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Homostatische,Mechanismen,der,,CD4+,T6Zellgenese,bei,Rheumatoider,Arthritis,

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Dissertation,zur,Erlangung,des,akademischen,Grades,

Dr.,med.,,

an,der,Medizinischen,Fakultt,,der,Universitt,Leipzig,

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!!eingereicht!von:!Annika!Schatz!geboren!am!08.10.1986!in!Bad!Kreuznach!!angefertigt!an!der!Medizinischen!Klinik!II!der!Universitt!Leipzig!!Klinik!und!Poliklinik!fr!Gastroenterologie!und!Rheumatologie!!Sektion!Rheumatologie!!!

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Betreuer:!Prof.!Dr.!med.!Ulf!Wagner!und!Dr.!rer.!nat.!Manuela!Rossol!!

Beschluss!ber!die!Verleihung!des!Doktorgrades!vom:!22.11.2016!

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!

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!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!Gewidmet!!

!

meinem!Grovater!Dietrich!Bunner,!!

meinem!Vater!Dieter!Schatz!!

und!!

meinem!Mann!Patrick!Jahnke!

Bibliographische.Beschreibung..Schatz,(Annika(Katrin(

(

Homostatische.Mechanismen.der.CD4+.T

Inhaltsverzeichnis.

.

Abkrzungsverzeichnis..................................................................................................I((

Tabellen................................................................................................................................II((

Abbildungen......................................................................................................................III(

1.(Einleitung.......................................................................................................................1(

((1.1.(Epidemiologie,(tiologie(und(Pathogenese(der(Rheumatoiden(Arthritis........................1(

((1.2.(TLZellen(in(der(Pathogenese(der(Rheumatoiden(Arthritis...............................................2(

((1.3.(Die(TLZellgenese...............................................................................................................4(

(((((1.3.1(Entwicklung(im(Knochenmark:(von(der(hmatopoetischen(Stammzelle(zum(((

((((((((((((((common(lymphoid(progenitor((CLP)............................................................................4(

(((((1.3.2(Lymphoide(Vorluferzellen(in(der(peripheren(Zirkulation..........................................6(

(((((1.3.3(Die(Entwicklung(von(CD4+(und(CD8+(einfach(positiven(TLZellen(im(Thymus..............7(

((1.4.(Immunoseneszenz(und(das(periphere(CD4+(TLZellkompartiment(bei(Rheumatoider((

(((((((((Arthritis..........................................................................................................................12(

(

2.(Material(und(Methoden.......................................................................................17(

((2.1.(Patientengut(und(gesunde(Kontrollen...........................................................................17(

((2.2.1.(Gewinnung(der(PBMCs...............................................................................................17(

((2.2.2.(Gewinnung(der(PBMCs(fr(das(CD31LExperiment......................................................19(

((2.3.(Immunzytologische(Frbung..........................................................................................20(

((2.4.(Etablierung(der(Frbeanstze........................................................................................21((

((2.5.(Messung(am(Durchflusszytometer................................................................................22(

((2.6.(Auswertung(der(Messergebnisse(der(Durchflusszytometrie.........................................24(

(((((2.6.1(Messungen(1(bis(4.....................................................................................................24(

(((((2.6.2.(Messung(5................................................................................................................25(

(((((2.6.3.((Messung(6...............................................................................................................26(

2.7.(Erfassung(der(klinischen(Daten(der(Patienten.................................................................28(

2.8.(Statistische(Auswertung(der(Ergebnisse..........................................................................30(

(

3.(Ergebnisse...................................................................................................................31(

((3.1.(Beschreibung(des(CD4+(TLZellkompartiments(in(der(peripheren(Zirkulation................31(

(((((3.1.1.(CD4+CD8+(doppelt(positive(TLLymphozyten(in(der(peripheren(Zirkulation............31(

(((((3.1.2.(Frequenz(CD4+(CD28(negativer(TLLymphozyten(in(der(peripheren(Zirkulation.......33(

(((((3.1.3.(Untersuchung(der(CD85j(positiven(TLHelferzellpopulation.....................................35(

(((((3.1.4.(CD4+CD25+(TLZellen:(aktivierte(TLHelferzellen(und(TLregulatorische(Zellen...........38(

(((((3.1.5.(Quantifizierung(von(naiven(versus(MemoryLTLHelferzellen....................................40(

((3.2.(Recent(thymic(emigrants:(CD31+(thymic(naive(TLZellen...............................................42(

((3.3.(Zirkulierende(lymphoide(Progenitoren..........................................................................44(

((3.4.(CD34(positive(hmatopoetische(Stammzelle................................................................48(

(

4.(Diskussion....................................................................................................................52(

4.1.(Das(periphere(CD4+(TLZellkompartiment(bei(der(RA......................................................52(

4.2.(RALPatienten(weisen(eine(verringerte(Anzahl(von(RTEs(auf...........................................67(

4.3.(Die(Frequenz(von(Prthymozyten(im(peripheren(Blut(von(RALPatienten(ist(erhht.......70(

(

5.(Zusammenfassung..................................................................................................76(

(

Literaturverzeichnis.....................................................................................................79(

(

Eigenstndigkeitserklrung......................................................................................89(

(

(

Danksagung......................................................................................................................90(

(

(

.

.

.

(Abkrzungsverzeichnis.................................................................................I.

(

Abb.(AIRE((APC(ATM(bzw.(C(CCP(CCR(CD(CLP(CMP(CMV(CRP(CTL(CXCR(DAS(DMARD(DN(DP(EBV(EDTA(ELP(et(al.(EULAR(FITC(FKS(Flt3(FoxP3(g(GPCR(

Abbildung(autoimmune(regulator(Allophycocyanin,(antigen(presenting(cell((Ataxia(telangiectasia(mutated(beziehungsweise(Celcius(cyklisches(citrulliniertes(Peptid(CCLMotivLChemokinLRezeptor(cluster(of(differentiation(common(lymphoid(progenitor(common(myeloid(progenitor(Cytomegalievirus(CLreaktives(Protein(cytotoxic(T(lymphocytes(CXCLMotivLChemokinrezeptor(disease(activity(score(Disease(modifying(antiLrheumatic(Drugs(doppelt(negativ(doppelt(positiv(EpsteinLBarrLVirus(Ethylendiamintetraessigsure(early(lymphoid(progenitor(et(alii(European(league(against(rheumatism(fluorescein(isothiocyanate(fetales(Klberserum(fmsLlike(tyrosine(kinase(3(forkhead(box(P3(Gramm(GLProtein(gekoppelter(Rezeptor(

HLA((HSC(ICAM(IDO(IFN(Ig(IL(IL7R(ILTL2(IPEX((ITAM(ITIM(kDA(KIR(Lck(LILRB1(LIR1(LMPP(LTLHSC(LYP(MCP(MHC(ml(mM(mm(MPP(mRNA(l(NKG2D(NKLZelle(PBMC(PBS(PE(PECAM(PIP(PSGL(PT(RA(RAG(RT(RTE(Runx(S(SEM(SFK(SP(

Humanes(Leukozytenantigen(hmatopoetische(Stammzelle(Intercellular(Adhesion(Molecule(IndolaminL2,3LDioxygenase(Interferon(Immunoglobulin(Interleukin(InterleukinL7LRezeptor((IgLlike(transcript(2(immunodysregulation(polyendocrinopathy(enteropathy(XLlinked(syndrome(immunoreceptor(tyrosineLbased(activation(motif(immunoreceptor(tyrosineLbased(inhibition(motif(Kilodalton(KillerLcell(immunoglobulinLlike(receptor(lymphocyteLspecific(protein(tyrosine(kinase(leukocyte(immunoglobulinLlike(receptor(1,(subfamily(B(leukocyte(immunoglobulinLlike(receptor(1(lymphoidLprimed(multipotent(progenitor(longLterm(haematopoietic(stem(cell(lymphoide(Phosphatase(Metacarpophalangealgelenk(major(histocompatibility(complex(Milliliter(millimolar(Millimeter(multipotent(progenitor(messengerRibonukleinsure(Mikroliter(naturalLkiller(group(2,(member(D(Natrliche(Killerzelle(peripheral(blood(mononuclear(cell(PhosphateLbuffered(saline(Phycoerythrin(Platelet(endothelial(cell(adhesion(molecule(proximales(Interphalangealgelenk(PLSelectin(Glykoproteinligand(Prthymozyt(Rheumatoide(Arthritis(Recombination(activating(gene(Raumtemperatur(recent(thymic(emigrant(RuntLrelated(transcription(factor(Seite(standard(error(of(the(mean(Src(family(kinase(single(positive((

STLHSC(SW(TCR(TdT(TEN(ThPOK(TNF(TREC(Treg(u.a.(VAS(VCAM(vs(z.B.(

shortLterm(heamatopoietic(stem(cell(swollen(joint(TLZellrezeptor(terminale(Desoxnukleotidytransferase(tender(joint(Th(inducing(POZLKruppel(Factor(Tumornekrosefaktor((TLcell(receptor(excision(circle(regulatorische(TLZelle(und(andere(visuelle(Analogskala(vascular(cell(adhesion(molecule(versus(zum(Beispiel(((((((

Tabellen..........................................................................................................II.

(

Abbildungen.................................................................................................III.

.

Tabelle(1( Hinweise(auf(die(entscheidende(Rolle(der(CD4+(TLZellen(fr(die(Pathogenese(der(Rheumatoiden(Arthritis((

( S.(3(

Tabelle(2( Einteilung(der(MPPs(anhan