimore, Md. : 1950)

JF - Journal of immunology (Baltimore, Md. : 1950)

IS - 12

ER -

Cholesterol Crystals Activate the Lectin Complement Pathway via Ficolin-2 and Mannose-Binding Lectin: Implications for the Progression of Atherosclerosis

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