11/7/2011. Disclosures. Psoriatic Arthritis (PsA) DC-STAMP I II III IV. None
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1 unstimulte stimulte 11/7/11 Ientifiction of Unique Suset + (Denritic Cell-Specific Trnsmemrne Protein) T cells with Th17 Signture in Psoritic rthritis () Ptients Disclosures None Y.H. Chiu, E.M. Schwrz, W. Wojciechowski,. Pnepento, I. Kuzin, T. ushnell, S. Moorehe, R. rrett, n C.T. Ritchlin Psoritic rthritis () -- inflmmtory joint isese Increse frequency of circulting osteoclst precursors (OCP) in ptients HC -- one erosions re common -- osteoclsts meite one resorption Ritchlin, CT, et l., 3, J. Clin. Invest, 111(6): Four mjor ptterns in humn PMC I II III IV * Require for fusion of OCPs to form OC * Expresse on the surfce of * OCP mrker * ITIM is present on cytoplsmic til (signling) Chiu, Y.H., et l., Journl of one n Minerl Reserch, online Octoer, 11 (N=1) HC (N=) OC# Chiu, Y.H., et l., Journl of one n Minerl Reserch, online Octoer, 11 1
2 CD3 <Violet C 4/->: CD3-P + 11/7/11 lthough is minly expresse on, smll suset of T cells lso express CD14+ CD3+ CD <lue F /->: DCSTMP-FITC live Specimen_1_D PMC.fcs Chiu, Y.H., et l., Journl of one n Minerl Reserch, Online Octoer, Th17 in Psorisis (Ps) n Psoritic rthritis () 1) Th17 cells promote inflmmtion, crtilge egrtion n osteoclstogenesis (Sto., et l., 6). ) Th17 cells infiltrte psoritic skin n synovium (Rychuhuri, et l., 11; Zl. C., et l., 9). 3) Th17 cells re elevte in the circultion n synovil flui of ptients (Leipe et l., 1). 4) gents tht inhiit the IL-3/Th17 re effective in n Ps (Goron, K.., et l., 11; Solt, L, et l., 11; Gottlie, 9). Dr. uckner s tlk n strct # 779 Rtionle: Given tht 1) Th17 cells re increse in ) expression is enhnce in suset of T cells Hypothesis: ++ cells elong to the Th17 linege. + + cells re expne in ptients Results Helthy Control (HC) 87% 13% % %.1
3 Frequency CD3 % of + + cells in PMC % of +DCSTMP+ in PMC reltive gene expression (normlize to ctin) IL-17 (pg/ml) <Green 71/->: -PECy CCR4 CCR4 CCR4 <Green 78/4->: CCR4-PECy CD <Violet G /4->: IL3R-io-PO <Violet G /4->: IL3R-io-PO <Green 78/4->: CCR4-PECy <Re C 66/->: CD161-PC <Re C 66/->: CD161-PC CD <Violet H 4/->: IL17-P <Violet H 4/->: IL17-P 11/7/ cells re elevte in HC. ± 3.1 (n=4) * HC PsC PsC 11. ± 7. (n=19) * 1.3 ± 8.1 (n=11) * < () <lue 1/->: DCSTMP-FITC + + cells express Th17-specific mrkers () + + () - () () IL-3R CCR6 CD161,31 7,676 () IL-17 1,11 3, cells re preferentilly expne in Th17-promoting conitions plin IL- Th The Th17 signture of + + cells is enhnce y Th17-promoting culture conitions Fresh PMC 11% (i) +- (ii) ++ IL-3R surfce expression ROR-gt y qrt-pcr IL17 ELIS fter -y in Th-17 promoting mei CCR6 3 RORgt IL-3R plin +IL- Th mei IL Th17 7% CCR6 Imge strem nlysis confirms istinct cell size n locliztion in CD14+ n CD3+ cells gte on live cells (FITC) CD14 (PE) (PerCy.) Summry 7-D expression: n suset of T cells ++ T cells mnifest Th17 signture CD14 n Th17 specific mrkers re up-reregulte y Th17-promoting culture conitions T cells + + T cells re significntly elevte in the circultion of ptients 3
4 % of ++ cells 11/7/11 Conclusion IR Division:. Pnepento I. Kuzin S. Moorehe R. rrett J. Hosseler CTR MSK Reserch: Eie M. Schwrz Ywen Ju T.J. Hsu Flow Core: W. Wojciechowski T. ushnell + + T cell suset my provie insight into pthogenesis n my e use s isese iomrker Sponsors: NIH NPF IPRT chnge in ++ % fter nti-tnf tretment osteolst Unknown lign Immune complex IFNg LPS TNF ctivte RNKL RNK CD TRF6 NFTc1 cfos P ITIM Shp1 Shc Dok? ITM P Syk Sos Rs efore nti-tnf gent 1 fter nti-tnf gent cell fusion OC ifferentition one resorption C+ signl PLCg numers: DSI score Figure 7 Person correltion=.6, p=.3, n=14 4
5 counts CD14 <Re C 66/->: CD14-PC 7-D <lue 67LP->: 7D FSC <lue E 3/3->: DCTSMP-FITC 8. <Re C 66/->: CD14-PC SSC FSC <lue E 3/3->: DCTSMP-FITC <Re C 66/->: CD14-PC CD <lue E 3/3->: DCTSMP-FITC <Violet C 4/->: CD3-P <lue E 3/3->: DCTSMP-FITC counts OC count (per 1 6 ) 11/7/11 1 unstine isotype C lnce of Th17 n Treg: 8 6 CD14+CD16- CD14+CD16+ Th17 n Treg is lnce in psoritic sites KD <lue F /->: DCSTMP FITC D : without 1 : with 1 c c 489±84 * p=.13 * * 6± in the culture Figure FSC c (P1) P1 (P) P FSC CD CD3+D 6 CD19+D P3 e (P3) isotype control P1 P P3 f Figure pttern # of sujects helthy control ptients OC# (per 1 6 PMC) I II III IV I II III IV mein inter-qurtile rnge Figure 3 Fresh Dy 1 Dy Dy Dy 7 c e enriche lymphocytes enriche C 16 hrs 3 ys no DN vector lone + c e f Figure 4 C Figure Low lymphocytes FSC High FITC- 1 1x1 D low () high (16) c
6 cell counts <lue D 8/4->: PLCg-PE <lue D 8/4->: PLCg-PE <lue D 8/4->: PLCg-PE /7/11 () I y nti- ntioy : mrker : IP y 1 3: IP y nti-cd16 () I y nti- ntioy (C) I y nti-phosphotyrosine ntioy 4G : mrker : IP y nti-shp1 3: IP y mouse IgG (D) Effect of 1 tretment on signling ( I ) 1 ( II ) hrs 4 hrs 64 hrs 1 e 46% 1: mrker : cells without 1 3: cells trete with 1 4. cells c without 1 : cells trete 4% 3% 77% with 1 49% 1% 6% 4% 19% 81% f IP y IP y mouse nti-sh mei lone : mrker : IP y IgG 3: IP y nti-shp-1 4. IP y 1 g h 4% 46% + IgG nti-phosphorylte PLC-g i 6% 74% Figure 6 6
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% ells with ili (mrke y A-Tu) Reltive Luiferse % ells with ili (mrke y Arl13) % ells with ili DOI: 1.138/n2259 A-Tuulin Hoehst % Cilite Non-ilite -Serum 9% 8% 7% 1 6% % 4% +Serum 1 3% 2% 1% % Serum: -
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2 weeks high holesterol diet 2 weeks high holesterol diet 2 weeks high holesterol diet 2 μm Mrophges Crystls Hoehst μm Mrophges Crystls Hoehst Hoehst Crystls Mrophges 2 μm 2 μm Supplementry Fig. 1: Erly
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Supplementry Figure S Connexin4 TroponinI Merge Plsm memrne Met Intrcellulr Met Supplementry Figure S H9c rt crdiomyolsts cell line. () Immunofluorescence of crdic mrkers: Connexin4 (green) nd TroponinI
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Supplementry Figure S1 d MAP2 GFAP e MAP2 GFAP GFAP c f Clindin GFAP Supplementry Figure S1. Neuronl deth nd ltered strocytes in the rin of n ffected child. Neuron specific MAP2 ntiody stining in the hippocmpus
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