Immune Cells in Atherosclerosis Regulatory vs Inflammatory T cells Göran K Hansson
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1 Immune Cells in Atherosclerosis Regulatory vs Inflammatory T cells Göran K Hansson Center for Molecular Medicine Karolinska Institute Stockholm, Sweden
2 DECLARATION OF CONFLICT OF INTEREST Goran Hansson has filed patent applications regarding the use of immunotherapy to treat atherosclerosis.
3 T cells and macrophages accumulate at sites of LDL retention in the forming atherosclerotic plaque Libby, Ridker & Hansson, Nature 2011
4 LDL reactive T cells in human plaque Th1 cells that produce inflammatory cytokines - IFNg, TNFa LDL GK Hansson N Engl J Med 2005; 352:
5 Specificity of LDL reactive T cell hybridomas Hermansson, Ketelhuth et al J Exp Med 2010 T cells from LDL immunized mice recognize apob100 of native LDL, but not oxldl!
6 T cell antigen receptors (TCR) recognize antigens of (native, not oxidized) LDL Majority of LDL specific T cells in mice are of TRBV31 type LDL GK Hansson N Engl J Med 2005; 352:
7 Immunization with TRBV31 peptide induces TCR blocking antibodies Anti-TRBV31 impairs T cell response to ApoB100 APC MHC TCR T Hermansson, Ketelhuth et al J Exp Med 2010
8 Immunization against TRBV31 reduces atherosclerosis in hypercholesterolemic mice HuB100tg x Ldlr- mice immunized w/ TRBV31 peptide-klh Western diet 10 wk 57% lesion reduction in aortic arch 65% lesion reduction in aortic root Hermansson, Ketelhuth et al J Exp Med 2010
9 LDL reactive Th1 cells promote atherosclerosis LDL reactive T cells recognize peptide elements of native LDL via TRBV31-type TCR LDL GK Hansson N Engl J Med 2005; 352:
10 Treg cells inhibit proinflammatory immunity Operate via TGF-b, IL-10 LDL GK Hansson N Engl J Med 2005; 352:
11 Role of regulatory T cells in atherosclerosis Cytokines produced by Treg are antiatherosclerotic IL-10, TGF-b Mallat 1999; Caligiuri 2001; Robertson 2003; Gojova 2003 Ldlr -/- mice with reduced Treg (ICOS-; CD28-) develop increased atherosclerosis Gotsman 2006; Ait-Oufella 2006 Transfer of CD4 + CD25 + T cells enriched in Treg reduces disease Ait-Oufella 2006 These findings suggest an important role for Treg
12 The transcription factor FoxP3 drives Treg differentiation S. Sakaguchi, Nat Imm 2005
13 DEREG mice carry diphtheria toxin receptor + GFP under FoxP3 promoter - Selective ablation of Treg by inj diphtheria toxin FoxP3 promoter K Lahl, T Sparwasser et al J Exp Med 2007 CD3 GFP FoxP3 LN cells
14 Treg deficiency by transplantation of DEREG bone marrow into Ldlr -/- mice Donor mice wt DEREG + Treatment Harvest Bone marrow Ldlr -/- Recipient mice (8-10 weeks old) 6 weeks engraftment & recovery Vehicle Hi-fat diet 8 weeks + DT GFP+ Treg no depletion Treg depletion
15 Increased atherosclerosis after Treg elimination in DEREG - Ldlr -/- chimeras Lesion mm 2 / Total wall mm 2 in cross-sections of proximal aorta PBS PBS DT DT Klingenberg, Gerdes et al Subm, 2011
16 Treg cells inhibit atherosclerosis LDL GK Hansson N Engl J Med 2005; 352:
17 Dendritic cells patrol artery, move to lymph nodes, and activate naïve T cells IL-10 Treg cells Th1 effector cells Hansson & Hermansson, Nature Immunol 2011
18 DC therapy Bone marrow from mice Exp. Groups DC (no antigen) Culture with GM- CSF and IL-4 Enrich DC with CD11c MACS separation column Inject pulsed DCs into hub100tg x Ldlr-/- mice (once, I.V.) DC + ApoB100 DC + ApoB100 + IL-10 DC + IL-10 (no antigen) Pulse DC with ApoB100 and IL-10 together with LPS Hermansson et al Circ 2011
19 foxp3 IL-10 foxp3 % of Max Tolerogenic DC induce Treg and inhibit Figure Teffector 2 responses A DC 2.56IL-10 Anti- CD3 control + IL DC IFNg DC + IL B CFSE w/ C Conventional DC CFSE Proliferation of T effector cells w/ ToleroDC D Hermansson et al, Circ 2011
20 Tolerogenic DC reduce atherosclerosis Hermansson et al Circ 2011
21 Induction of Tolerogenic DC oxldl oxldl LDL FoxP3+ Treg Dendritic cell IL-10 TGF-b TRAV_X TRBV31 TRBV31 B Induction of TCR blocking antibodies T X IFNγ X IL-12 TNFα Inflammation and Atherosclerosis
22 Roland Klingenberg Norbert Gerdes Daniel Ketelhuth Andreas Hermansson Daniel Johansson Gabrielle Berne Cardiovascular Research Laboratory Xinghua Center for Molecular Medicine, Karolinska Zhou Institutet
23 Collaborators: Antonino Nicoletti Jan Nilsson Ziad Mallat Alain Tedgui Tim Sparwasser Funding: Vetenskapsrådet Hjärt-Lungfonden Stiftelsen för Strategisk Forskning Vinnova European Union Leducq Foundation Cardiovascular Research Laboratory Center for Molecular Medicine, Karolinska Institutet
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Supplemental Table I Male / Mean ± SEM n Mean ± SEM n Body weight, g 29.2±0.4 17 29.7±0.5 17 Total cholesterol, mg/dl 534.0±30.8 17 561.6±26.1 17 HDL-cholesterol, mg/dl 9.6±0.8 17 10.1±0.7 17 Triglycerides,
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