Development and functions of patrolling monocytes and kidney resident macrophages

2 Acknowledgements . 3 Declaration . 6 Abbreviations . 13 Table of Figures, Table of Tables. 12 1 Introduction . 17 1.1 Historical perspective . 18 1.1.1 Immunity to disease as a passive phenomenon . 18 1.1.2 The emergence of a cellular immune system . 20 1.1.3 Classification of phagocytes into unified systems : from the ReticuloEndothelial System, to the Mononuclear Phagocyte System . 23 1.1.4. New paradigm and the contribution of studies on fetal hematopoiesis . 27 1.2. Monocyte Subsets and Functions . 28 1.2.1. Human subsets . 28 1.2.2. Mouse subsets . 30 1.2.3. Ly6C monocytes functions . 32 1.2.3.1. In bacterial infections . 32 1.2.3.2. In parasitic infections . 34 1.2.3.3. In fungal infections . 35 1.2.3.4. In viral infections . 36 1.2.3.5. During cancer . 37 1.2.3.6. During sterile inflammation or injury . 38 1.2.4. Ly6C monocyte functions . 40 1.2.4.1. Patrolling activity . 40 1.2.4.2. Ly6C monocytes in infectious diseases and cancer . 43 1.2.4.3. Role of Ly6C monocytes in other immunological disorders . 44 1.2.4.4. Ly6C monocytes as mediators of IgG-dependent functions . 46 1.3. Monocyte development . 47 1.3.1. Hematopoiesis, overview . 47 1.3.2. MDP and cMoP are steps towards differentiated monocytes . 50 1.3.3. Relationship of monocytes with dendritic cells and macrophages . 53 1.3.4. Molecular control of monocyte development and exit from the bone marrow . 55 8 1.3.4.1 Molecular control of monocytes development . 55 · CSF-1/CSF-1R axis . 55 · PU.1 . 57 · Role of the IRF8 / KLF4 cascade in the lineage specification towards monocytes . 58 · NR4A1 controls Ly6C monocyte differentiation . 60 1.3.4.2. Control of monocyte exit from the bone marrow . 62 · CCR2 and the egress of Ly6C monocytes from the bone marrow . 62 · S1PR5 controls the exit of Ly6C monocytes from the bone marrow . 63 1.4 Tissue Macrophages . 64 1.4.1. Macrophage development and maintenance . 64 1.4.1.1. Embryonic Hematopoiesis, overview . 64 1.4.1.2. Development of tissue resident macrophages from an Erythro-Myeloid Progenitor (EMP) . 68 1.4.1.3. Macrophage maintenance throughout adult life . 73 1.4.2. Macrophage functions in various tissues and conditions . 75 1.4.2.1 Macrophage function in homeostasis . 75 1.4.2.2. Macrophage function during various perturbations of steady state . 77 1.4.3. Kidney macrophages in health and disease . 79 1.4.3.1 Tissue localisation and definition of myeloid populations in the kidney 79 1.4.3.2. Kidney macrophage functions in different contexts . 80 1.5. On mathematical modeling . 82 1.5. 1. On the efficiency of mathematics in science . 83 1.5. 2. Modeling Biology with Mathematics . 85 1.6. Preceeding work, general aims, and experimental approach . 87 1.6. 1. Study of monocyte dynamics in the steady state toward anin silico model . 87 1.6. 1.1. Previous results in the laboratory regarding monocytes in the steady state . 87 1.6. 1.2. Aim, and experimental approach . 88 1.6. 2. Study of monocyte and macrophage functions in the kidney . 89 1.6. 2.1. Monocytes as housekeepers of the vasculature, and kidney macrophages as monitors of circulation . 89 1.6. 2.2. Aim and experimental approach . 89 2. Materials and Methods . 91 2.1. Mice . 91 9 2.1.1. Animals used for this study . 91 2.1.2. Genotyping . 92 2.1.2.1. Nr4a1; Nr4a1 . 92 2.1.2.2. Cx3cr1 . 92 2.1.2.3. Rag2 Il2rg . 93 2.1.2.4. Csf1r Fcgr4 . 93 2.2. Flow Cytometry . 94 2.2.1. Tissue preparation for flow cytometry . 94 2.2.1.1. Blood preparation . 94 2.2.1.2. Spleen preparation for monocyte staining . 95 2.2.1.3. Bone marrow preparation . 95 2.2.1.4. Kidney, Liver, Spleen, Lungs, Brain for staining macrophages . 96 2.2.1.5. Skin preparation for epidermal Langerhans cell and dermal macrophages isolation . 97 2.2.2 Flow cytometry acquisition . 97 2.3. EdU incorporation Experiments. 98 2.3.1. EdU administration for single-pusle study . 98 2.3.2. Common protocol for all organs for EdU detection . 98 2.4 Cytospin . 100 2.5. CyTOF . 101 2.6. Absolute cell numbers . 104 2.6. 1. Cell preparation . 104 2.6. 1.1. Blood . 104 2.6. 1.2. Bone Marrow . 104 2.6. 1.3. Spleen . 104 2.6. 2. Cell count using Neubauer Chambers . 105 2.6. 3. Whole body cells numbers per organ .