Cytokine Therapy With Interleukin-2/Anti–Interleukin-2 Monoclonal Antibody Complexes Expands CD4+CD25+Foxp3+ Regulatory T Cells and Attenuates Development and Progression of Atherosclerosis

Background— CD4+CD25+Foxp3+ regulatory T cells (Tregs) attenuate atherosclerosis, but their therapeutic application by adoptive transfer is limited by the need for their expansion in vitro and limited purity. Recently, an interleukin (IL)-2/anti–IL-2 neutralizing monoclonal antibody (IL-2/anti–IL-2 mAb) complex has been shown to expand these Tregs. We examined the capacity of a modified IL-2/anti–IL-2 mAb treatment to expand Tregs and inhibit both the progression and development of developed atherosclerosis. Methods and Results— Six-week old apolipoprotein E–deficient mice fed a high-fat diet for 8 weeks were administered IL-2/anti–IL-2 mAb commencing 2 weeks after starting the diet. Tregs in the spleen, lymph node, and liver were selectively expanded without affecting CD4+, CD8+, or natural killer cells. Tregs were increased in lesions and lesion size reduced. CD4+ T-cells, macrophages, mature dendritic cells, proliferating cell nuclear antigen+ cells, and monocyte chemoattractant protein-1 and vascular cell adhesion molecule-1 were reduced. In anti-CD3–stimulated splenocytes, proliferation and secretion of Th1, Th2, and Th17 (IL-17) cytokines and IL-1&bgr; were reduced. To determine whether treatment attenuated progression of developed atherosclerosis, 6-week-old apolipoprotein E–deficient mice were fed a high-fat diet for 6 weeks, followed by IL-2/anti–IL-2 mAb treatment for 6 weeks while continuing the high-fat diet. Treatment also increased Tregs without affecting CD4+, CD8+, or natural killer cells, suppressed inflammation, and greatly attenuated progression of atherosclerosis. Conclusions— IL-2/anti–IL-2 mAb treatment in vivo attenuates atherosclerosis via selective Tregs expansion. The findings suggest that cytokine-based IL-2/anti–IL-2 mAb complex therapy could represent an attractive approach for treating atherosclerosis, because it markedly attenuates progression as well as development, by modulating its immunoinflammatory component.

[1]  Xiong Ma,et al.  The Interaction between Regulatory T Cells and NKT Cells in the Liver: A CD1d Bridge Links Innate and Adaptive Immunity , 2011, PloS one.

[2]  Steffen Jung,et al.  CCL17-expressing dendritic cells drive atherosclerosis by restraining regulatory T cell homeostasis in mice. , 2011, The Journal of clinical investigation.

[3]  C. Uyttenhove,et al.  B cell depletion reduces the development of atherosclerosis in mice , 2010, The Journal of experimental medicine.

[4]  Da-zhu Li,et al.  The role of CD4+CD25+ regulatory T cells in macrophage-derived foam-cell formation , 2010, Journal of Lipid Research.

[5]  R. Klingenberg,et al.  Intranasal Immunization With an Apolipoprotein B-100 Fusion Protein Induces Antigen-Specific Regulatory T Cells and Reduces Atherosclerosis , 2010, Arteriosclerosis, thrombosis, and vascular biology.

[6]  J. Sprent,et al.  IL-2/anti-IL-2 antibody complexes show strong biological activity by avoiding interaction with IL-2 receptor α subunit CD25 , 2010, Proceedings of the National Academy of Sciences.

[7]  K. Hirata,et al.  Oral Anti-CD3 Antibody Treatment Induces Regulatory T Cells and Inhibits the Development of Atherosclerosis in Mice , 2009, Circulation.

[8]  T. Whiteside,et al.  Generation and Accumulation of Immunosuppressive Adenosine by Human CD4+CD25highFOXP3+ Regulatory T Cells* , 2009, The Journal of Biological Chemistry.

[9]  T. Mrkvan,et al.  In Vivo Expansion of Activated Naive CD8+ T Cells and NK Cells Driven by Complexes of IL-2 and Anti-IL-2 Monoclonal Antibody As Novel Approach of Cancer Immunotherapy1 , 2009, The Journal of Immunology.

[10]  M. Kurrer,et al.  Secretory phospholipase A2-IID is an effector molecule of CD4+CD25+ regulatory T cells , 2009, Proceedings of the National Academy of Sciences.

[11]  E. Shevach.  Mechanisms of foxp3+ T regulatory cell-mediated suppression. , 2009, Immunity.

[12]  James L Riley,et al.  Human T regulatory cell therapy: take a billion or so and call me in the morning. , 2009, Immunity.

[13]  G. Besra,et al.  NKT Cell Subsets Mediate Differential Proatherogenic Effects in ApoE−/− Mice , 2009, Arteriosclerosis, thrombosis, and vascular biology.

[14]  J. Sprent,et al.  In vivo expansion of T reg cells with IL-2–mAb complexes: induction of resistance to EAE and long-term acceptance of islet allografts without immunosuppression , 2009, The Journal of experimental medicine.

[15]  J. Pober,et al.  Interleukin-17 and Interferon-γ Are Produced Concomitantly by Human Coronary Artery–Infiltrating T Cells and Act Synergistically on Vascular Smooth Muscle Cells , 2009, Circulation.

[16]  P. Libby,et al.  The multifaceted contributions of leukocyte subsets to atherosclerosis: lessons from mouse models , 2008, Nature Reviews Immunology.

[17]  Yong‐jun Liu,et al.  Two functional subsets of FOXP3+ regulatory T cells in human thymus and periphery. , 2008, Immunity.

[18]  J. Kirkwood,et al.  Expression of ICOS on Human Melanoma-Infiltrating CD4+CD25highFoxp3+ T Regulatory Cells: Implications and Impact on Tumor-Mediated Immune Suppression1 , 2008, The Journal of Immunology.

[19]  S. Ishihara,et al.  CD4+CD25+Foxp3+ regulatory T cells induce cytokine deprivation–mediated apoptosis of effector CD4+ T cells , 2007, Nature Immunology.

[20]  P. Teeling,et al.  Low Numbers of FOXP3 Positive Regulatory T Cells Are Present in all Developmental Stages of Human Atherosclerotic Lesions , 2007, PloS one.

[21]  A. Steinkasserer,et al.  CD83 Knockdown in Monocyte-Derived Dendritic Cells by Small Interfering RNA Leads to a Diminished T Cell Stimulation1 , 2007, The Journal of Immunology.

[22]  K. Venken,et al.  A CFSE based assay for measuring CD4+CD25+ regulatory T cell mediated suppression of auto-antigen specific and polyclonal T cell responses. , 2007, Journal of immunological methods.

[23]  S. Kaveri,et al.  Human Dendritic Cells Acquire a Semimature Phenotype and Lymph Node Homing Potential through Interaction with CD4+CD25+ Regulatory T Cells1 , 2007, The Journal of Immunology.

[24]  W. van Eden,et al.  Induction of Oral Tolerance to Oxidized Low-Density Lipoprotein Ameliorates Atherosclerosis , 2006, Circulation.

[25]  Da-zhu Li,et al.  Generation of HSP60-specific regulatory T cell and effect on atherosclerosis. , 2006, Cellular immunology.

[26]  B. Toh,et al.  GM-CSF Deficiency Reduces Macrophage PPAR-&ggr; Expression and Aggravates Atherosclerosis in ApoE-Deficient Mice , 2006, Arteriosclerosis, thrombosis, and vascular biology.

[27]  J. Sprent,et al.  Selective Stimulation of T Cell Subsets with Antibody-Cytokine Immune Complexes , 2006, Science.

[28]  R. Flavell,et al.  Natural regulatory T cells control the development of atherosclerosis in mice , 2006, Nature Medicine.

[29]  A. Scheffold,et al.  Regulation of CD4+CD25+ regulatory T cell activity: it takes (IL‐)two to tango , 2005, European journal of immunology.

[30]  Shimon Sakaguchi,et al.  Homeostatic maintenance of natural Foxp3 + CD25+ CD4+ regulatory T cells by interleukin (IL)-2 and induction of autoimmune disease by IL-2 neutralization , 2005, The Journal of experimental medicine.

[31]  M. Rudling,et al.  Lesion Development and Response to Immunization Reveal a Complex Role for CD4 in Atherosclerosis , 2005, Circulation research.

[32]  Matthias Edinger,et al.  Only the CD62L+ subpopulation of CD4+CD25+ regulatory T cells protects from lethal acute GVHD. , 2005, Blood.

[33]  S. Gaffen,et al.  Overview of interleukin-2 function, production and clinical applications. , 2004, Cytokine.

[34]  C. Drake,et al.  Role of LAG-3 in regulatory T cells. , 2004, Immunity.

[35]  P. Tipping,et al.  The role of interleukin-4 and interleukin-12 in the progression of atherosclerosis in apolipoprotein E-deficient mice. , 2003, The American journal of pathology.

[36]  A. Rudensky,et al.  Foxp3 programs the development and function of CD4+CD25+ regulatory T cells , 2003, Nature Immunology.

[37]  C. Come,et al.  Influence of Interferon-&ggr; on the Extent and Phenotype of Diet-Induced Atherosclerosis in the LDLR-Deficient Mouse , 2003, Arteriosclerosis, thrombosis, and vascular biology.

[38]  F. Liew,et al.  CD4+CD25+ Regulatory T Cells Suppress Differentiation and Functions of Th1 and Th2 Cells, Leishmania major Infection, and Colitis in Mice1 , 2003, The Journal of Immunology.

[39]  N. Kalinina,et al.  Cytochrome b558—Dependent NAD(P)H Oxidase—Phox Units in Smooth Muscle and Macrophages of Atherosclerotic Lesions , 2002, Arteriosclerosis, thrombosis, and vascular biology.

[40]  W. Strober,et al.  Cell Contact–Dependent Immunosuppression by Cd4+Cd25+Regulatory T Cells Is Mediated by Cell Surface–Bound Transforming Growth Factor β , 2001, The Journal of experimental medicine.

[41]  E. Shevach,et al.  Cutting Edge: Control of CD8+ T Cell Activation by CD4+CD25+ Immunoregulatory Cells , 2001, The Journal of Immunology.

[42]  Xinghua Zhou,et al.  Transfer of CD4+ T Cells Aggravates Atherosclerosis in Immunodeficient Apolipoprotein E Knockout Mice , 2000, Circulation.

[43]  A. Becker,et al.  Cytokine secretion profiles of cloned T cells from human aortic atherosclerotic plaques , 1999, The Journal of pathology.

[44]  B. Rollins,et al.  MCP-1 deficiency reduces susceptibility to atherosclerosis in mice that overexpress human apolipoprotein B. , 1999, The Journal of clinical investigation.

[45]  G. Hansson,et al.  Hypercholesterolemia is associated with a T helper (Th) 1/Th2 switch of the autoimmune response in atherosclerotic apo E-knockout mice. , 1998, The Journal of clinical investigation.

[46]  G. Hansson,et al.  Evidence for a local immune response in atherosclerosis. CD4+ T cells infiltrate lesions of apolipoprotein-E-deficient mice. , 1996, The American journal of pathology.

[47]  F. Finkelman,et al.  Anti-cytokine antibodies as carrier proteins. Prolongation of in vivo effects of exogenous cytokines by injection of cytokine-anti-cytokine antibody complexes. , 1993, Journal of immunology.

[48]  R. Williams,et al.  Quantitative assessment of atherosclerotic lesions in mice. , 1987, Atherosclerosis.

[49]  C. McSharry,et al.  CD4 CD25 Regulatory T Cells Suppress Differentiation and Functions of Th1 and Th2 Cells, Leishmania major Infection, and Colitis in Mice , 2002 .