Functional Analysis of Ficolin-3 Mediated Complement Activation

Hein, Estrid; Honoré, Christian; Skjoedt, Mikkel‐Ole; Munthe-Fog, Lea; Hummelshøj, Tina; Garred, Peter

doi:10.1371/journal.pone.0015443

Cited by 61 publications

(60 citation statements)

References 42 publications

(70 reference statements)

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“…Binding of ficolin-1, -2, or -3 was detected using 5 mg/ml mouse anti-human ficolin-1 FCN106, 0.5 mg/ml (recombinant protein) or 5 mg/ml (serum) mouse anti-human ficolin-2 mAb FCN219, or 5 mg/ml mouse anti-human ficolin-3 mAb FCN334 (30 min at 4˚C), followed by 2.5 mg/ml FITC-conjugated polyclonal goat anti-mouse Ab (20 min at 4˚C). In some experiments, CC were incubated with serum preincubated 5 min at RT with 10 mg/ml ficolin-2 inhibitory mAb FCN212, 10 mg/ml AcBSA prepared as described in Hein et al (46), 10 mg/ml BSA, 10 mM EDTA, 10 mM EGTA/Mg, 20 mM GlcNAc, 20 mM D-mannose, or 20 mM D-(+)-galactose. Binding of MASPs/MAP-1 to CC.…”

Section: Collection Of Serum Samplesmentioning

confidence: 99%

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

Pilely

Rosbjerg

Genster

et al. 2016

The Journal of Immunology

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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.

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Section: Collection Of Serum Samplesmentioning

confidence: 99%

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

Pilely

Rosbjerg

Genster

et al. 2016

The Journal of Immunology

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“…M7504) or 5 mg/ml acetylated BSA [AcBSA; as described by Hein et al (22)] in PBS overnight at 4˚C. Plates were washed three times in TBST/Ca 2+ washing/ dilution buffer (10 mM Tris, 145 mM NaCl, 2 mM CaCl 2 , 0.05% Tween-20).…”

Section: C1-inh Interaction With Ligand-bound Mbl/ficolin-3/masp Compmentioning

confidence: 99%

The Levels of the Lectin Pathway Serine Protease MASP-1 and Its Complex Formation with C1 Inhibitor Are Linked to the Severity of Hereditary Angioedema

Hansen

Csuka

Munthe-Fog

et al. 2015

The Journal of Immunology

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C1 inhibitor (C1-INH) is known to form complexes with the lectin complement pathway serine proteases MASP-1 and MASP-2. Deficiency of C1-INH is associated with hereditary angioedema (HAE), an autosomal inherited disease characterized by swelling attacks caused by elevated levels of bradykinin. MASP-1 was shown to cleave high m.w. kininogen into bradykinin; therefore, we hypothesized that MASP-1 levels and the quantity of MASP-1/C1-INH complexes might be associated with different paraclinical and clinical outcomes of HAE. We measured MASP-1 serum concentrations and endogenous MASP-1/C1-INH complex levels in 128 HAE patients and 100 controls. Relatively high levels of pre-existing MASP-1/C1-INH complexes were observed in normal serum, and we found that both the serum levels of MASP-1 and the complex formation between MASP-1 and C1-INH were significantly reduced in HAE patients compared with matched controls (p < 0.0001). The level of MASP-1 and MASP-1/C1-INH complexes in HE patients correlated with the level of C1-INH (p = 0.0009 and p = 0.0047, respectively), the level of C4 (p = 0.0084 and p < 0.0001, respectively), and the number of attacks in the year of blood sampling (p = 0.0075 and p = 0.0058, respectively). In conclusion, we show that MASP-1/C1-INH complexes circulate in normal human blood. The levels of MASP-1 and MASP-1/C1-INH complexes are reduced in HAE patients compared with controls. Both MASP-1 and MASP-1/C1-INH complexes are related to the degree of complement C4 consumption, as well as the severity of disease. These results suggest that MASP-1 may exert a previously unrecognized role in the pathophysiology of HAE.

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“…24) in PBS. Between each step of the procedure, the plates were washed four times with 50 mM Tris-HCl, 150 mM NaCl, and 0.1% Tween 20 (pH 7.5).…”

Section: Complement Activation Assaysmentioning

confidence: 99%

A metalloproteinase mirolysin of Tannerella forsythia inhibits all pathways of the complement system

Jusko

Potempa

Książek

et al. 2013

Molecular Immunology

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Functional Analysis of Ficolin-3 Mediated Complement Activation

Cited by 61 publications

References 42 publications

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

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

The Levels of the Lectin Pathway Serine Protease MASP-1 and Its Complex Formation with C1 Inhibitor Are Linked to the Severity of Hereditary Angioedema

A metalloproteinase mirolysin of Tannerella forsythia inhibits all pathways of the complement system

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