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Adaptive immunity 2 - Immune regulatory mechanisms through B cell axis

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Immune regulatory mechanisms through B cell axis

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Adaptive immunity 2 - Immune regulatory mechanisms through B cell axis

  1. 1. ©2015 Osaka University. All rights reserved. Immune regulatory mechanisms through B cell axis Tomohiro Kurosaki Osaka University Adaptive Immunity 2 Number 1 Number 2 Number 3 Antigen presenting cell Helper T cell inflammatory cytokine (IL-6,M1P1α,M1P1) inflammatory cytokine T cell activation B cell 1
  2. 2. ©2015 Osaka University. All rights reserved. Inflammatory cytokine (IL-6,M1P1α,M1P1) Anti-inflammatory cytokine (IL-10) Breg cells in mice 2
  3. 3. ©2015 Osaka University. All rights reserved. DTH: Delayed-type hypersensitivity EAE: Experimental autoimmune encephalomyelitis IBD: Inflammatory bowel diseases MS: Multiple sclerosis SLE: Systemic lupus erythematosus Year Mouse Human 1974 B cells suppress DTH (Katz SI et al., Nature) 1996 B cell-deficient mice develop EAE (Wolf SD et al., J. Exp. Med.) 2002 IL-10+/+ B cells suppress EAE (Fillatreau S et al., Nature Immunol.) B cells control IBD (Mizoguchi A et al., Immunity) 2007 B cells have an impaired IL-10 production in MS (Duddy M et al., J. Immunol.) 2010 B cells have an impaired IL-10 production in SLE (Blair PA et al., Immunity) Highlights in regulatory B cell research DTH: Delayed-type hypersensitivity EAE: Experimental autoimmune encephalomyelitis IBD: Inflammatory bowel diseases MS: Multiple sclerosis SLE: Systemic lupus erythematosus Year Mouse Human 1974 B cells suppress DTH (Katz SI et al., Nature) 1996 B cell-deficient mice develop EAE (Wolf SD et al., J. Exp. Med.) 2002 IL-10+/+ B cells suppress EAE (Fillatreau S et al., Nature Immunol.) B cells control IBD (Mizoguchi A et al., Immunity) 2007 B cells have an impaired IL-10 production in MS (Duddy M et al., J. Immunol.) 2010 B cells have an impaired IL-10 production in SLE (Blair PA et al., Immunity) Highlights in regulatory B cell research 3
  4. 4. ©2015 Osaka University. All rights reserved. Wolf SD et al. J. Exp. Med. 184: 2271-2278,1996 Data from individual mice were plotted from three consecutive B10.PL (squares) (B) and four consecutive B10.PLμMT (circles) (C) mice. The data was collected from one fifth of the experiments set up. DiseaseSeverity DAY DTH: Delayed-type hypersensitivity EAE: Experimental autoimmune encephalomyelitis IBD: Inflammatory bowel diseases MS: Multiple sclerosis SLE: Systemic lupus erythematosus Year Mouse Human 1974 B cells suppress DTH (Katz SI et al., Nature) 1996 B cell-deficient mice develop EAE (Wolf SD et al., J. Exp. Med.) 2002 IL-10+/+ B cells suppress EAE (Fillatreau S et al., Nature Immunol.) B cells control IBD (Mizoguchi A et al., Immunity) 2007 B cells have an impaired IL-10 production in MS (Duddy M et al., J. Immunol.) 2010 B cells have an impaired IL-10 production in SLE (Blair PA et al., Immunity) Highlights in regulatory B cell research 4
  5. 5. ©2015 Osaka University. All rights reserved. Fillatreau S. et al., Nature Immunol. 10, 944-950, 2002 IL-10-producing B cells suppress EAE development Clinicalscore Time (d) IL-10-/- B cells B6 B cells T2-MZP BREG cell B10 cell IgMhigh IgMhigh CD21high CD21high CD24high CD24high CD1dhigh CD1dhigh CD19CD19 5
  6. 6. ©2015 Osaka University. All rights reserved. DTH: Delayed-type hypersensitivity EAE: Experimental autoimmune encephalomyelitis IBD: Inflammatory bowel diseases MS: Multiple sclerosis SLE: Systemic lupus erythematosus Year Mouse Human 1974 B cells suppress DTH (Katz SI et al., Nature) 1996 B cell-deficient mice develop EAE (Wolf SD et al., J. Exp. Med.) 2002 IL-10+/+ B cells suppress EAE (Fillatreau S et al., Nature Immunol.) B cells control IBD (Mizoguchi A et al., Immunity) 2007 B cells have an impaired IL-10 production in MS (Duddy M et al., J. Immunol.) 2010 B cells have an impaired IL-10 production in SLE (Blair PA et al., Immunity) Highlights in regulatory B cell research Duddy M et al. J Immunol. 178: 6092-6099, 2007 IL-10 Cytokinesecretion(pg/ml) CD40 alone Dual Stimulation p=0.008 p=0.037 Normal Multiple Sclerosis 6
  7. 7. ©2015 Osaka University. All rights reserved. CD19+CD24hiCD38hi B cells from SLE patients express lower amounts of IL-10 CD24hi CD38hi CD24hi CD38hi CD24hi CD38int CD24hi CD38- CD24hi CD38- CD24hi CD38int Blair P.A. et al, Immunity 32, 129-140, 2010 Healthy SLE CD19+CD24hiCD38hi B cells from healthy human individuals suppress T helper cell differentiation through IL-10 :CD24hiCD38hi B Cells Blair P.A. et al, Immunity 32, 129-140, 2010 7
  8. 8. ©2015 Osaka University. All rights reserved. Th17 cells Th1 cells Antigen TLR ligand TLR BCR CD40 MHCII TCR B10 cells Activated B10 cells CD4+ T cells CD40 IL-10 IL-17 IFN-γ CD40L MHCII The model of regulatory B cell development Mauri, C. et al., Annu. Rev. Immunol., 30:221-241, 2012. Yoshizaki, A., et al., Nature, 491:264-268, 2012. Our working model 8
  9. 9. ©2015 Osaka University. All rights reserved. What lineage of B cell is producing IL-10 in vivo ? Lymph node B cells FAS+GL7+B cells (GC B) CD138+CD44+ cells are main IL-10-producing B cells IL-10-Venus B220+ cells CD138+CD44+ cells IL-10-Venus Naïve mice (day 0) EAE-induced mice (days 14) B220+ cells WT mice IL-10-Venus mice Matsumoto M. et al, Immunity 41, 1040-1051, 2014 9
  10. 10. ©2015 Osaka University. All rights reserved. Draining LN cells 0 2 4 6 8 10 12 days 0 days 7 days 14 days 21 days 28 Cells(x104) Gated on CD138+CD44+ cells Blimp1-GFP EAE-induced Blimp1-GFP mice CD138 Plasmablast (Blimp1int) CD138+CD44+ cells in dLN are plasmablasts Matsumoto M. et al, Immunity 41, 1040-1051, 2014 Oracki, S. A. et al., Immunol. Review, 237:140-159, 2010 Blimp1 is a master gene to differentiate into CD138+ plasmablasts and plasma cells Activated B cell Plasmablast Short-lived Plasmablast Long-lived Plasmablast Surface lg+ B220+ Synd-1- Flt3+ MHC ll++ CXCR4- Pax5+++ Irf4+ Surface lg+ B220 lo Synd-1+ Flt3- MHC ll+ CXCR4+ Pax5- Irf4+++ Blimp-1+ Surface lg- B220- Synd-1+ Flt3- MHC ll- CXCR4+++ Pax5- Irf4+++ Blimp-1+++ Surface lg- B220- Synd-1+ Flt3- MHC ll+ CXCR4++ Pax5- Irf4+++ Blimp-1++ (CD138+) 10
  11. 11. ©2015 Osaka University. All rights reserved. EAEscore Days Blimp1 BKO mice exhibit increased severity of EAE 0 1 2 3 4 5 0 10 20 30 MBC mbprdm1 P<0.05 Blimp1 BKO Control Matsumoto M. et al, Immunity 41, 1040-1051, 2014 Where can B cells suppress EAE ? 11
  12. 12. ©2015 Osaka University. All rights reserved. dLNs zz CD138+ Medullary sinus Is the spleen a suppression site? Splenectomy Spleen CD138+ VeinArtery VeinArtery 24 Splenectomy mice show normal EAE development 0 1 2 3 4 5 0 10 20 30 Sham Splenectomy EAEscore Days Matsumoto M. et al, Immunity 41, 1040-1051, 2014 12
  13. 13. ©2015 Osaka University. All rights reserved. zz Medullary sinus CD138+ WT VeinArtery CD138+ WT HEV B cell-deficient mice Are LNs a suppression site? WT B cells Artery Vein Spleen dLNs zz Medullary sinus KO CD138+ KO HEV B cell-deficient mice Are LNs a suppression site? L-selectin KO B cells VeinArtery VeinArtery Spleen dLNs 13
  14. 14. ©2015 Osaka University. All rights reserved. 0 1 2 3 4 5 0 10 20 30 WT B cells L-sel KO B cellsEAEScore Days L-sel KO B cells do not suppress EAE development Matsumoto M. et al, Immunity 41, 1040-1051, 2014 How does B cell IL10 suppress EAE ? 14
  15. 15. ©2015 Osaka University. All rights reserved. Th17 cells B10 cells CD4+ T cells IL-10 TLR BCR CD40 MHCIITCR CD40L Dendritic cell Plasmablast MHCII Naive T cells Th1 cells IL-10R Antigen TCR The model of regulatory function of plasmablasts CD4+ T cells DCCD8+ T cells B cells Isotype control Anti-IL-10R DC in the draining LN express IL-10 receptor EAE-induced WT mice (days 14) IL-10R Th1 cells Th17 cells IL-10R Matsumoto M. et al, Immunity 41, 1040-1051, 2014 15
  16. 16. ©2015 Osaka University. All rights reserved. 0 200 400 600 800 1000 1200 1400 LPS LPS+IL-10 IL-10 can inhibit the production of IL-6 and IL-12 by DC IL-6(pg/ml) LPS LPS + IL-10 LPS LPS + IL-10 IL-6 IL-6 Dendritic Cell (DC) Dendritic Cell (DC) Matsumoto M. et al, Immunity 41, 1040-1051, 2014 TGF- + MOG35-55 IL-10 can inhibit the differentiation of naïve T cells into Th17 cells IFN- IL-17 IFN- IL-17 LPS LPS + IL-10 IL-6 IL-6DC Naïve T cells MOG TCR Matsumoto M. et al, Immunity 41, 1040-1051, 2014 16
  17. 17. ©2015 Osaka University. All rights reserved. Th17 cells B10 cells CD4+ T cells IL-10 TLR BCR CD40 MHCIITCR CD40L Dendritic cell Plasmablast MHCII Naive T cells Th1 cells IL-10R Antigen TCR The model of regulatory function of plasmablasts Artery Vein Afferent lymphatic vessels Medullary sinus Girard, J.P. et al., Nature Rev. Immunol., 12:762-773, 2012. Wehrlii, N., et al., Eur. J. Immunol., 31:609-616, 2001. Dendritic Cell Efferent lymphatic vessels HEV B T PB T B 17
  18. 18. ©2015 Osaka University. All rights reserved. Plasmablasts are mainly localized in the T-B border area B B T Plasmablast (CD138) DC (CD11c) Merge B B T B B T Efferent lymphatic Medullary sinus Afferent lymphatic B cell follicle T cell area Matsumoto et. al. Immunity 41, 1040-1051, 2014 Central nervous system (CNS) CNS autoantigen Neuron Myelin sheath CNS autoantigen (MOG35-55) Blood-brain barrier IFN- IL-17 Subcutaneous tissue Lymph node CD4+ T cells B cell follicle DC DC EAE, a mouse model of multiple sclerosis, is caused by autoreactive T cells Blood circulation Tfh Th17 Th1 Blood circulation 18
  19. 19. ©2015 Osaka University. All rights reserved. CD4+ T cells Tfh Th17 Th1 B cell follicle PB Dendritic cell B cell follicle Lymph node Tfh Tfh CD4+ T cells Dendritic cell BCR Antigen Naïve B cells Activated B cells PB Tfh The model of regulatory function of plasmablasts Central nervous system (CNS) CNS autoantigen Neuron Blood circulation CNS autoantigen (MOG35-55) Blood-brain barrier Subcutaneous tissue Th17 B cell follicle IFN- IL-17 IL-10 TregTreg Th1 Th17 Th1 DC DC EAE, a mouse model of multiple sclerosis, is caused by autoreactive T cells Myelin sheath 19
  20. 20. ©2015 Osaka University. All rights reserved. What are essential factors for IL-10 production inside B cells? IRF4 is essential for NFAT-dependent IL-10 production B cells Plasmablasts BCR BCRTLR Differentiation TLR Irf4NF-B IRF4 NFAT Il-10 20
  21. 21. ©2015 Osaka University. All rights reserved. IRF4 expression is induced by TLR stimulation Day 0 Days 2 Days 4 WT IRF4 -actin LPS 48 hr WB (IRF4 and -actin) Naive B cells Matsumoto M. et al, Immunity 41, 1040-1051, 2014 IRF4 is essential for IL-10 production IL-10(pg/mlper105cells) 0 2000 4000 6000 8000 10000 No anti-IgM PI WT IRF4 KO LPS + BCR stimulation LPS ELISA LPS 48 hr BCR stimulation 24 hr PBPBB Matsumoto M. et al, Immunity 41, 1040-1051, 2014 21
  22. 22. ©2015 Osaka University. All rights reserved. IRF4 BKO mice show increased severity of EAE EAEScore Days 0 1 2 3 4 5 6 0 7 14 21 28 35 Control IRF4 BKO Matsumoto M. et al, Immunity 41, 1040-1051, 2014 1000 IL-10(pg/mlper10 5 cells) 0 500 2000 1500 STIM1&2 BKO Control STIM KO plasmablasts impair IL-10 production LPS + BCR stimulation LPS ELISA LPS 48 hr BCR stimulation 24 hr PBPBB Matsumoto M. et al, Immunity 41, 1040-1051, 2014 22
  23. 23. ©2015 Osaka University. All rights reserved. Control STIM1 BKO Control STIM2 BKO Control STIM1&2 BKO 2mM Ca2+ Time (sec) anti-IgM EGTA Fluorescenceratio Spleen: B220+ gate Control: mb1-Cre mice anti-IgM anti-IgM Ca2+ Ca2+ Ca2+ Calcium influx in STIM-deficient B cells Matsumoto M. et al, Immunity 34, 703-714, 2011 0 1 2 3 4 5 0 10 20 30 Clinicalscore P < 0.05 Time (d) STIM1&2 BKO Control STIM-deficiency exacerbates the development of EAE Matsumoto M. et al, Immunity 34, 703-714, 2011 23
  24. 24. ©2015 Osaka University. All rights reserved. IRF4 is essential for NFAT-dependent IL-10 production B cells Plasmablasts 47 BCR BCRTLR Differentiation TLR Irf4NF-B IRF4 NFAT Il-10 How about humans? 24
  25. 25. ©2015 Osaka University. All rights reserved. None IL-2/6/IFN CpG+IL-2/6/IFNCpG CD38 CD27 CD27int plasmablast Naïve B Memory B CD27hi plasmablast Concomitant treatment with CpG and cytokines induces the generation of CD27intCD38+ cells Matsumoto M. et al, Immunity 41, 1040-1051, 2014 Naïve B Memory B CD27int plasmablast CD27hi plasmablast CD27intCD38+ cells consist of plasmablasts Matsumoto M. et al, Immunity 41, 1040-1051, 2014 25
  26. 26. ©2015 Osaka University. All rights reserved. 0 200 400 600 800 0 2000 4000 6000 8000 0 2000 4000 6000 8000 10000 IL-10(pg/ml) None Anti-IgM PMA+Ionomycine CD27int plasmablasts selectively secrete IL-10 Matsumoto M. et al, Immunity 41, 1040-1051, 2014 0 500 1000 1500 CD38 CD27 Naïve B Memory B IL-10(pg/ml) IL-10-producing CD27int plasmablasts are derived from naïve B cells Culture with CpG+IL-2/6/IFN CD27 Matsumoto M. et al, Immunity 41, 1040-1051, 2014 26
  27. 27. ©2015 Osaka University. All rights reserved. IL-10-producing CD27int plasmablasts are derived from naïve B cells IL-10 Cytokine receptors BCR BCRTLR Differentiation TLR Naive B cells TLR Memory B cells Differentiation BCRTLR CD27hi plasmablasts CD27int plasmablasts Conclusions 1) Plasmablasts in LNs produce IL-10 to suppress EAE 2) Plasmablasts move across interfollicular and T cell zones, probably interacting with DCs 3) IL-10 inhibits DC functions to generate Th17 cells 4) The above mechanism is probably operating in humans → Plasmablasts control T-cell autoimmunity through their IL-10 production 54 27

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