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The Advances of regulatory dendritic cells in The Advances of regulatory dendritic cells in GVHD and GVL after allogeneic bone marrow GVHD and GVL after allogeneic bone marrow
transplantationtransplantation
Yihong Huang, Kailin Xu ( 黄一虹,徐 林开 )
Institute of HematologyThe Affiliated Hospital of Xuzhou Medical College
(徐州医学院血液病研究所)
OutlineOutline
Ⅱ. Dendritic cells and regulation of GVHD and GVL
Ⅰ . The Immunological Basis for GVHD and GVL
Ⅲ. New Exploration to Modulate DCs in Our Lab
The classical Model of the Immunopathogenesis of GVHD Described by Three Interlinked Phases
Pro-inflammatory environment
Donor T cell proliferation after encountering activated
recipient APCs bearing alloantigen
Alloreactive cell migration to target tissues and resulting in
tissue damage in the form of GVHD
Ferrara JT, et al. Stem Cells. 1996,14:473-489.
Ferrara JT, et al. Semin Hematol.2006,43:3-10
Immunopathogenesis of Alloreactive Response and Development of GVHD
Immunopathogenesis of Alloreactive Response and Development of GVHD
Immunopathogenesis of Alloreactive Response and Development of GVHD
New Approaches to Separate GVHD and
GVL Effects
Novel approaches to modify the pro-inflammatory environment
New exploration to modulate donor T cell priming and expansion
New ways to manipulate donor T cell migration to target tissues
Novel approaches to modify the pro-inflammatory environment
New exploration to modulate donor T cell priming and expansion
New ways to manipulate donor T cell migration to target tissues
APC involved
Pan B, Xu KL et al. Cytokine.2014, 68(2):69-75 Li ZY, Xu KL et al. Hematology. 2013,18(6):352-359 Zhao K, Xu KL et al. Immunobiology. 2013;218:1505-13Zeng LY, Xu KL et al. Int J Hematol. 2012,95(2):189-97
OutlineOutline
Ⅱ. Dendritic cells and regulation of GVHD and GVL
Ⅰ . The Immunological Basis for GVHD and GVL
Ⅲ. New Exploration to Modulate DCs in Our Lab
Ⅱ. DCs and regulation of GVHD and GVL
2. The role of DCs in GVHD and GVL
3. Manipulated DCs for prevention or treatment of GVHD
4. Generation of tolDCs for prevention or treatment of GVHD
5. Cell therapies that target DCs in vivo
6. Active clinical trials that target/impact DCs
1. Overview of DCs subsets
Overview of DC subsetsBased on the source of DC Based on the function of DC
Mature DCs(mDCs): enhance T cell immune response;
ImmatureDCs (iDCs): involved in the induction of peripheral T cell tolerance under steady state conditions ,but likely change into mature under inflammatory conditions;
Regulatory DCs (rDCs)/Tolerogenic DCs(tolDCs): established DCs with potent immunoregulatory property, to impair the functions of alloreactive/xeno reactive T cells.
Elizabeth O. Stenger et al. Blood. 2012;119(22):5088-5103
Ⅱ. DCs and regulation of GVHD and GVL
1. Overview of DCs subsets
2. The role of DCs in GVHD and GVL
3. Manipulated DCs for prevention or treatment of GVHD
4. Generation of tolDCs for prevention or treatment of GVHD
5. Cell therapies that target DCs in vivo
6. Active clinical trials that target/impact DCs
The Important role of Dendritic Cells in GVHD and GVL
Acute GVHD can be induced by either host or donor APCs, whereas
host APCs are required for the initiation of aGVHD and donor APCs
amplify the process.
Dendritic cells(DCs) as professional APCs, have been well recognized
for their role in the induction of GVHD and GVL responses .
Traditional therapies have targeted T cells, yet immunostimulatory
DCs are critical in the pathogenesis of GVHD.
Zhao K, Xu KL et al. J Interferon Cytokine Res. 2014,34(9):707-15 . Pan B, Xu KL et al. Clinical Immunology (2014) 150, 101–8.Sang W, Xu KL et al. Immunol Lett . 2011,136(2):194-202 .
The role of DCs in GVHD
Initial tissue injury may trigger DCs within tissues and induce DCs to migrate to draining lymph nodes;
Migrating DCs “trap” autoreactive T cells , where they induce T cell activation and may “imprint” a homing phenotype that permits selective trafficking of T cells to GVHD target organs;
DCs within inflamed tissues may act to amplify the developing GVHD by providing further priming signals to T cells.
DCs can influence development of GVHD at multiple levels:
Ronjon Chakraverty et al. Blood. 2007;110:9-17
The role of DCs in GVL
Host DCs are required for full GVL effects, although donor DCs can initiate GVL activity when low burden of tumor are present..
Depletion of donor BM CD11b myeloid cells in an experimental model enhanced survival of recipients with tumor;
Addition of precursor pDCs, augmented GVL without increase in GVHD;
Increased graft pDC is associated with relapse and decreased patient overall survival (OS).
Li JM et al .Biol Blood Marrow Transplant. 2004;10(8):540-551.
Li JM et al. Immunol. 2009;183(12):7799-7809.
Rajasekar R et al.Biol Blood MarrowTransplant. 2010;16(6):854-860.
Ⅱ. DCs and regulation of GVHD and GVL
1. Overview of DCs subsets
2. The role of DCs in GVHD and GVL
3. Manipulated DCs for prevention or treatment of GVHD
4. Generation of tolDCs for prevention or treatment of GVHD
5. Cell therapies that target DCs in vivo
6. Active clinical trials that target/impact DCs
Manipulated DCs for prevention or treatment of GVHD
DCs may be targeted using antibodies to cell surface molecules or can be manipulated in vitro to skew their function to suppress GVHD.
Pharmacologic interventions Biologic interventions Vaccination strategies
Manipulated DCs in HSCTTable 1. Impact of interventional stategies on DCs and outcome in experimental HSCT
Elizabeth O. Stenger et al. Blood. 2012;119(22):5088-5103
Manipulated DCs in HSCTTable 2. Overview of DC vaccination trials after allo-HSCt
Maud Plantinga et al. Frontiers in immunology. 2014,5:218
Ⅱ. DCs and regulation of GVHD and GVL
1. Overview of DCs subsets
2. The role of DCs in GVHD and GVL
3. Manipulated DCs for prevention or treatment of GVHD
4. Generation of tolDCs for prevention or treatment of GVHD
5. Cell therapies that target DCs in vivo
6. Active clinical trials that target/impact DCs
Generation of tolDCs for prevention or treatment of GVHD
Immature or maturation-resistant tolDCs can promote tolerance in experimental organ and HSCT, while still protecting against leukemia relapse.
DCs can be manipulated in vitro to produce tolDCs:
• Cytokine growth factors• Genetic engineering• Phamacological mediators
Generation of tolerogenic DC in vitro
Usage of tolDC therapy to inbibit GVHD in miceTable 3. Summary of tolDC used in allo-HSCT
Elizabeth O. Stenger et al. Blood. 2012;119(22):5088-5103
Ⅱ. DCs and regulation of GVHD and GVL
1. Overview of DCs subsets
2. The role of DCs in GVHD and GVL
3. Manipulated DCs for prevention or treatment of GVHD
4. Generation of tolDCs for prevention or treatment of GVHD
5. Cell therapies that target DCs in vivo
6. Active clinical trials that target/impact DCs
Mesenchymal stem cells (MSCs): have shown promise in the prevention and treatment of GVHD.
• Human MSCs impair DC maturation and induce T-cell hyporesponsiveness.
• TolDCs generated by coculture of DCs with human MSCs (MSC-DCs) induce Ag specific Treg via activation of the Notch pathway.
Cell therapies that target DCs in vivo
Myeloid-derived suppressor cells (MDSCs) : modulate both innate and adaptive immunity with immunosuppressive properties.•MDSCs inhibit the differentiation and maturation of DCs.
•MDSCs were more potent inhibitors of MHC-mismatched GVHD.
Perla Filippini et al. Expert Rev. Clin. Immunol. 2014,10(10), 1357–1374
Chen W, Xu KL et al. Immunol Lett. 2012,143(2):161-9.Chen W, Xu KL et al. Cell Biochem Biophys. 2013, 67:1181–1191 .
Cell therapies that target DCs in vivo
Regulatory T cells(Treg):
• Treg therapy supports tolerance through effects on DCs, while DCs
also control the number and function of Treg.
• Human Treg can abrogate GVHD via induction of PD-L1 expression
on conditioned DCs and on effector T cells.
• Ag-specific Treg can be induced and expanded by monDCs in an
IDO-dependent manner.
Cao J, Xu KL et al. Leuk Res. 2010,34(10): 1374-82Chen C, Xu KL et al. Transplant Proc. 2011,43(5): 2041-8 .Perla Filippini et al. Expert Rev. Clin. Immunol. 2014,10(10), 1357–1374
DC based methods of transplant tolerence
Adrian E. Morelli et al. Nature review immunology. 2007,7:610-321
Ⅱ. DCs and regulation of GVHD and GVL
1. Overview of DCs subsets
2. The role of DCs in GVHD and GVL
3. Manipulated DCs for prevention or treatment of GVHD
4. Generation of tolDCs for prevention or treatment of GVHD
5. Cell therapies that target DCs in vivo
6. Active clinical trials that target/impact DCs
There are numerous open clinical trials for the prevention
or treatment of GVHD currently studying pharmacologic or
biologic interventions and cellular therapies that target or
impact DCs.
Active clinical trials that target/impact DCs
Table 4. Active clinical trials using interventions that target/ impact DCs
Elizabeth O. Stenger et al. Blood. 2012;119(22):5088-5103
OutlineOutline
Ⅱ. Dendritic cells and regulation of GVHD and GVL
Ⅰ . The Immunological Basis for GVHD and GVL
Ⅲ. New Exploration to Modulate DCs in Our Lab
Ⅲ. New Exploration to Modulate DCs in Our Lab
1. Effects of regulatory dendritic cells on GVHD and GVL in mice after
allogeneic bone marrow transplantation
2. Separation of GVHD and GVL effects in mouse allo-HSCT model by
engineered DCs
Part 1:
Effects of regulatory dendritic cells on GVHD and GVL in mice after
allogeneic bone marrow transplantation
Generation of inducible DCreg
A, imDC ; B, mDC ; C, DCreg ; D, LPS stimulated regDC
Figure 1. The cell morphological observation of DC subsets
Compared with mDCs, shorter synapsis was shown in DCreg; and rare wrinkles were observed in DCreg.
Surface molecules expression on DCs
mDCimDC DCreg
Co-stimulatory molecules were down regulated in DCreg
Figure 2. FACS was used to detected the molecules of expressed on DCs
Mice survival after transplantation
The first death in the allo-BMT group was found on 10 d, and all mice died within 25 d;
Most of the mice in the imDC group died within 21 d, while 20% of the mice were still alive at 30 d;
The first death in the DCreg group was found on 19 d, while 46.7% of the mice were still alive at 30 d.
Figure 3. The survival rate of mice with allo-BMT
Chen C, Xu KL. Leukemia Research, 2014 , 38(12):1460-8
Effects of DCreg on pathological changes in GVHD tissues after transplantation
Liver cell edema, focal necrosis, lymphocyte infiltration at the portal area, intestinal epithelial cells necrosis, significant inflammatory cell infiltration in the pulmonary vessels in the allo-BMT and imDC groups;
Mice in the DCreg group, we found only mild or even no pathological damage in target organs.
Normal allo-BMT imDC DCreg
Liver
Gut
Lung
Figure 4. Target organs were detected after HE staining
Pathological GVHD score was low in DCreg group
Groups Liver Intestine
Lung
allo-BMT 6.33±0.58 7.00±1.00 6.67±0.58
imDC 4.67±0.58* 4.67±0.58* 2.33±1.16*
DCreg 3.00±1.00*# 2.67±0.58*# 2.00±1.00*#
Table 1. GVHD pathologic scoring of liver, small intestine, and lung in mice after transplantation (n = 3, )sx ±
*P < 0.05 vs allo-BMT group; #P < 0.05 vs imDC group
Cytokine levels in plasma
IFN-γ levels increased significantly in the allo-BMT group, whereas low level of that was detected in DCreg group.
IL-10 as immune surppressor cytokine increased obviously in Dcreg mice.
Figure 5. cytokines from allo-BMT mice were detected by ELISA
Part 2:
Suppression of graft-vs-host disease and retention of graft-vs- leukemia reaction by murine genetic engineering dendritic cells after bone marrow transplantation
Establishment of engineered DCs
Lentivirus-mediated expression of soluble tumor necrosis factor
receptor 1 (sTNFR1) converted immature DC were established in
vitro, named sTNFR-DC;
PXZ9-DC were used as negetive vector control;
Mice survival following transplantation
Mice from the leukemia model group began to die after 13 days, and all died due to leukemia within 18 days;
allo-BMT group began to die after 8 days, and all died by 18 days
Most of the pXZ9-imDC group mice died within 21 days
sTNFR1-imDC group mice died starting at 15 day, and 40% of mice survived up to 30 days
Figure 6. The survival rate of mice after allo-BMT
Pan B, Xu KL. Immunol Lett . 2012,142(1-2):48-54
Evaluation of GVHD
High GVHD clinical score were evaluated in allo-BMT mice , whereas injected sTNFR-DC mice showed slight GVHD score.
More severe pathological damage were observed in allo-BMT group, but mild changes were shown in sTNFR-DC mice.
Skin
Liver
Gut
Allo-BMT sTNFR1-DC PXZ9-DC
Figure 6. The clinical score and pathological changes in mice
Cytokine levels in plasma
IFN-γ levels increased significantly in the allo-BMT group and PXZ9-DC group, whereas low level of IFN-γ was detected in sTNFR-DC group.
IL-4 which was considered as immune surppressor cytokine increased obviously in sTNFR-DC group.
Figure 7. Cytokine expression in mice after allo-BMT
Prospective
Synergistic therapies or those that
target both T and DC cells may be
more effective.
The injection site, dose and frequency
of the tolDCs induced in vitro should be
carefully considered.
Potential DC-based therapies for GVHD :
Thank You for Your Attention