scholarly journals The IL-27/Wsx-1 Axis Modulates T Cell Responses and Regulates Acute Graft-Versus-Host Disease after Allogeneic Bone Marrow Transplantation

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 2146-2146
Author(s):  
Shoubao Ma ◽  
Huanle Gong ◽  
Shuangzhu Liu ◽  
Jingjing Han ◽  
Zhinan Yin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Cd8 T Cells ◽  
Acute Gvhd ◽  
Treg Cells ◽  
Graft Versus Host ◽  
Cell Responses ◽  
Target Organs ◽  
Ifn Γ

Abstract IL-27 is a member of the IL-12 cytokine family that consists of EBV-induced gene 3 (EBI3), and p28. IL-27 is produced by activated antigen-presenting cells such as dendritic cells (DCs) and macrophages, and it signals through a heterodimeric receptor (IL-27R) consisting of the WSX-1 and the gp130. Emerging evidence has shown that IL-27 has both pro- and anti-Inflammatory effects. Recent study demonstrated a protected role IL-27p28 in acute graft-versus-host disease (aGVHD) in parent-to-F1 murine aGVHD model (Marillier et al., Eur. J. Immunol. 2014). However, the precise role of IL-27 in the development of aGVHD remains largely unknown. In this study, we carried out studies in a murine aGVHD model of fully MHC-mismatched myeloablative bone marrow transplantation (C57BL/6 to BALB/c). Lethally irradiated BALB/c mice transplanted with WT B6 bone marrow (BM) plus IL-27-/-(IL-27p28flox/flox-CD11c-cre (Itgax-p28f/f)) splenocytes had significantly accelerated aGVHD mortality comparing with those received WT BM plus splenocytes (P<0.001). However, similar aGVHD mortality was observed between mice transplanted with WT BM plus WT splenocytes and IL-27-/-BM plus WT splenocytes, and IL-27-/-BM plus IL-27-/- splenocytes, suggesting that IL-27 from splenocytes, not BM-derived cells, had protective effect in aGVHD. Additionally, mice were transplanted with IL-27-/- BM and splenocytes, and then treated with rmIL-27 on days 0 and 7 post BMT. rmIL-27 treated mice had longer survival and alleviated the clinical signs of aGVHD compared to PBS-treated mice (P<0.05). We next explored the mechanisms by which IL-27 protects mice against aGVHD. We found the expression of the cell surface IL-27 receptor, WSX-1 and gp130, were increased on both CD4+ and CD8+T cells after allo-stimulation. Moreover, IL-27 inhibited cell proliferation of allo-reactive T cells in mix lymphocytes reactivation (MLR) in vitro. We also found that the percentages of CD69+CD4+, CD69+CD8+T cells, as well as IFN-γ expression by CD4+ and CD8+T, were significantly increased in the spleen, liver and intestinal intraepithelial lymphocytes (IEL) from mice transplanted with WT BM and IL-27-/- splenocytes. Furthermore, we observed increased frequencies and numbers of Treg cells and MDSCs in aGVHD target organs in mice reconstituted with IL-27-/-splenocytes. Together, these results indicated that IL-27 alleviated aGVHD may through suppressing Th1 cell responses and promoting immune suppressing cells, including MDSCs and Treg cells. To further clarify the role of IL-27 in aGVHD, mice were transplanted with WT BM plus IL-27R-/-(B6N.129P2-Il27ratm1Mak/J) splenocytes, the results, however, showed that IL-27R deficiency in donor T cells significantly attenuated aGVHD, which was unpredicted, and was contrary to that of IL-27 deficiency results. Additional studies showed that IL-27R deficiency in T cells inhibited allo-reactive T cells proliferation and IL-2, IFN-γ production in MLR assay. The percentages of CD69+ T cells and IFN-γ+ CD4+ and CD8+T were significantly decreased, while the MDSCs and Treg cells wereincreased in aGVHD target organs from mice transplanted IL-27R-/-splenocytes. These results indicated that lack of IL-27R signaling resulted in downregulation of intrinsic immune responses which led to alleviation of aGVHD. Previous study reported that soluble form of IL-27Ra is highly existed in human serum and may severs as a natural IL-27 antagonist (Dietrich et al., JI. 2014). We hypothesized that sIL-27Ra may function as a decoy receptor to regulate aGVHD through inhibiting IL-27 signaling. To test this hypothesis, WT recipients were given exogenous soluble mouse IL-27 Ra Fc chimera protein known to block the binding of IL-27 to the membrane bound form of IL-27R. WT recipients given IL-27 Ra-Fc to block IL-27R/IL-27 interaction had significantly exacerbated GVHD mortality. The percentages of CD69+ T cells and IFN-γ+ CD4+ and CD8+T were significantly increased in aGVHD target organs from mice received IL-27 Ra fusion protein. In conclusion, this is the first demonstration in the same allogeneic BMT model that IL-27 deficiency augments but IL-27R deficiency alleviates acute GVHD. Our studies provide further evidence for the protective role of rmIL-27, and promoting effect of sIL-27R in the same aGVHD model. This study warrants further investigations of the possible therapeutic applications of IL-27 in acute GVHD. Disclosures No relevant conflicts of interest to declare.

T Cell Activation and Regulation in Graft-Versus-Host Disease: Integral Role of CD28, CTLA4 and GITR Splice Variants.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3054-3054
Author(s):  
Yuji Miura ◽  
Christopher J. Thoburn ◽  
Emilie C. Bright ◽  
Elizabeth C. Matsui ◽  
William H. Matsui ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Activation ◽  
Cd8 T Cells ◽  
Cell Activation ◽  
Splice Variants ◽  
Treg Cells ◽  
Mrna Splice ◽  
Graft Versus Host

Abstract Graft-versus-host disease (GVHD) is a serious, life-threatening complication that occurs following allogeneic (allo) bone marrow transplantation (BMT). The use of non-specific immunosuppression or T cell depletion has reduced the incidence of GVHD but at the expense of increased rates of infection and leukemic relapse. Modulation of the major costimulatory pathway (CD28/CTLA4:B7) involved in T cell activation and regulation may lead to specific immune tolerance in the absence of global non-specific immunosuppression. The identification of mRNA splice variants encoding for soluble forms of CD28, CTLA4 and GITR suggests that costimulation of T cells is complex and is not limited to cell-cell contact. The present studies examined the hypothesis that the onset of GVHD and the re-establishment of immune tolerance correlate with the expression levels of these costimulatory molecules. mRNA transcript levels for the soluble (s) and full-length (fl; cell surface associated) variants assessed by quantitative PCR, were temporally examined in peripheral blood lymphocytes (PBLs) from patients undergoing alloBMT (n=38) or autologous (auto) BMT (n=39) with the induction of autoGVHD by cyclosporin A treatment post-transplant. Levels of s and fl CD28 mRNA transcripts in PBLs were significantly increased (&gt;1.5 fold, P&lt;0.05) in patients developing either allo or autoGVHD compared to patients who do not develop GVHD. s and flCTLA4 levels in patients at the onset of allo and autoGVHD were significantly decreased compared to healthy controls (n=22) (&gt;2.3-fold, P&lt;0.01). s and flCTLA4 expression in patients with autoGVHD was significantly decreased compared to patients without autoGVHD (&gt;2.1-fold). sCTLA4 expression in patients with alloGVHD was significantly decreased than patients without alloGVHD. Interestingly, temporal analysis revealed that the levels for sCTLA4 paralleled the recovery from GVHD implicating an active process in the establishment of non-responsiveness. CD28, CTLA4 and GITR s and fl mRNA levels in CD4+CD25+ T regulatory (Treg) cells from allo and autoBMT patients were significantly increased (7-, 41- or 22-fold, P&lt;0.01) compared to the CD4+CD25− subset. Additional studies attempted to identify the potential role of the sCTLA4 protein (encoded by the mRNA splice variant) on the regulation of the lymphocyte response mediated by Treg cells. Addition of the Treg cells to a mixed lymphocyte reaction suppressed the proliferative response of CD8+ T cells to alloantigens (75% suppression; &gt;4 fold reduction of 3H-thymidine incorporation). However, pretreatment of the Treg subset with short interfering RNA (siRNA) to knockdown sCTLA4 gene (confirmed by quantitative PCR) significantly reduced the ability of these cells to suppress the response (minimal suppression was detected, 6%). In vitro siRNA studies also indicated that Treg cells with inhibited sCTLA4 expression were unable to suppress the response of IL-2-stimulated autoreactive CD8+ T cells. Taken together, the results indicate that increased expression of CTLA4 (soluble and cell-surface associated) and the “negative” signal delivered by this molecule to the T cell may regulate the development of GVHD and help to re-establish self tolerance after BMT. Defining the role of costimulation and the modulation of this pathway on immune recognition and regulation not only provides opportunities to enhance the re-establishment of tolerance but also may help to intensify anti-tumor immunotherapeutic strategies.

scholarly journals Blockade of Interleukin 27 Signaling Attenuates Graft Versus Host Disease By Augmenting CD4+ and CD8+ Regulatory T Cell Reconstitution

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 150-150
Author(s):  
Ludovic Belle ◽  
Kimberle A. Agle ◽  
Vivian Zhou ◽  
Vanessa Yuan ◽  
Jie Sun ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Graft Versus Host Disease ◽  
Cd8 T Cells ◽  
Regulatory T Cell ◽  
Wild Type ◽  
Host Disease ◽  
Graft Versus Host ◽  
Target Organs ◽  
Ifn Γ

Abstract The interleukin-6 (IL-6) cytokine superfamily (i.e. IL-6, IL-12, and IL-23) plays a major role in the modulation of inflammatory and regulatory pathways during graft versus host disease (GVHD). IL-27, a recently discovered member of this family, is a heterodimeric cytokine that is composed of the p28 and EBI3 subunits and signals through a heterodimeric receptor composed of WSX-1 and gp130. Notably, IL-6 also uses gp130 as a signaling component which biologically links IL-27 and IL-6. IL-27 has been shown to have opposing proinflammatory and immunoregulatory effects, but its role in GVHD is not well understood. To define the functional significance of IL-27, lethally irradiated Balb/c (H-2d) mice were transplanted with C57BL/6J (H-2b) BM and spleen cells, and then treated with an anti-IL-27p28-specific antibody on days 0 and +6. p28 antibody-treated animals had significantly improved weight recovery and overall survival (47% versus 0% survival at day 60, p=0.002), as well as reduced numbers of proinflammatory CD4+ and CD8+ IFN-γ+ T cells in GVHD target organs, when compared to isotype control antibody-treated mice. A similar outcome was observed in an MHC-matched, minor antigen disparate model (B6→Balb.B), indicating that this was not a strain-specific phenomenon. Given the similarities between IL-6 and IL-27, we examined whether blockade of IL-27 promoted regulatory T cell (Treg) reconstitution as has been observed with inhibition of IL-6 signaling. Recipients transplanted with BM grafts from B6 Foxp3EGFP reporter animals and treated with p28 antibody had a significant increase in the number of CD4+ nTregs, CD4+ iTregs and CD8+ iTregs in GVHD target organs, indicating that blockade of IL-27 augmented global Treg reconstitution. In fact, inhibition of IL-27 was more effective at augmenting Treg reconstitution than comparable antibody blockade of IL-6. To further elucidate the role of IL-27, we employed transgenic IL-27−/− and IL-27R−/− animals to dissect the relevant contributions of donor and recipient populations. Paradoxically, we observed that transplantation with IL-27−/− donor grafts exacerbated GVHD mortality and augmented accumulation of proinflammatory T cells, whereas transplantation of recipient IL-27−/− mice with wild type grafts had no effect on transplant outcomes. This discordance between antibody-based and genetic studies was unexpected and led us to consider whether there were steady state alterations in T cells from IL-27−/− animals that biased these cells towards a proinflammatory phenotype. To that end, we observed that naive CD8+ T cells from IL-27−/− mice had greater IFN-γ production than wild type cells after in vitro polyclonal stimulation and CD4+ nTregs from these animals had diminished expression of CXCR3 which is critical for Treg trafficking into inflamed tissue sites. Thus, the lack of endogenous IL-27 resulted in intrinsic immune dysregulation which led to an exacerbation of GVHD after transfer of these T cells into recipients. To resolve this paradox, we employed IL-27R−/− (WSX-1−/−) mice and demonstrated that mice transplanted with IL-27R−/− grafts had enhanced weight recovery and survival providing confirmation that blockade of IL-27 signaling reduced GVHD. In addition, using IL-27R−/− Foxp3EGFP reporter mice, we observed increased frequencies and numbers of CD4+ and CD8+ Foxp3+ T cells in mice reconstituted with IL-27R−/− grafts, confirming results observed with p28 antibody blockade. Since IL-10 is a mechanism by which CD4+ Tregs suppress GVHD and IL-27 has been shown to enhance T cell-derived IL-10 secretion in nontransplant models, we examined whether IL-27 blockade adversely affected IL-10 production by Tregs. Recipients transplanted with marrow grafts from IL-10.BitFoxp3EGFP dual reporter animals and treated with p28 antibody had a significant reduction in the frequency of IL-10-producing conventional CD4+ and CD8+ T cells in GVHD target organs. Notably, however, there was no difference in the frequency of CD4+ Foxp3+ IL-10+ T cells, indicating that blockade of IL-27 signaling preferentially affected conventional T cells and had no adverse effect on CD4+ Foxp3+ T cell-derived IL-10 production. In summary, these studies demonstrate that blockade of IL-27 signaling potently augments Treg reconstitution leading to a reduction in the severity of GVHD and may therefore represent a novel strategy to reduce mortality from this disease in man. Disclosures No relevant conflicts of interest to declare.

Paradoxical effects of interleukin-18 on the severity of acute graft-versus-host disease mediated by CD4+ and CD8+ T-cell subsets after experimental allogeneic bone marrow transplantation

Blood ◽  
2004 ◽  
Vol 104 (10) ◽  
pp. 3393-3399 ◽  
Author(s):  
Chang-Ki Min ◽  
Yoshinobu Maeda ◽  
Kathleen Lowler ◽  
Chen Liu ◽  
Shawn Clouthier ◽  
...  
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Acute Gvhd ◽  
Allogeneic Bone Marrow Transplantation ◽  
Allogeneic Bone ◽  
Interleukin 18 ◽  
Graft Versus Host ◽  
Donor T Cells ◽  
Paradoxical Effects

Abstract Administration of exogenous interleukin-18 (IL-18) regulates experimental acute graft-versus-host disease (GVHD) in a Fas-dependent manner when donor CD4+ T cells are required for mortality after experimental allogeneic bone marrow transplantation (BMT). However, CD4+ and CD8+ T cells can induce acute GVHD after clinical allogeneic BMT, and the role of IL-18 in CD8+-mediated acute GVHD is unknown. We, therefore, determined the role of IL-18 in GVHD mediated by CD4+ or CD8+ T cells across major histocompatibility complex (MHC) class II- and class I-disparate allogeneic BMT, respectively. Administering IL-18 significantly increased survival in CD4+-mediated GVHD but reduced survival in CD8+-mediated GVHD. This increase in deaths was associated with significantly greater clinical, biochemical, and histopathologic parameters of GVHD damage and was independent of Fas expression on donor T cells. Administering IL-18 significantly enhanced allospecific cytotoxic function and expansion of CD8+ cells. Endogenous IL-18 was critical to GVHD mediated by CD8+ donor T cells because IL-18 receptor-deficient donors caused significantly less GVHD but exacerbated CD4+-mediated, GVHD-related death. Furthermore, administering anti-IL-18 monoclonal antibody significantly reduced CD8+-mediated, GVHD-related death. Together these findings demonstrate that IL-18 has paradoxical effects on CD4+ and CD8+ cell-mediated GVHD. (Blood. 2004;104:3393-3399)

scholarly journals Bone Marrow NK1.1− and NK1.1+ T Cells Reciprocally Regulate Acute Graft versus Host Disease

1999 ◽  
Vol 189 (7) ◽  
pp. 1073-1081 ◽  
Author(s):  
Defu Zeng ◽  
David Lewis ◽  
Sussan Dejbakhsh-Jones ◽  
Fengshuo Lan ◽  
Marcos García-Ojeda ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Graft Versus Host Disease ◽  
Cd8 T Cells ◽  
Peripheral Blood ◽  
T Cell Subsets ◽  
Host Disease ◽  
Graft Versus Host ◽  
Ifn Γ

Sorted CD4+ and CD8+ T cells from the peripheral blood or bone marrow of donor C57BL/6 (H-2b) mice were tested for their capacity to induce graft-versus-host disease (GVHD) by injecting the cells, along with stringently T cell–depleted donor marrow cells, into lethally irradiated BALB/c (H-2d) host mice. The peripheral blood T cells were at least 30 times more potent than the marrow T cells in inducing lethal GVHD. As NK1.1+ T cells represented &lt;1% of all T cells in the blood and ∼30% of T cells in the marrow, the capacity of sorted marrow NK1.1− CD4+ and CD8+ T cells to induce GVHD was tested. The latter cells had markedly increased potency, and adding back marrow NK1.1+ T cells suppressed GVHD. The marrow NK1.1+ T cells secreted high levels of both interferon γ (IFN-γ) and interleukin 4 (IL-4), and the NK1.1− T cells secreted high levels of IFN-γ with little IL-4. Marrow NK1.1+ T cells obtained from IL-4−/− rather than wild-type C57BL/6 donors not only failed to prevent GVHD but actually increased its severity. Together, these results demonstrate that GVHD is reciprocally regulated by the NK1.1− and NK1.1+ T cell subsets via their differential production of cytokines.

JAK1/2 Inhibition Suppresses T Cell Proliferation and Inflammatory Cytokine Production In An Allogeneic Setting and Ameliorates Graft-Versus-Host Disease (GvHD) In a Murine GvHD Model

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 3253-3253
Author(s):  
Silvia Spoerl ◽  
Sophia Chen ◽  
Michael Bscheider ◽  
Nimitha Mathew ◽  
Martina Schmickl ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Cd8 T Cells ◽  
Inflammatory Cytokine ◽  
Cytokine Production ◽  
Ifn Gamma ◽  
Inflammatory Cytokine Production ◽  
Graft Versus Host ◽  
Cell Responses

Abstract Graft-versus-host-disease (GvHD) constitutes a severe complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). In GvHD, tissue damage is mediated by pro-inflammatory cytokines. Cytokine responses are mediated by activated Janus kinases (JAKs). We hypothesized that JAK1/2 inhibition might reduce T effector cell responses and inflammatory cytokine production in an allogeneic system, thereby ameliorating GvHD. We established an allogeneic (major mismatch) cell culture system using naive BALB/c CD4+ CD62Lhigh T cells that were co-cultured with C57BL/6 (B6) bone marrow derived dendritic cells (DC). JAK 1/2 signaling was specifically blocked using the small molecule inhibitor Ruxolitinib (INCB018424). By using a 3H Thymidine proliferation assay, we found that Ruxolitinib was able to inhibit the proliferation of allogeneic CD4+ effector T cells. In our intracellular cytokine staining experiments Ruxolitinib was able to impair the differentiation of naïve T cells into IFN-gamma and IL17A-producing T effector cells. Both cytokines – IFN-gamma and IL-17A – have been linked to aggravated courses of GvHD severity. In vivo administration of Ruxolitinib signficantly reduced lethal GvHD after allo-HSCT in a murine major mismatch model. BALB/c recipient mice were irradiated and received C57BL/6 (B6) T cell depleted bone marrow along with CD4+ and CD8+ T cells. Animals that were treated with an oral gavage of Ruxolitinib twice daily displayed significantly lower serum levels of IFN-gamma, TNF-alpha, IL-10 and IL-12p70, lower histological and clinical GvHD scores and improved overall survival compared to the control group. By using luciferase-positive (luc+) CD4+ and CD8+ T cells, we were able to visualize and quantify T cell migration after GvHD induction. The recipient mice received luc+ T cells along with conventional T cell depleted bone marrow. We were able to detect the luc+ T cell signal with a bioluminescence imaging system. At day 9 after allogeneic bone marrow transplantation, migration of donor T cells to GvHD target sites was significantly reduced in Ruxolitinib treated mice compared to control mice. In summary, our data demonstrate that suppression of inflammatory cytokine responses by JAK1/2 inhibitors modulates T effector cell responses in an allogeneic setting and improves survival in a murine major mismatch GvHD model. These observations suggest that JAK1/2 inhibition might be used for the treatment of GvHD following allo-HSCT. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Prevention of Acute Graft-Versus-Host Disease Using an Engineered Mouse Orthogonal IL-2/IL-2Rβ Regulatory T Cells

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 1688-1688
Author(s):  
Teresa Lopes Ramos ◽  
Sara B. Wagers ◽  
Po-Yu Lin ◽  
Toshihito Hirai ◽  
Leon Su ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Board Of Directors ◽  
Graft Versus Host Disease ◽  
Research Funding ◽  
Acute Gvhd ◽  
Treg Cells ◽  
Advisory Committees ◽  
Graft Versus Host ◽  
Acute Graft

Abstract Regulatory T cells are a critical population for the maintenance of self-tolerance and the use of these cells for therapeutic purpose in transplantation has emerged based on their capacity to control immune activation, exhibiting positive therapeutic potential to ameliorate acute graft-versus-host disease (GvHD), one of the major complications after allogeneic bone marrow transplantation (allo-BMT). One of the remaining technical challenges in the clinical practice of Treg administration is the number of Treg required to effectively prevent GvHD. In this study, we engineered murine Treg cells to express an altered IL-2 receptor β subunit that selectively interacts with an orthogonal mutant IL-2 (oIL2) without interacting with the natural cytokine or receptor counterparts, thus preventing acute -GvHD. For investigation of the suppressive function of Treg orthogonal IL-2/IL-2 receptor system in a murine major mismatch of acute GvHD model, the BALB/c mice were lethally irradiated (8.8 Gy) and transplanted with 5x10 6 T cell-depleted bone marrow cells (TCD-BM) from C57Bl/6 mice alone or together with fresh 1x10 6 C57Bl/6 FoxP3 DRT-GFP-luc+ Treg cells (fTreg) or with 0.2x10 6 and 1x10 6 oTreg (transduced with the construct for murine oIL-2 receptor β chain) on day 0. On day 2, C57Bl/6 Tcon cells (1x10 6) were injected to induce GvHD. Recipients were treated with PBS or oIL-2 (25K IU/daily) for 14 days and then 3 times a week for another 14 days. Using bioluminescence imaging, we observed that ortho IL2Rβ +Treg not only maintains migration ability to the gut, mLN and LN but also exhibited significant expansion ability treated with oIL2 at day 7 post-transplant. The expansion of oTreg with PBS and fTreg was observed on day 14 after allo-BMT. The administration of oTreg-oIL2 and fTreg (1:1 Tcon:Treg ratio) significantly increased the survival of the mice compared to Tcon group (P&lt;0.0001 and P=0.0001, respectively). Furthermore, the group oTreg-oIL2 (1:1) showed a significant increase in the survival curve compared to the fTreg group (P=0.03). When decreasing the number of oTreg administrated (5:1 Tcon:Treg ratio) a significant increase in the survival (Tcon vs Tcon:oTreg (5:1)+oIL2; P&lt;0.0001) in the oIL2 group. Similar survival curves were observed between this group and fTreg group (1:1). oIL-2 protein injection was able to selectively expand the oTreg in vivo, with a significantly increased percentage of Foxp3 GFP+ in CD4 + T cells (PBS;14.96±2.4%, oIL-2; 27.76±23.6%, P=0.02) in the peripheral blood (PB) and spleen after 7 days post-transplant. We also observed a significant decrease in the percentage of effector T cells in the LN, mLN and spleen of the mice transplanted with oTreg and oIL-2 administration after 7 days post transplantation compared with GvHD group. We further demonstrated that ortho-IL2 administration-maintained graft-versus-leukemia (GvL) responses against lymphoma cells (A20). Engineered orthogonal cytokine receptor improves the suppressing abilities of Treg potential in a murine model of acute GvHD with the ability to maintain GvL, representing a novel approach to the utilization of Treg in Allogeneic Hematopoietic Stem Cell Transplant. Disclosures Garcia: Synthekine: Membership on an entity's Board of Directors or advisory committees. Blazar: Tmunity Therapeutics: Other: Co-founder; Equilibre Pharmaceuticals Corp: Research Funding; Carisma Therapeutics, Inc: Research Funding; Rheos Medicines: Research Funding; BlueRock Therapeutics: Membership on an entity's Board of Directors or advisory committees, Research Funding; Magenta Therapeutics: Membership on an entity's Board of Directors or advisory committees.

scholarly journals Rapamycin and IL-2 reduce lethal acute graft-versus-host disease associated with increased expansion of donor type CD4+CD25+Foxp3+ regulatory T cells

Blood ◽  
2011 ◽  
Vol 118 (8) ◽  
pp. 2342-2350 ◽  
Author(s):  
Ho-Jin Shin ◽  
Jeanette Baker ◽  
Dennis B. Leveson-Gower ◽  
Aaron T. Smith ◽  
Emanuela I. Sega ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Acute Gvhd ◽  
Graft Versus Host ◽  
Donor Type ◽  
Ifn Γ ◽  
And Function ◽  
Acute Graft ◽  
Stimulation Of

Abstract Previous work has demonstrated that both rapamycin (RAPA) and IL-2 enhance CD4+CD25+Foxp3+ regulatory T-cell (Treg) proliferation and function in vitro. We investigated whether the combination of RAPA plus IL-2 could impact acute GVHD induction after bone marrow transplantation (BMT). RAPA plus IL-2 resulted in improved survival and a reduction in acute GVHD lethality associated with an increased expansion of donor type CD4+Foxp3+ Tregs and reduced CD4+CD25− conventional T cells (Tcons). RAPA plus IL-2, but not either drug alone, increased both expansion of donor natural Tregs and conversion of induced Tregs from donor CD25− Tcons while IL-2 alone increased conversion of Tregs from CD25− Tcon. RAPA plus IL-2 treatment resulted in less production of IFN-γ and TNF, cytokines known to be important in the initiation of acute GVHD. These studies indicate that the pharmacologic stimulation of T cells with IL-2 and the suppression of Tcon proliferation with RAPA result in a selective expansion of functional Tregs and suppression of acute GVHD.

Critical Role of the Th1 Transcription Factor T-Bet in An Animal Model of Immune-Mediated Bone Marrow Failure

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1037-1037
Author(s):  
Yong Tang ◽  
Jichun Chen ◽  
Neal Young
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Cd8 T Cells ◽  
Bone Marrow Failure ◽  
Critical Role ◽  
Wild Type ◽  
Immune Mediated ◽  
Ifn Γ ◽  
Th1 Immune Response

Abstract Patients with aplastic anemia have elevated T-bet, a Th1 transcription factor, in peripheral blood CD4 and CD8 T cells, suggesting that T-bet over-expression and dysregulated Th1 immune response contributes to pathophysiology of marrow failure (Solomou EE et al., Blood. 2006; 107:3983). In the present study, we studied the role of T-bet in inducing bone marrow failure in a mouse model of immune-mediated BM failure, employing mice engineered with a germline T-bet deletion as lymphocyte donors. Compared with T-bet +/+ wild-type controls, T-bet−/− mice have similar cellular composition in various lymphohematopietic tissues including peripheral blood, spleen, thymus, lymph nodes (LN), and BM. Incubation of effector T-bet−/− LN cells with MHC-mismatched target CByB6F1 (F1) BM cells in an in vitro cytotoxicity assay resulted in a significantly lower proportion of apoptotic target cells than did wild-type T-bet+/+ LN effector cells, suggesting that T-bet−/− effector LN cells are functionally defective. While infusion of 5×106 wild-type T-bet+/+ LN cells into sublethally-irradiated F1 mice led to severe pancytopenia and aplastic bone marrow in recipient mice, infusion of the same number of T-bet−/− LN cells failed to result in marrow failure, and recipients had relatively normal blood counts and bone marrow cellularity. By flow cytometry, both expansion of CD4+ and CD8+ T cells and elevation in intracellular Th1 cytokine gamma interferon (IFN-γ), which are characteristic of marrow cells in recipients received B6 LN cells, were absent in recipients receiving T-bet −/− LN cells. Serum IFN-γ concentration in F1 mice infused with T-bet −/− LN cells was similar to the level in F1 control mice received TBI alone, and both were significantly lower than serum IFN-γ in recipients of wild-type B6 LN cells. In contrast, serum TGF-γ concentration was higher in F1 mice that received TBI alone or TBI plus T-bet −/− LN cell infusion, compared with mice that received TBI plus B6 LN cells. An increase of T-bet −/− LN cell infusion to 10×106 cells per recipient led to very mild BM failure. Contrary to the markedly increased number of CD4+ and CD8+ T cells and elevated IFN-γ level in the BM of F1 mice which have received wild type B6 LN cells, F1 mice infused with T-bet −/− LN have low CD4+ and CD8+ cells and low IFN-γ level in the BM similar to F1 mice received TBI alone, but they show increased IL4 and IL17 levels within bone marrow T cells, indicating that the diminished Th1 immune response due to T-bet deficiency was partially compensated by up-regulated Th2 and Th17 responses. Our data demonstrated that T-bet plays a critical role in immune mediated bone marrow failure. Approaches targeting to T-bet signal pathway may lead to novel treatment for bone marrow failure and other autoimmune diseases.

Inducible Costimulator Gene Transduced Bone Marrow Derived Mesenchymal Stem Cells Attenuate the Severity of Acute Graft-Versus-Host Disease in a Mouse Haploidentical Model.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2996-2996
Author(s):  
Dan Yang ◽  
Jian-Min Wang ◽  
Li-ping Wang ◽  
Hong Zhou
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
T Cells ◽  
Mesenchymal Stem Cells ◽  
Cd4 T Cells ◽  
Acute Gvhd ◽  
Early Stage ◽  
Graft Versus Host ◽  
Ifn Γ ◽  
Acute Graft

Abstract Abstract 2996 It is reported that both block of costimulator (ICOS) or its ligand signal pathway and transfusion of mesenchymal stem cells (MSCs) can reduce the severity of acute graft-versus-host disease (GvHD). Using a MHC-haploidentical murine model of acute GvHD, we investigated the effects of MSC-ICOS-EGFP, bone marrow derived MSCs transduced with ICOS-gene using an adenoviral vector, on acute GvHD. Lethally irradiated CB6F1 mice were transplanted with bone marrow cells and spleen cells from C57BL/6 mice, and injected with MSC-ICOS-EGFP, MSCs, ICOS-Ig fusion protein or the diluent (control), respectively. Survival in recipients of MSC-ICOS-EGFP was significantly higher than that in GVHD group (74.29±7.39% vs 0, P=0.000, n=35), and also higher than in MSCs, ICOS-Ig, or MSCs+ICOS-Ig groups (74.29±7.39% vs 42.86±8.36% or 48.57±8.45% or 50.43±8.45%, P=0.0039 or 0.0323 or 0.0418, n=35). The lower mortality in recipients of MSC-ICOS-EGFP was related to lower incidence of acute GvHD. MSC-ICOS-EGFP can reduce the number of CD4+T cells in the early stage of GvHD due to the increased apoptosis, and it can produce CD4+CD25+Treg cells meanwhile. The effect on GvHD in recipient of MSC-ICOS-EGFP was also associated with higher serum levels of IL-4, IL-10 and lower levels of IFN-γ, IL-2, IL-12, IL-17A, as well as inducing expression of GATA-3, STAT6 transcription factors while inhibiting expression of T-bet, STAT4, ROR-rt in spleen. In vitro, the CD4+T cells of donor were sharply inhibited by MSC-ICOS-EGFP and the B7h blockade. In MLRs we can also detect lower levels of IFN-γ, IL-2and higher levels of IL-4, IL-10. These data indicate that MSC-ICOS-EGFP is a more prospective strategy for acute GvHD prevention, which had a synergism effect between MSCs and ICOS-Ig. The mechanisms of MSC-ICOS-EGFP on acute GvHD were closely associated with modulation of CD4+T cells, CD4+CD25+Tregs, and regulation of the Th1/Th2/Th17 balance in the early stage of GvHD. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Signaling Through the Type 2 Cannabinoid Receptor Regulates the Severity of Acute and Chronic Graft versus Host Disease

Blood ◽  
2020 ◽  
Author(s):  
Cheng Yin Yuan ◽  
Vivian Zhou ◽  
Garrett Sauber ◽  
Todd M Stollenwerk ◽  
Richard Komorowski ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Graft Versus Host Disease ◽  
Cd8 T Cells ◽  
Acute Gvhd ◽  
Cannabinoid Receptor ◽  
Therapeutic Targeting ◽  
Host Disease ◽  
Graft Versus Host

Graft versus host disease (GVHD) pathophysiology is a complex interplay between cells that comprise the adaptive and innate arms of the immune system. Effective prophylactic strategies are therefore contingent upon approaches that address contributions from both immune cell compartments. In the current study, we examined the role of the type 2 cannabinoid receptor (CB2R) which is expressed on nearly all immune cells and demonstrated that absence of the CB2R on donor CD4+ or CD8+ T cells, or administration of a selective CB2R pharmacological antagonist, exacerbated acute GVHD lethality. This was accompanied primarily by the expansion of proinflammatory CD8+ T cells indicating that constitutive CB2R expression on T cells preferentially regulated CD8+ T cell alloreactivity. Using a novel CB2R-EGFP reporter mouse, we observed significant loss of CB2R expression on T cells, but not macrophages, during acute GVHD, indicative of differential alterations in receptor expression under inflammatory conditions. Therapeutic targeting of the CB2R with the agonists, tetrahydrocannabinol (THC) and JWH-133, revealed that only THC mitigated lethal T cell-mediated acute GVHD. Conversely, only JWH-133 was effective in a sclerodermatous chronic GVHD model where macrophages contribute to disease biology. In vitro, both THC and JWH-133 induced arrestin recruitment and ERK phosphorylation via CB2R, but THC had no effect on CB2R-mediated inhibition of adenylyl cyclase. These studies demonstrate that the CB2R plays a critical role in the regulation of GVHD and suggest that effective therapeutic targeting is dependent upon agonist signaling characteristics and receptor selectivity in conjunction with the composition of pathogenic immune effector cells.

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