Low ficolin-3 levels in early follow-up serum samples are associated with the severity and unfavorable outcome of acute ischemic stroke

Füst George, Munthe-Fog Lea, Illes Zsolt, Széplaki Gábor, Molnar Tihamér, Pusch Gabriella, Hirschberg Kristóf, Szegedi Robert, Széplaki Zoltán, Prohászka Zoltán, Skjoedt Mikkel-Ole, Garred Peter

Description

A number of data indicate that the lectin pathway of complement activation contributes to the pathophysiology of ischemic stroke. The lectin pathway may be triggered by the binding of mannose-binding lectin (MBL), ficolin-2 or ficolin-3 to different ligands. Although several papers demonstrated the significance of MBL in ischemic stroke, the role of ficolins has not been examined. Methods Sera were obtained within 12 hours after the onset of ischemic stroke (admission samples) and 3-4 days later (follow-up samples) from 65 patients. The control group comprised 100 healthy individuals and 135 patients with significant carotid stenosis (patient controls). The concentrations of ficolin-2 and ficolin-3, initiator molecules of the lectin complement pathway, were measured by ELISA methods. Concentration of C-reactive protein (CRP) was also determined by a particle-enhanced immunturbidimetric assay. Results Concentrations of both ficolin-2 and ficolin-3 were significantly (p < 0.001) decreased in both the admission and in the follow-up samples of patients with definite ischemic stroke as compared to healthy subjects. Concentrations of ficolin-2 and ficolin-3 were even higher in patient controls than in healthy subjects, indicating that the decreased levels in sera during the acute phase of stroke are related to the acute ischemic event. Ficolin-3 levels in the follow-up samples inversely correlated with the severity of stroke indicated by NIH scale on admission. In follow-up samples an inverse correlation was observed between ficolin-3 levels and concentration of S100β, an indicator of the size of cerebral infarct. Patients with low ficolin-3 levels and high CRP levels in the follow up samples had a significantly worse outcome (adjusted ORs 5.6 and 3.9, respectively) as measured by the modified Rankin scale compared to patients with higher ficolin-3 and lower CRP concentrations. High CRP concentrations were similarly predictive for worse outcome, and the effects of low ficolin-3 and high CRP were independent. Conclusions Our findings indicate that ficolin-mediated lectin pathways of complement activation contribute to the pathogenesis of ischemic stroke and may be additive to complement-independent inflammatory processes.

Sujets

• Stroke
• Outcome
• Complément
• Mannan-binding lectin pathway
• CRP

Informations

• Publié par: biomed
• Publié le: 01 janvier 2011
• Nombre de lectures: 10
• Langue: English
• Poids de l'ouvrage: 1 Mo

Extrait

Füst
etal
.
JournalofNeuroinfla
m
ation
2011,
8
:185
http://www.jneuroinflammation.com/content/8/1/185

RESEARCH

JONUERUNRAIOL NOFL FAMATIONOpenAccess

Lowficolin-3levelsinearlyfollow-upserum
samplesareassociatedwiththeseverityand
unfavorableoutcomeofacuteischemicstroke
GeorgeFüst
1*
,LeaMunthe-Fog
2
,ZsoltIlles
3
,GáborSzéplaki
1
,TihamérMolnar
4
,GabriellaPusch
3
,
KristófHirschberg
5,7
,RobertSzegedi
6
,ZoltánSzéplaki
6
,ZoltánProhászka
1
,Mikkel-OleSkjoedt
2
andPeterGarred
2

Abstract
Background:
Anumberofdataindicatethatthelectinpathwayofcomplementactivationcontributestothe
pathophysiologyofischemicstroke.Thelectinpathwaymaybetriggeredbythebindingofmannose-binding
lectin(MBL),ficolin-2orficolin-3todifferentligands.Althoughseveralpapersdemonstratedthesignificanceof
MBLinischemicstroke,theroleofficolinshasnotbeenexamined.
Methods:
Serawereobtainedwithin12hoursaftertheonsetofischemicstroke(admissionsamples)and3-4days
later(follow-upsamples)from65patients.Thecontrolgroupcomprised100healthyindividualsand135patients
withsignificantcarotidstenosis(patientcontrols).Theconcentrationsofficolin-2andficolin-3,initiatormoleculesof
thelectincomplementpathway,weremeasuredbyELISAmethods.ConcentrationofC-reactiveprotein(CRP)was
alsodeterminedbyaparticle-enhancedimmunturbidimetricassay.
Results:
Concentrationsofbothficolin-2andficolin-3weresignificantly(p<0.001)decreasedinboththe
admissionandinthefollow-upsamplesofpatientswithdefiniteischemicstrokeascomparedtohealthysubjects.
Concentrationsofficolin-2andficolin-3wereevenhigherinpatientcontrolsthaninhealthysubjects,indicating
thatthedecreasedlevelsinseraduringtheacutephaseofstrokearerelatedtotheacuteischemicevent.Ficolin-3
levelsinthefollow-upsamplesinverselycorrelatedwiththeseverityofstrokeindicatedbyNIHscaleonadmission.
Infollow-upsamplesaninversecorrelationwasobservedbetweenficolin-3levelsandconcentrationofS100
b
,an
indicatorofthesizeofcerebralinfarct.Patientswithlowficolin-3levelsandhighCRPlevelsinthefollowup
sampleshadasignificantlyworseoutcome(adjustedORs5.6and3.9,respectively)asmeasuredbythemodified
Rankinscalecomparedtopatientswithhigherficolin-3andlowerCRPconcentrations.HighCRPconcentrations
weresimilarlypredictiveforworseoutcome,andtheeffectsoflowficolin-3andhighCRPwereindependent.
Conclusions:
Ourfindingsindicatethatficolin-mediatedlectinpathwaysofcomplementactivationcontributeto
thepathogenesisofischemicstrokeandmaybeadditivetocomplement-independentinflammatoryprocesses.
Keywords:
stroke,ischemicstroke,outcome,complement,lectinpathway,ficolins,ficolin-2,ficolin-3,CRP

Background
injuryinischemicstroke[2-4].Amongotherneuroin-
Neuroinflammationisakeyelementintheischemicflammatoryprocesses,thecomplementsystemisalso
cascadeaftercerebralischemiathatresultsincellactivatedduringtissueinjuryandhasrecentlybeencon-
damageanddeathinthesubacutephase.[1]sideredasanewpotentialtherapeutictargetinischemic
Complementactivationisoneofthepathologicalstroke[5]andinintracerebralhaemorrhage[6].Both
mechanismsthatcontributetotheischemic/reperfusionanimalexperimentsandobservationsmadeinstroke
patientsindicatethatactivationofthecomplementsys-
*Correspondence:[e-mail]
temisoneofthemechanismscontributingtotheexten-
1
3rdDepartmentofInternalMedicine,SemmelwiesUniversity,Budapest,
sionofthecerebralinfarctafterischemicstroke[7].
Hungary
Fulllistofauthorinformationisavailableattheendofthearticle
Severalstudieshavedemonstratedtheessentialroleof
©2011Füstetal;licenseeBioMedCentralLtd.ThisisanOpenAccessarticledistributedunderthetermsoftheCreativeCommons
AttributionLicense(http://creativecommons.org/licenses/by/2.0),whichpermitsunrestricteduse,distribution,andreproductionin
anymedium,providedtheoriginalworkisproperlycited.

Füst
etal
.
JournalofNeuroinfla
m
ation
2011,
8
:185
http://www.jneuroinflammation.com/content/8/1/185

complementactivationinbraindamagefollowingcere-
bralischemia.Suchevidenceincludes(i)anincreased
expressionofcomplementproteinsandcomplement
receptorsafterpermanentmiddlecerebralarteryocclu-
sion(MCAO)[8-11](ii)differentpathologicaleventsin
complement-deficient/-sufficientanimalsaftertheonset
ofcerebralischemiacomparedtowild-typelittermates:
complementdeficientanimalsareatleastpartiallypro-
tectedaftertransientMCAO[12-15].(iii)Inrodent
experimentalmodels,complementdepletioninduced
usingthecobravenomfactor(CVF)[16,17],aswellas
complementinhibitionbyaplasma-derivedC1-inhibitor
[18,19],arecombinantC1inhibitor[20],CR2-Crry[13]
andintravenousimmunoglobulinadministration[14]
wereproventoexertbeneficial,neuroprotectiveeffects,
indicatingtheprotectiveroleofcomplementantagonism
andinhibition.
Onlyafewstudieshaveexploredcomplementactiva-
tioninpatientswithischemicstroke[21,22].Recently,
wefoundthatsC5b-9levelsdeterminedatadmission
exhibitedasignificantpositivecorrelationwiththe
clinicalseverityofstroke,aswellaswiththeextentof
theneurologicaldeficitasdeterminedbydifferent
scales[3].Ourfindingssuggestedthatthelectinpath-
wayisprimarilyresponsiblefortheactivationofcom-
plementinischemicstroke.Inagreementwiththese
findings,Cerveraetal.[4]demonstratedbothinmice
andstrokepatientsthatgeneticallydeterminedMBL-
deficiencyisassociatedwithabetteroutcomeafter
acuteischemicstroke.Inahighnumberofpatients
withischemicstroke,Osthoffetal.[23]foundthata
deficiencyofthemannose-bindinglectinisassociated
withsmallerinfarctionsizeandamorefavorableout-
come.Morerecently,thegroupofDeSimoni[24]
reportedontheformationoffunctionalMBL/MASP-2
complexesinplasmainmiceafterMCAO,and
demonstratedthatmolecules,whichstronglyboundto
MBL,inducedsignificantreductioninneurological
deficitsandinfarctvolume,whenadministered6h
aftertransientMCAO.Thesedatasupportthenotion
thatthelectinpathwayplaysacrucialroleinthe
developmentofischemicstroke.
ApartfromMBL,theficolinsalsoserveasrecognition
moleculesinthelectincomplementpathway.Threedif-
ferentficolinshavebeendescribedinhumans.Ficolin-1,
-2,and-3arederivedfromthegenes
FCN1
,
FCN2
,and
FCN3
,respectively.Inhealthyindividuals,ficolin-2and
-3arepresentintheserumandplasmainrelatively
highconcentrations,whiletheconcentrationofficolin-1
ismuchlower[25].SimilartoMBL,theficolinsare
associatedwithasetofthreeserineproteases,termed
MBL-associatedserineproteases(MASPs),enablingacti-
vationofthecomplementsystem.Theprimaryactivator
ofthelectinpathwayappearstobeMASP-2.

Page2of10

Asdescribedabove,thereareabundantdataaboutthe
significanceofMBLinischemicstroke.Theroleofthe
ficolins,initiatormoleculesofthelectincomplement
pathway,however,hasneverbeenstudiedinthisdis-
ease.Therefore,wemeasuredthelevelsofficolin-2and
ficolin-3inserafrom65patientswithischemicstroke
andfromcontrols.Inordertoassesstheclinicalsignifi-
canceoftheresults,serumconcentrationsofthesepro-
teinswerecorrelatedtoanindirectmeasureofthe
strokeseverity(NIHss),S100
b
concentrationonday3,
whichisanindicatorofthesizeofcerebralinfarct,
[26,27]aswellastheoutcomeofthediseaseexpressed
bythemodifiedRankinscale.
Besidescomplementactivation,otherinflammatory
processesarealsoknowntocontributetothepathogen-
esisoftheischemicstroke[1].Amongthem,CRP-asso-
ciatedprocessesweremostlystudied.In2005,DiNapoli
etal[28]summarizedevidenceforCRPasanindepen-
dentpredictorofcerebrovasculareventsinat-riskindi-
vidualsanditsusefulnessinevaluatingprognosisafter
stroke.ItwasalsodemonstratedthatC-reactiveprotein
predictstheprognosisofpatientswithfunctionaldis-
abilityafterthefirstoccurrenceofischemicstroke[29]
andcorrelatestotheinfarctvolume[30].Recently,
Ormstadetal.[31]providedevidencethatCRPplaysan
importantroleintheprogressionofcerebraltissue
injury.Inaddition,inourpreviousstudy[3]wefound
thatcomplementactivationandelevatedCRPlevels
wereindependentlyassociatedwiththeclinicalseverity
anddifferentoutcomemeasuresofischemicstroke,
indicatingtheiradditiveeffect.Therefore,serumcon-
centrationsofCRPanditsrelationshiptotheficolin
levelswerealsoexaminedhere.
Methods
Patientsandcontrolsubjects
Patientswithischemicstroke
includedinthepresent
workwereadmittedtotwocenters:theDepartmentof
Neurology,Universityof
Pecs
,Hungary(39patients:20
menand19women,aged49-84years)andtheDepart-
mentofNeurology,KútvölgyiClinicalCentre,Semmel-
weisUniversity,
Budapest
,Hungary(26patients:10men
and16women,aged58-87years)
(
Table1).Theman-
agementofischemicstrokewasinaccordancewiththe
guidelinesoftheStrokeCounciloftheAmericanHeart
Association/AmericanStrokeAssociation[32]Noneof
thepatientsweretreatedbyintravenousthrombolysis.
Patientswithstrokewereenrolleduponthefirstoccur-
renceofacuteischemicstrokeonly;allpatientshad
neuroimaging(mostofthembrainMRI,butatleastcra-
nialCT).Nopatientshadhemorrhagicinfarction.All
patientswithdefiniteacuteclinicalsymptomswere
enrolledregardlessofetiologyi.e.lacunarorterritorial
infarctcausedbythrombosisoremboli.Exclusion

Füst
etal
.
JournalofNeuroinfla
m