Human CD8 T cells generated in vitro from hematopoietic stem cells are functionally mature

Awong, Génève; Herer, Elaine; Motte-Mohs, Ross N. La; Zúñiga‐Pflücker, Juan Carlos

doi:10.1186/1471-2172-12-22

Cited by 43 publications

(45 citation statements)

References 33 publications

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“…Cultures were analyzed for the presence of CD1a+CD7+ cells, previously identified as early T cells, and CD4+CD8+ DP T cells (Supplementary Figure 1B, 1C, and Figure 2B, 2C). 10,12 At day 16, we found about twice as many CD1a+CD7+ cells in the Notch ligand conditions (2.5 µg/mL Fc-DLL1) compared to the no ligand control (Supplementary Figure 1B). After 25 days of culture with Fc-DLL1, the cultures had about 10 times as many CD7+CD1a cells compared to the control (Figure 2B).…”

Section: Resultsmentioning

confidence: 87%

“…Specifically, to induce T lymphoid lineage commitment, ES cells or HSCs can be co-cultured with OP9-DL1 cells, bone marrow-derived stromal cells retrovirally transfected to stably express the Notch ligand DLL1. 10–13 The OP9-DL1 co-culture system is now an established method for differentiating mouse and human hematopoietic stem cells into early T cells as well as CD8+ SP T cells; however, the generation of human antigen-specific, functional T cells without retroviral transfection of specific TCRs has not been reported. 10–13,21,22,33 Furthermore, large scale production of therapeutic T cells for eventual clinical application via co-culture with retrovirally transfected cells is difficult, both from pharmaceutical and regulatory perspectives.…”

Section: Discussionmentioning

confidence: 99%

“…Retrovirally-transfected, mouse bone marrow-derived stromal cells (OP9) that stably express the Notch ligands, DLL1 (OP9-DL1) or DLL4 (OP9-DL4), are capable of supporting the differentiation of mouse hematopoietic, embryonic, and induced pluripotent stem cells, as well as human hematopoietic stem cells into early T cells and CD8+ SP T cells. 10–13 Recent studies have also shown that plate-bound Notch ligands and a defined combination of soluble cytokines induce early T cell development from mouse Lin-c-kit+Sca-1+ or human CD34+ HSCs. 14–17 Our group has previously shown that culturing mouse Lin-c-kit+Sca-1+ HSCs with DLL4-functionalized microbeads in an insert co-culture system using OP9-DL1 cells can induce early T lineage commitment and differentiation without direct stromal cell contact.…”

Section: Introductionmentioning

confidence: 99%

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Generation of Functional, Antigen-Specific CD8+ Human T Cells from Cord Blood Stem Cells Using Exogenous Notch and Tetramer-TCR Signaling

2014

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In vitro differentiation of mouse and human stem cells into early T cells has been successfully demonstrated using artificial Notch signaling systems. However, generation of mature, antigen-specific, functional T cells, directly from human stem cells has remained elusive, except when using stromal co-culture of stem cells retrovirally transfected with antigen-specific T cell receptors (TCRs). Here we show that human umbilical cord blood (UCB)-derived CD34+CD38−/low hematopoietic stem cells (HSCs) can be successfully differentiated into functional, antigen-specific cytotoxic CD8+ T cells without direct stromal co-culture or retroviral TCR transfection. Surface-immobilized Notch ligands (DLL1) and stromal cell conditioned medium successfully induced the development of CD1a+CD7+ and CD4+CD8+ early T cells. These cells, upon continued culture with cytomegalovirus (CMV) or Influenza-A virus epitope-loaded HLA-A*0201 tetramers, resulted in the generation of a polyclonal population of CMV-specific or Influenza-specific CD8+ T cells respectively. Upon further activation with antigen-loaded target cells, these antigen-specific, stem cell-derived T cells exhibited cytolytic functionality, specifically CD107a surface mobilization, IFNγ production, and Granzyme B secretion. Such scalable, in vitro generation of functional, antigen-specific human T cells from human stem cells could eventually provide a readily available cell source for adoptive transfer immunotherapies and also allow better understanding of human T cell development.

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Section: Resultsmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Generation of Functional, Antigen-Specific CD8+ Human T Cells from Cord Blood Stem Cells Using Exogenous Notch and Tetramer-TCR Signaling

2014

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“…Further differentiation on OP9-DL cells, yields mature SP cells, and similar to the mouse system with a strong bias towards the CD8 + SP fate. In vitro-generated CD8 + SPs, the majority of which are CD3 + TCRαβ + , show similar levels of CD8-specific transcription factors when compared to ex-vivo human UCB-derived T cells (Awong et al 2011). When activated through CD3/CD28 stimulation, similar to CD8 cytotoxic cells, the in vitro-derived CD8 cells acquire the expression of the cytotoxic agent Granzyme B and release the effector cytokine, Interferon-γ.…”

Section: Human T Cell Generation In Vitromentioning

confidence: 92%

Artificial Thymus: Recreating Microenvironmental Cues to Direct T Cell Differentiation and Thymic Regeneration

Mohtashami

Shukla

Zandstra

et al. 2016

Synthetic Immunology

Self Cite

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The thymus supports the development and differentiation of T cells, which are a central component of the mammalian adaptive immune system. From entry of hematopoietic progenitor cells into the thymus to their complex maturation sequence into naïve T cells, the role of the thymic microenvironment has been the subject of intense study. A pivotal aspect of this process is the activation of Notch receptors on progenitors by Delta-like (Dll) ligands present on thymic epithelial cells. Thus far, two approaches have been taken to create an artificial thymus, or mimic thymic function. One involves an in vitro cell-based system in which several key components are provided, including Dll and cytokines, to induce and support T cell lineage differentiation. The gold standard approach makes use of a bone marrow-derived cell line (OP9), ectopically expressing Dll (OP9-DL). A related method involves an in vitro cell-free system that provides a similar set of required signaling components. The second approach involves the generation of organized thymic tissue, which can be achieved by direct reprogramming of another cell type or via the differentiation of pluripotent stem cells (PSCs), either by the ectopic expression of the thymic master regulatory gene, FoxN1, in fibroblasts or by inducing the differentiation of PSCs using developmental cues, respectively. These approaches share a similar goal, to generate T cells from different sources of stem cells. However, the former takes advantage of cellular or molecular drivers of T-lineage differentiation, while the latter is focused on creating thymic tissues that would support T cell development.

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“…To this end, it might be advantageous to expand LAA-specific T cells from ex vivogenerated naive autologous T cells that have not been subjected to immunoediting in vivo. 112 However, T cells from a healthy allogeneic donor may have an advantage over autologous T cells: healthy subjects that have not been subjected to chemotherapy may have a superior thymic function and a more diversified T-cell repertoire. 113 …”

Section: Laa-targeted Atci Of Leukemiamentioning

confidence: 99%

Next-generation leukemia immunotherapy

Vincent

Roy

Perreault

2011

Blood

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Allogeneic hematopoietic cell transplantation led to the discovery of the allogeneic GVL effect, which remains the most convincing evidence that immune cells can cure cancer in humans. However, despite its great paradigmatic and clinical relevance, induction of GVL by conventional allogeneic hematopoietic cell transplantation remains a quite rudimentary form of leukemia immunotherapy. It is toxic and its efficacy is far from optimal. It is therefore sobering that since the discovery of the GVL effect 3 decades ago, the way GVL is induced and manipulated has practically not changed. Preclinical and clinical studies suggest that injection of T cells primed against a single Ag present on neoplastic cells could enhance the GVL effect without causing any GVHD. We therefore contend that Ag-targeted adoptive T-cell immunotherapy represents the future of leukemia immunotherapy, and we discuss the specific strategies that ought to be evaluated to reach this goal. Differences between these strategies hinge on 2 key elements: IntroductionThe GVL reaction refers to the ability of donor immune cells to eliminate host leukemic cells after allogeneic hematopoietic cell transplantation (AHCT). In 1956, Barnes et al 1 were the first to report cure of leukemia in mice after total body irradiation and AHCT. Further studies of GVL in animal models were pioneered by Bortin et al 2 in the 1970s. 3 The relevance of the GVL reaction in humans was established by the Seattle group in 1979, and key insights into its mechanisms were reported in a landmark study from the International Bone Marrow Transplant Registry in 1990. 4,5 Strikingly, the latter study that was based on data from 2254 subjects treated by 142 teams showed that GVL was abrogated if T cells were depleted from the graft or if the AHCT donor was an identical twin. 5 On the basis of these data, it was therefore inferred that GVL depended on donor T cells and on the existence of histocompatibility differences between the donor and its recipient (absent among identical twins). Because the International Bone Marrow Transplant Registry study involved HLA-identical siblings, the sole histocompatibility differences between donors and recipients were minor histocompatibility Ags (MiHAs). 6 The molecular nature of MiHAs was elucidated in 1990, and the first human MiHA was sequenced in 1995. 7,8 Increasing recognition that cure after AHCT for leukemia is largely because of the GVL effect led to the introduction of nonmyeloablative conditioning regimens and donor lymphocyte infusions (DLIs). 9,10 Remarkably, DLI can eradicate Յ 10 12 neoplastic cells in patients with chronic myelogenous leukemia, 11,12 and the allogeneic GVL effect represents the most convincing evidence that immunotherapy can cure human neoplasias. 13,14 Nowadays, AHCT can be viewed primarily as a quest for the GVL effect. 12 In AHC transplant recipients, the effect of GVL on leukemia-free survival is so important that it supersedes the deleterious effect of GVHD and protracted immunodeficiency. After conven...

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Human CD8 T cells generated in vitro from hematopoietic stem cells are functionally mature

Cited by 43 publications

References 33 publications

Generation of Functional, Antigen-Specific CD8+ Human T Cells from Cord Blood Stem Cells Using Exogenous Notch and Tetramer-TCR Signaling

Generation of Functional, Antigen-Specific CD8+ Human T Cells from Cord Blood Stem Cells Using Exogenous Notch and Tetramer-TCR Signaling

Artificial Thymus: Recreating Microenvironmental Cues to Direct T Cell Differentiation and Thymic Regeneration

Next-generation leukemia immunotherapy

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