Graft-Versus-Host Tolerance in Dogs Following T-Cell Depleted Bone Marrow Transplantation with Absorbed ATG or CD6-Antibody.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2338-2338
Author(s):  
Julia Zorn ◽  
Hans Jochem Kolb
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Stem Cell ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Nk Cells ◽  
Marrow Transplantation ◽  
Graft Versus Host ◽  
Host Tolerance ◽  
Rabbit Complement

Abstract Graft-versus-host disease (GvHD) is the major obstacle of allogeneic stem cell transplantation. Depletion of T-cells from the graft reduces the risk of GvHD, but results in a higher risk of leukemia relapse. Adoptive immunotherapy with donor lymphocyte transfusion (DLT) has been shown to control leukemia in patients after T-cell depleted allogeneic stem cell transplantation. However, GvHD may occur, if DLT is given too early after transplantation. In canine models of DLA-identical and DLA-haploidentical bone marrow transplantation, we compared different methods of T-cell depletion (TCD) and investigated the potential of DLT at different times after transplantation to induce GvHD. T-cell depletion was performed either with absorbed anti-thymocyte globuline (aATG) or with a combination of CD6-antibody and baby rabbit complement. ATG was absorbed with erythrocytes, liver, kidney and spleen for eliminating antibodies against stem cells. CD6-antibody (M-T606) and rabbit complement depleted T-cells effectively without affecting hematopoietic progenitor cells. Unlike aATG, monoclonal CD6-antibody spares natural killer (NK) cells and some CD8-positive cells. Treatment of bone marrow with aATG prevented GvHD in 9 dogs following DLA-identical transplantation. DLT on days 1 and 2 or 21 and 22 induced fatal GvHD in two dogs each. However, it did not induce GvHD when given on days 61 and 62 and later. In DLA-haploidentical bone marrow recipients, non-manipulated marrow produced fatal GvHD in all dogs (n=7), whereas marrow treated with aATG (vol:vol 1:100 and 1:200) produced fatal GvHD in 5 out of 16 dogs only. CD6-depletion prevented GvHD in 3 of 3 DLA-haploidentically transplanted dogs. DLT produced fatal GvHD in one dog each, when given on day 3, 7 or 14 after CD6-depleted haploidentical bone marrow transplantation. However, it produced fatal GvHD in only 2 of 4 dogs transfused on day 20 post grafting. Thus, DLT could be given earlier in DLA-haploidentical animals transplanted with CD6-depleted marrow than in DLA-identical animals transplanted with aATG treated marrow without producing GvHD. These findings support the hypothesis that graft-versus-host tolerance can be induced earlier with grafts not depleted of NK cells. NK cells in the graft may inactivate host dendritic cells necessary for the induction of GvHD. In grafts depleted with aATG, NK cells are depleted as well, because aATG still retains broad specificity despite extensive absorptions. This leaves host DCs unaffected. Transfused donor T-cells encountering this environment will thus be activated which results in severe GvHD. In contrast, monoclonal CD6-antibody spares NK cells, so that donor lymphocytes cannot be activated by host DCs at the time of DLT and thus won’t trigger GvHD. CD6-depletion is the preferred method if adoptive immunotherapy with DLT is planned.

scholarly journals Extrathymic T Cell Deletion and Allogeneic Stem Cell Engraftment Induced with Costimulatory Blockade Is Followed by Central T Cell Tolerance

1998 ◽  
Vol 187 (12) ◽  
pp. 2037-2044 ◽  
Author(s):  
Thomas Wekerle ◽  
Mohamed H. Sayegh ◽  
Joshua Hill ◽  
Yong Zhao ◽  
Anil Chandraker ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Stem Cell ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Marrow Transplantation ◽  
T Cell Tolerance ◽  
Cell Tolerance ◽  
Costimulatory Blockade ◽  
Cell Engraftment

A reliable, nontoxic method of inducing transplantation tolerance is needed to overcome the problems of chronic organ graft rejection and immunosuppression-related toxicity. Treatment of mice with single injections of an anti-CD40 ligand antibody and CTLA4Ig, a low dose (3 Gy) of whole body irradiation, plus fully major histocompatibility complex–mismatched allogeneic bone marrow transplantation (BMT) reliably induced high levels (>40%) of stable (>8 mo) multilineage donor hematopoiesis. Chimeric mice permanently accepted donor skin grafts (>100 d), and rapidly rejected third party grafts. Progressive deletion of donor-reactive host T cells occurred among peripheral CD4+ lymphocytes, beginning as early as 1 wk after bone marrow transplantation. Early deletion of peripheral donor-reactive host CD4 cells also occurred in thymectomized, similarly treated marrow recipients, demonstrating a role for peripheral clonal deletion of donor-reactive T cells after allogeneic BMT in the presence of costimulatory blockade. Central intrathymic deletion of newly developing T cells ensued after donor stem cell engraftment had occurred. Thus, we have shown that high levels of chimerism and systemic T cell tolerance can be reliably achieved without myeloablation or T cell depletion of the host. Chronic immunosuppression and rejection are avoided with this powerful, nontoxic approach to inducing tolerance.

Bone Marrow T Cells Are More Effective Than Peripheral T Cells in Inducing Chimeric Conversion Following MHC-Mismatched Nonmyeloablative Bone Marrow Transplantation

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4593-4593
Author(s):  
Seok-Goo Cho ◽  
Hyunsil Park ◽  
Min Jung Park ◽  
Ho-Youn Kim ◽  
Jong-Wook Lee ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Marrow Transplantation ◽  
Memory T Cells ◽  
Mixed Chimerism ◽  
Graft Versus Host ◽  
Peripheral T Cells ◽  
Mixed Chimeras

Abstract Background & Objectives: Recently, T cells in BM have attracted renewed interest because they are now known to have different surface phenotypes, subsets, and activation states from those in the periphery. Memory T cells undergo extensive migration from the blood to the BM and vice versa. The BM plays an important role in preferential homing and extensive proliferation of memory T cells, and contributes considerably to the longlived memory T cell pool. BM T cells are more activated than their splenic counterparts and have a higher rate of local proliferation. Although BM-T (NK1.1– CD4+ or CD8+) cells did not induce lethal GVH disease, even at high cell numbers, BM-T cells mediated vigorous graft-versus-tumor activity and facilitated engraftment of hematopoietic progenitor cells. These studies suggested that BM-T cells could be a useful cellular source for adoptive immunotherapy following ABMT, instead of peripheral T cells. Non-myeloablative bone marrow transplantation (NMT) and allogeneic mixed chimerism can provide an environment adequate for diminishing susceptibility to DLI-mediated GVHD and an immunological platform for DLI in both mouse and human models. In patients treated with DLI, a successful GVL effect is often associated with conversion to complete donor chimerism, supporting the concept of a graft-versus-host (GVH) response as part of the GVL effect. Thus, a quiet chimeric conversion following DLI is desirable to reach an optimal DLI-mediated GVL effect, without the occurrence of GVHD. Although in a mouse model, the administration of non-tolerant donor spleen cells to established mixed chimeras has been shown to convert mixed hematopoietic chimerism to full donor chimerism, without the concomitant development of GVHD, DLI in humans frequently results in serious GVHD and life-threatening complications. However, the use of BM-T cells, as compared with spleen T cells (SP-T), as the DLI source has not been investigated in allogeneic mixed chimerism prepared with NMT. In this study, we evaluated the beneficial alloreactivity of DLI using cryopreserved BM-T cells, a by-product obtained during the T cell depletion (TCD) procedure in BM grafting, to effectively induce chimeric conversion without the occurrence of GVHD in MHC-mismatched NMT. Methods: Cells were prepared using established procedures. During the T cell depletion (TCD) procedure in BM grafting, BM-T cells were obtained as a by-product and then cryopreserved for subsequent DLI using BM-T cells 21 days after the bone marrow transplant. Results: The administration of 5–10 × 105 BM-T (Thy1.2+) cells in mixed chimeras resulted in complete chimeric conversion, with self-limited graft-versus-host disease (GVHD) and no pathological changes. However, the administration of 5–10 × 105 SP-T (Thy1.2+) cells resulted in persistent mixed chimerism, with pathological GVHD signs in the liver and intestine. Conclusion: Our results suggest that DLI using BM-T cells, even in small numbers, could be more potent for inducing chimeric conversion in mixed chimerism than DLI using SP-T cells. Further study is needed to determine whether cryopreserved BM-T cells are an effective cell source for DLI to consolidate donor-dominant chimerism in clinical practice, without concerns about GVHD.

scholarly journals Lethal graft-versus-host disease after bone marrow transplantation across minor histocompatibility barriers in mice. Prevention by removing mature T cells from marrow.

1978 ◽  
Vol 148 (6) ◽  
pp. 1687-1698 ◽  
Author(s):  
R Korngold ◽  
J Sprent
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Graft Versus Host Disease ◽  
Marrow Transplantation ◽  
Host Disease ◽  
Graft Versus Host ◽  
High Incidence ◽  
Marrow Cells

In two situations, transfer of normal unsensitized bone marrow cells into heavily irradiated H-2-identical allogeneic mice caused a high incidence of lethal chronic graft-versus-host disease (GVHD), i.e. mortality occuring between days of 20 and 80 postirradiation. Minor histocompatibility determinants appeared to be the main target for eliciting GVHD. Removing mature T cells from the marrow with anti-Thy 1.2 serum and complement before injection prevented GVHD. On the basis of adding purified T cells to T-cell-depleted marrow cells, it was concluded that contamination of the marrow with as few as 0.3% T cells was sufficient to cause a high incidence of lethal GVHD in certain situations. No GVHD was found with the injection of non-T cells (Thy 1.2-negative cells) or with tolerant T cells. Irradiated recipients of T-cell-depleted marrow cells remained in good health for prolonged periods. These mice showed extensive chimerism with respect to the donor marrow, normal numbers of T and B cells and were immunocompetent. The data provide no support for the view that chronic GVHD developing after bone marrow transplantation in man is the result of an attack by the progeny of the donor stem cells. The results imply that mature T cells contaminating marrow inocula are probably the main cause of GVHD seen in the clinical situation.

scholarly journals Early CD30 Signaling Is Critical for Adoptively Transferred CD4+CD25+ Regulatory T Cells in Prevention of Acute Graft Versus Host Disease.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3176-3176
Author(s):  
Robert Zeiser ◽  
Vu H. Nguyen ◽  
Jing-Zhou Hou ◽  
Andreas Beilhack ◽  
Elizabeth A. Zambricki ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Regulatory T Cells ◽  
Graft Versus Host Disease ◽  
Marrow Transplantation ◽  
Host Disease ◽  
Graft Versus Host ◽  
Acute Graft

Abstract CD4+CD25+ regulatory T cells (Treg) have been demonstrated to reduce the severity of acute graft-versus-host disease (aGvHD) in murine models of bone marrow transplantation. However, the surface molecules that are critical for suppression are unclear. The TNF-R superfamily member CD30 has been shown to be expressed on regulatory T cells that down-modulate nickel specific immune responses and to be relevant for Treg mediated protection from allograft rejection. Deficiency of the CD30 molecule (CD30−/−) is associated with impaired thymic negative selection and augmented T cell autoreactivity. Therefore, we investigated the role of CD30 signaling in Treg function in an aGvHD model. Treg derived from CD30−/ − animals were significantly less effective in preventing aGvHD lethality (wt vs. CD30−/ − p=0.002). Signal intensity derived from expanding luciferase expressing alloreactive conventional T cells (Tconv) was significantly higher if CD30−/ − Treg as compared to wt Treg (p=0.007) were transferred as assessed by bioluminescencent based imaging. Blockade of the CD30/CD153 pathway with a neutralizing anti-CD153 mAb during the early (days −2 to +4) but not late (days +4 to +10) phase of adoptive Treg transfer reduced Treg mediated protection from proinflammatory cytokine accumulation and apoptosis of donor-type CD4 and CD8 T cells. In vivo bioluminescence imaging demonstrated intact Treg homing, but reduced expansion when CD153 was blocked during the early phase after adoptive transfer. CD30 surface expression on Treg increased with alloantigen exposure and CD153 expression on recipient-type dendritic cells increased in the presence of an irradiation induced proinflammatory environment but not when T cell depleted bone marrow and Tconv were transferred into non-irradiated Rag 2−/ −γc−/ − recipients. These data are the first to demonstrate that early CD30 signaling is relevant for Treg mediated aGvHD protection after major MHC mismatch bone marrow transplantation.

scholarly journals Human HLA-specific T-cell clones with stable expression of a suicide gene: a possible tool to drive and control a graft-versus-host- graft- versus-leukemia reaction?

Blood ◽  
1996 ◽  
Vol 88 (3) ◽  
pp. 1098-1103
Author(s):  
G Gallot ◽  
MM Hallet ◽  
J Gaschet ◽  
JF Moreau ◽  
R Vivien ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Marrow Transplantation ◽  
Host Recognition ◽  
T Cell Clones ◽  
Graft Versus Host ◽  
Graft Versus Leukemia ◽  
Cell Clones

Allogeneic bone marrow transplantation is still limited by the morbidity and mortality caused by graft-versus-host disease (GVHD), resulting from host recognition by donor T lymphocytes. It is possible to drastically reduce the T-cell content of the graft. However, transplanted T cells can also have a beneficial effect by graft enhancement and the graft-versus-leukemia effect. How can we keep the beneficial GVL effect while protecting the patient from possible GVHD? A recent report proposed the ex vivo transfer of the herpes simplex thymidine kinase (HSv-tk) gene into donor T cells before their infusion with hematopoietic stem cells. This procedure is expected to allow selective donor T-cell depletion with ganciclovir should GVHD occur, but it has two major drawbacks: reinjection of a fraction of untransfected T cells cannot be avoided and heterogeneity of the transfected population results in increased risks such as HSv-tk gene instability or dysfunction of some of the transfected T cell. Alternative approaches must be considered. We demonstrate here the feasibility of generating HSv-tk transfected HLA-specific CD4+ cytotoxic T-cell clonal populations, in which 100% of the cells have the HSv-tk gene inserted at a single site within their genome. These clones retained their specificity, their function, and their sensitivity to ganciclovir treatment. Our approach is not limited to bone marrow transplantation. Indeed, this procedure represents a useful alternative to retroviral gene transduction and is applicable to every circumstance where clinical use of gene modified T-cell clones is to be considered.

scholarly journals Delayed infusion of immunocompetent donor cells after bone marrow transplantation breaks graft-host tolerance allows for persistent antileukemic reactivity without severe graft-versus-host disease

Blood ◽  
1995 ◽  
Vol 85 (11) ◽  
pp. 3302-3312 ◽  
Author(s):  
BD Johnson ◽  
RL Truitt
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Spleen Cells ◽  
Graft Versus Host ◽  
Donor Cells ◽  
Donor T Cells ◽  
Antileukemic Effect ◽  
Host Tolerance

The development of graft-host tolerance after bone marrow transplantation (BMT) is crucial to avoid the problems of graft-versus-host disease (GVHD) and graft rejection. GVHD can be eliminated by depleting mature donor T cells from the BM inoculum, thereby facilitating the development of graft-host tolerance. However, T-cell depletion often results in an increased incidence of graft rejection and an increased frequency of leukemia relapse. Thus, although graft-host tolerance is a desirable outcome, it can pose a significant threat to leukemia-bearing hosts. Using a major histocompatability complex (MHC)-matched allogeneic model of BMT (B10.BR into AKR), we found that irradiated recipients given donor BM alone displayed mixed T-cell chimerism and did not develop GVHD. Graft-host tolerance developed by 8 weeks after BMT in these chimeras, and they were susceptible to low-dose leukemia challenge. When sufficient numbers of donor spleen cells, as a source of T-cells, were added to the BM graft, AKR hosts developed severe and lethal GVHD. Antihost reactive donor T cells persisted in chimeras undergoing GVHD, indicating that graft-host tolerance did not develop. When administration of the spleen cells was delayed for 7 to 21 days after BMT, there was significantly less mortality because of GVHD. Day 21 was the optimal time for infusion of cells without development of GVHD. Graft-host tolerance was broken by the delayed infusion of donor cells, as indicated by the persistence of antihost-reactive donor T cells in these chimeras in T-cell receptor cross-linking and mixed lymphocyte reaction assays. Importantly, the persistence of antihost-reactive donor T cells correlated with along-term antileukemic effect that was still present at 100 days after transplant. Multiple infusions of immunocompetent donor cells could be administered without increasing the risk for GVHD if delayed until 21 days post-BMT. Delayed infusions of donor spleen cells also resulted in a long-term antileukemic effect in the absence of GVHD in an MHC-haplotype-mismatched model of BMT (SJL into [SJL x AKR]F1). Although delayed infusion of normal donor cells did not induce GVHD, spleen cells from donors previously sensitized to host alloantigens induced GVHD when infused 21 days after BMT. Thus, the ability of previously activated cells to induce GVHD was not inhibited in the same manner as naive cells. Results from limiting dilution analysis assays indicated that alloactivated interleukin-2-secreting CD4+ T cells were preferentially inhibited over cytolytic T cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Promotion of Stem Cell-Derived New T Cell Generation by CD62L− Memory T Cells Requires Alloantigen Recognition.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3041-3041
Author(s):  
Benny J. Chen ◽  
Xiuyu Cui ◽  
Jessica Son ◽  
Nelson J. Chao
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Host Resistance ◽  
Marrow Transplantation ◽  
Cell Generation ◽  
Promoting Effect ◽  
Graft Versus Host ◽  
Functional Relevance

Abstract We recently demonstrated that non-GVHD-inducing CD62L− memory T cells promote T cell regeneration from hematopoietic stem/progenitor cells in the C57BL/6 (H2b) to BALB/c (H2d) bone marrow transplantation model (BLOOD 2004,103:1534). In this model, 1×107 T cell depleted bone marrow from C57BL/6, CD45.2, Thy1.2 mice and 1×106 CD62L− T cells from C57BL/6, CD45.1, Thy1.1 mice are used and the recipient mice are lethally irradiated. This animal model allows us to differentiate the T cells of different origin by flow cytometry. In the current study, we further investigated how CD62L− T cells promote new T cell generation and whether there is any functional relevance using the same model. Previous data suggest that the promoting effect of CD62L− T cells on stem cell-derived new T cell generation is associated with the facilitation of engraftment because host radioresisdant T cells were depleted in CD62L− T cell recipients but not in the T cell-depleted bone marrow control mice. To determine whether CD62L− T cells promote new T cell generation by overcoming T cell-mediated host resistance, we performed the experiment using SCID mice as recipients. SCID mice lack both T and B cells. In consistent with the previous results, serial peripheral blood T cell counts following bone marrow transplantation demonstrated that the promoting effect of CD62L− T cells were dramatically decreased in the SCID recipients, indicating that CD62L− T cell promote new T cell regeneration by overcoming host resistance. To examine whether CD62L− T cells overcome host resistance by “veto effect”, we performed the experiment using CD62L− T cells from (BALB/cxC57BL/6)F1 mice. T cells from F1 mice do not recognize the parental recipient as non-self and can not initiate classical T cell response against the parental antigens. It was previously reported by other investigators that T cells from F1 mice preserve “veto effect”. However, no promoting effect was observed in the recipients of CD62L− T cells from the F1 mice, demonstrating that “veto effect” is not involved. Rather, alloantigen recognition by donor CD62L− T cells is required for the promoting effect. In agreement with the hypothesis that CD62L− T cells promote new T cell generation by enhancing engraftment through overcoming host resistance, both white cell and platelet recovery in peripheral blood was accelerated in CD62L− T cell recipients comparing with that in the control T cell-depleted bone marrow recipients. Finally, we sought to determine whether these findings have any functional relevance in vivo. We evaluated the functional immune recovery by measuring the ability to inhibit the growth of a recipient-type leukemia/lymphoma cell line called BCL1 cells injected intravenously into the transplantation recipients on day +7. While six out of eight T cell-depleted bone marrow control mice developed tumor and died within 30 days after bone marrow transplantation, none of the CD62L− T cell recipients (n=12) developed tumor and all survived more than 60 days after transplantation. As expected, none of the CD62L− T cell recipients developed graft-versus-host disease. Our data indicate that CD62L− T cells promote new T cell generation by enhancing engraftment through overcoming host resistance. This promoting effect requires alloantigen recognition and is not mediated by “veto effect”. Our current focus is trying to determine how allogeneic CD62L− T cells overcome host resistance without causing graft-versus-host disease.

Delta-Like Notch Ligands Expressed By Host Non-Hematopoietic Radioresistant Cells Regulate Graft-Versus-Host Disease and Extrathymic T Cell Development After Bone Marrow Transplantation

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2003-2003
Author(s):  
Christen L Ebens ◽  
Jooho Chung ◽  
Ute Koch ◽  
Ivy T Tran ◽  
Ashley R Sandy-Sloat ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Marrow Transplantation ◽  
T Cell Development ◽  
Cell Development ◽  
Graft Versus Host ◽  
Cellular Source ◽  
Notch Ligands

Abstract Allogeneic bone marrow transplantation (allo-BMT) is limited by graft-versus-host disease (GVHD). We previously reported an essential role for Notch signaling in alloreactive T cells mediating GVHD after allo-BMT, demonstrating a profound decrease in GVHD incidence and severity with genetic Notch inhibition in donor T cells or systemic antibody-mediated blockade of Delta-like1 (Dll1) and Delta-like4 (Dll4) Notch ligands (Zhang et al., Blood 117(1), 2011; Sandy et al., J Immunol 190(11), 2013; Tran et al., JCI123(4), 2013). However, the cellular source of these critical Notch ligands remains unknown. While host hematopoietic antigen-presenting cells (APCs) seem a likely source, recent evidence indicates that these cells are not solely responsible for donor T cell activation in GVHD. We considered three alternative sources of Notch ligands: donor-derived hematopoietic cells, host hematopoietic APCs surviving lethal irradiation, and host non-hematopoietic radioresistant cells. To test these possibilities, we used complementary genetic and biochemical approaches to inactivate Dll1/Dll4 in specific compartments, or to provide ubiquitous systemic blockade of these ligands. Bone marrow (BM) chimeras were created by transplanting BM from poly(I:C)-induced Mx-Cre+ x Dll1f/fDll4f/f B6-CD45.2 mice into lethally irradiated B6-CD45.1 mice, generating chimeric mice without Dll1 and Dll4 only in the host hematopoietic compartment. After 14 weeks of reconstitution, CD45.2 → CD45.1 turnover was near complete in hematopoietic progenitors and professional APCs. Dll1/Dll4 excision was >99% by qPCR. WT (wild-type) BM chimeras subjected to MHC-mismatched allo-BMT (BALB/c → [Mx-Cre- x Dll1f/fDll4f/f (B6-CD45.2) → B6-SJL CD45.1]) had evidence of severe aGVHD and poor survival, as expected. Interestingly, BM chimera mice lacking Dll1 and Dll4 expression in the host hematopoietic compartment had equally severe GVHD and impaired survival following allo-BMT (BALB/c → [Mx-Cre+ x Dll1f/fDll4f/f (B6-CD45.2) → B6-SJL CD45.1]). In contrast, both WT and Dll1/Dll4-deficient BM chimeras treated with anti-Dll1/Dll4 antibodies (i.p. x4 over days 0-10 of transplant) had markedly decreased GVHD and improved survival (log rank Χ2=30.6, p<0.0001). Anti-Dll1/Dll4 antibodies blocked cytokine production by alloreactive T cells even after transplantation of purified T cells into irradiated recipients, ruling out dominant presentation of Notch ligands by donor-derived professional APCs. We are now backcrossing our Mx-Cre+ x Dll1f/fDll4f/f mice onto a BALB/c background to allow for simultaneous elimination of both donor and host hematopoietic Dll1 and Dll4 expression during allo-BMT. In addition to Notch-dependent regulation of GVHD, we studied immature pre-T cells that arise at extrathymic sites after BMT. We and others have observed Notch-dependent T cell development in lymphoid organs during early T cell reconstitution after transplantation (Lancrin et al., J Exp Med 195(7), 2002; Maillard et al., Blood 107(9), 2006; Holland et al., JCI122(12), 2013). Thus, this phenomenon is an alternative readout for exposure to Notch ligands in the post-BMT environment. Systemic blockade of Dll1 but not Dll4 with neutralizing antibodies completely blocked the development of these cells, indicating strict Dll1-dependence. In contrast, Dll1 elimination in host, donor or both host/donor hematopoietic compartments did not abrogate extrathymic pre-T cell development, consistent with a source of Notch ligands in host radioresistant cells. Altogether, these findings suggest that Notch ligands expressed by radio-resistant non-hematopoietic host tissues have important immunobiological functions during GVHD and T cell reconstitution. Identifying the cellular source of Delta-like ligands is critical to understand the effects of Notch signaling after bone marrow transplantation. Disclosures: Yan: Genentech, Inc: Employment. Siebel:Genentech, Inc: Employment.

Donor T cell–derived TNF is required for graft-versus-host disease and graft-versus-tumor activity after bone marrow transplantation

Blood ◽  
2003 ◽  
Vol 101 (6) ◽  
pp. 2440-2445 ◽  
Author(s):  
Cornelius Schmaltz ◽  
Onder Alpdogan ◽  
Stephanie J. Muriglan ◽  
Barry J. Kappel ◽  
Jimmy A. Rotolo ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Graft Versus Host Disease ◽  
Marrow Transplantation ◽  
F1 Hybrid ◽  
Host Disease ◽  
Graft Versus Host

Previous studies in murine bone marrow transplantation (BMT) models using neutralizing anti-tumor necrosis factor (TNF) antibodies or TNF receptor (TNFR)–deficient recipients have demonstrated that TNF can be involved in both graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL). TNF in these GVHD and GVL models was thought to be primarily produced by activated monocytes and macrophages, and the role of T cell–derived TNF was not determined. We used TNF−/− mice to study the specific role of TNF produced by donor T cells in a well-established parent-into-F1 hybrid model (C57BL/6J→C3FeB6F1/J). Recipients of TNF−/− T cells developed significantly less morbidity and mortality from GVHD than recipients of wild-type (wt) T cells. Histology of GVHD target organs revealed significantly less damage in thymus, small bowel, and large bowel, but not in liver or skin tissues from recipients of TNF−/− T cells. Recipients of TNF−/−T cells which were also inoculated with leukemia cells at the time of BMT showed increased mortality from leukemia when compared with recipients of wt cells. We found that TNF−/− T cells do not have intrinsic defects in vitro or in vivo in proliferation, IFN-γ production, or alloactivation. We could not detect TNF in the serum of our transplant recipients, suggesting that T cells contribute to GVHD and GVL via membrane-bound or locally released TNF.

scholarly journals Human HLA-specific T-cell clones with stable expression of a suicide gene: a possible tool to drive and control a graft-versus-host- graft- versus-leukemia reaction?

Blood ◽  
1996 ◽  
Vol 88 (3) ◽  
pp. 1098-1103 ◽  
Author(s):  
G Gallot ◽  
MM Hallet ◽  
J Gaschet ◽  
JF Moreau ◽  
R Vivien ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Bone Marrow Transplantation ◽  
T Cell ◽  
Marrow Transplantation ◽  
Host Recognition ◽  
T Cell Clones ◽  
Graft Versus Host ◽  
Graft Versus Leukemia ◽  
Cell Clones

Abstract Allogeneic bone marrow transplantation is still limited by the morbidity and mortality caused by graft-versus-host disease (GVHD), resulting from host recognition by donor T lymphocytes. It is possible to drastically reduce the T-cell content of the graft. However, transplanted T cells can also have a beneficial effect by graft enhancement and the graft-versus-leukemia effect. How can we keep the beneficial GVL effect while protecting the patient from possible GVHD? A recent report proposed the ex vivo transfer of the herpes simplex thymidine kinase (HSv-tk) gene into donor T cells before their infusion with hematopoietic stem cells. This procedure is expected to allow selective donor T-cell depletion with ganciclovir should GVHD occur, but it has two major drawbacks: reinjection of a fraction of untransfected T cells cannot be avoided and heterogeneity of the transfected population results in increased risks such as HSv-tk gene instability or dysfunction of some of the transfected T cell. Alternative approaches must be considered. We demonstrate here the feasibility of generating HSv-tk transfected HLA-specific CD4+ cytotoxic T-cell clonal populations, in which 100% of the cells have the HSv-tk gene inserted at a single site within their genome. These clones retained their specificity, their function, and their sensitivity to ganciclovir treatment. Our approach is not limited to bone marrow transplantation. Indeed, this procedure represents a useful alternative to retroviral gene transduction and is applicable to every circumstance where clinical use of gene modified T-cell clones is to be considered.

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