Long-term innate immune memory via effects on bone marrow progenitors
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1 Long-term innate immune memory via effects on bone marrow progenitors Helen S Goodridge, PhD helen.goodridge@csmc.edu Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, USA Fondation Mérieux Off-Target Effects of Vaccination June 2015
2 Macrophage Origins yolk sac yolk sac-derived tissue-resident macrophages Primitive Hematopoiesis - yolk sac HSC-derived Definitive Hematopoiesis - fetal liver - bone marrow bone marrow HSC tissue-resident macrophages monocyte HSC-derived inflammatory macrophages
3 Inflammatory gene regulation TNF-a Pam 3 CSK 4 time tolerance TNF-a LPS Pam 3 CSK 4 time epigenetic modifications metabolic changes TNF-a training time b-glucan Pam 3 CSK 4
4 Macrophage Memory? Epigenetic and metabolic changes could underlie trained immunity in long-lived macrophages But what about short-lived macrophages? Could changes in macrophage function be maintained via programming of their precursors i.e. hematopoietic stem cells and progenitors? HSC myeloid progenitors monocyte macrophage
5 microbial components IL-3 IL-6 IFN-g G-CSF M-CSF GM-CSF Flt3L Hematopoietic Stem and Progenitor Cells (HSPCs) neutrophils monocytes cdcs and pdcs macrophages modcs
6 Microbial Detection by HSPCs HSPCs express pattern recognition receptors, including Toll-like receptors (TLRs) e.g. HSC CLP CMP GMP TLR2 TLR2 TLR2 Nagai et al. (2006) Immunity 24: TLR2 TLR stimulation can drive or promote production of neutrophils, macrophages, DCs, NK cells in vitro and in vivo in the steady-state and under emergency conditions
7 Does exposure of HSPCs to TLR agonists alter the function of the macrophages they produce? Detection of a TLR2 agonist by hematopoietic stem and progenitor cells impacts the function of the macrophages they produce Yanez et al. (2013) European Journal of Immunology 43:
8 mouse HSPCs = Lin- cells Myelopoiesis
9 Cytokine production by TLR2-derived and M-CSF-derived macrophages mouse Lin - cells (HSPCs) d0-6 M-CSF or d0-6 Pam 3 CSK 4 sort adherent macrophages, and assay cytokines
10 Cytokine production by macrophages derived from HSPCs exposed to a TLR2 agonist in vivo d4 ex vivo LPS stimulation
11 Macrophage production by HSPCs exposed to a TLR2 agonist d0 Pam 3 CSK 4 M-CSF HSPCs d1 wash harvest macrophages and assay
12 Cytokine production by macrophages derived from HSPCs exposed to a TLR2 agonist
13 Cytokine production by macrophages derived from HSPCs exposed to a TLR2 agonist LT-HSC ST-HSC MPP CMP GMP d16 Pam 3 CSK 4 d14 Pam 3 CSK 4 d7 Pam 3 CSK 4
14 Are macrophages derived from TLR2 agonist-exposed HSPCs tolerized? Pam 3 CSK 4 zymosan Pam 3 CSK 4 zymosan mac mac HSPC mac reactive oxygen reactive oxygen
15 Macrophages derived from TLR2 agonistexposed HSPCs do not appear to be immature
16 TNF-a (pg/ml) RLU TLR2 stimulation of human HSPCs programs macrophage function macrophages derived from human CD34 + cells 2000 reactive oxygen ** unstim. Pam 3 CSK 4 LPS d23 stimulus ** d23 zymosan stimulation (minutes) d0 none d0 Pam 3 CSK ng/ml d0 Pam 3 CSK 4 1 mg/ml d0 Pam 3 CSK 4 10 mg/ml d0 none d0 Pam 3 CSK 4
17 TLR2 agonist Progenitors??? Monocytes Macrophages increased macrophage yield reduced cytokine and ROS production upon stimulation
18 IL-6 (ng/ml) IL-6 (ng/ml) TNF-a (ng/ml) TNF-a (ng/ml) Effects of other microbial components on programming of macrophage function * Lin- cells Day 0 treatment Day 7 Pam 3 CSK 4 Day 7 LPS M-CSF M-CSF + Pam 3 CSK M-CSF + LPS ** * Day 7 Pam 3 CSK 4 Day 7 LPS Gil et al. unpublished
19 IL-6 (ng/ml) IL-6 (ng/ml) TNF-a (ng/ml) TNF-a (ng/ml) Effects of other microbial components on programming of macrophage function 4 3 ** * Lin- cells Day 0-7 treatment Day 7 Pam 3 CSK 4 Day 7 LPS M-CSF M-CSF + Pam 3 CSK ** * M-CSF + LPS M-CSF + b-glucan particles M-CSF + C. albicans 5 0 ** 5 0 Day 7 Pam 3 CSK 4 Day 7 LPS Gil et al. unpublished
20 Effects of UV and PGE2 on HSPCs Prue Hart, Telethon Kids Institute, Perth, Australia UV exposure => immunosuppressive; inefficient T cell priming by DC cells; PGE2-mediated effect persists following bone marrow transplantation bmdcs migration of adoptively transferred bmdcs to draining lymph nodes => inefficient T cell priming Scott et al. (2014) Journal of Leukocyte Biology 95:225-32
21 Effects of UV and PGE2 on HSPCs Prue Hart, Telethon Kids Institute, Perth, Australia UV/PGE2 effect persists upon serial bone marrow transplantation bmdcs bmdcs bmdcs => contact hypersensitivity assay Scott et al. (2014) Journal of Leukocyte Biology 95:225-32
22 Effects of UV and PGE2 on HSPCs - reduced myeloid cell migration Prue Hart, Telethon Kids Institute, Perth, Australia macrophage and neutrophil migration also reduced Ng et al. (2013) Journal of Immunology 190: Scott et al. (2014) Journal of Leukocyte Biology 95:225-32
23 Summary Hematopoietic stem and progenitor cells possess the machinery to detect and respond to cytokines, microbial products, damage etc. Endogenous factors and microbial products regulate innate immune cell differentiation in the steady-state and during emergency myelopoiesis Epigenetic modifications propagated through myelopoiesis have the potential to impact innate immune cell function for long periods
24 Potential implications for non-specific effects of vaccination Mechanism to extend innate immune memory? Positive v. negative effects? Live v. inactivated vaccines detected differently? Direct v. indirect detection by HSPCs? Programming of NK cells/other innate lymphoid cells? Early v. later life effects, immunosenescence? Role for microbiome?
25 Goodridge Lab Nargess Hassanzadeh-Kiabi Shane Sakamaki-Ching Alberto Yáñez Madelena Ng Aparna Subramanian University of Valencia, Spain Javier Megías Alba Martínez Daniel Gozalbo María Luisa Gil
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