Functionally Mature CD4 and CD8 TCRαβ Cells Are Generated in OP9-DL1 Cultures from Human CD34+ Hematopoietic Cells

Coppernolle, Stefanie Van; Verstichel, Greet; Timmermans, Frank; Velghe, Imke; Vermijlen, David; Smedt, Magda De; Leclercq, Georges; Plum, Jean; Taghon, Tom; Vandekerckhove, Bart; Kerre, Tessa

doi:10.4049/jimmunol.0900714

Cited by 49 publications

(58 citation statements)

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“…Although two groups have reported the generation of mature single-positive T cells in OP9/DLL co-cultures [3,8], others failed [7]. Furthermore, questions remain about the functionality of these in vitro-matured T cells regarding their MHC-restriction and some phenotypic abnormalities [8]. In accordance with our data, Meek et al recently reported the same maturation arrest at the T/NK-progenitor cell level using a DLL-4 over-expressing stroma cell line [7].…”

Section: Cd7mentioning

confidence: 99%

Pre‐differentiated human committed T‐lymphoid progenitors promote peripheral T‐cell re‐constitution after stem cell transplantation in immunodeficient mice

et al. 2011

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T‐cell re‐constitution after allogeneic stem cell transplantation (alloSCT) is often dampened by the slow differentiation of human peripheral blood CD34+ (huCD34+) hematopoietic stem cells (HSCs) into mature T cells. This process may be accelerated by the co‐transfer of in vitro‐pre‐differentiated committed T/NK‐lymphoid progenitors (CTLPs). Here, we analysed the developmental potential of huCD34+ HSCs compared with CTLPs from a third‐party donor in a murine NOD‐scid IL2Rγnull model of humanised chimeric haematopoiesis. CTLPs (CD34+lin−CD45RA+CD7+) could be generated in vitro within 10 days upon co‐culture of huCD34+ or cord blood CD34+ (CB‐CD34) HSCs on murine OP9/N‐DLL‐1 stroma cells but not in a novel 3‐D cell‐culture matrix with DLL‐1low human stroma cells. In both in vitro systems, huCD34+ and CB‐CD34+ HSCs did not give rise to mature T cells. Upon transfer into 6‐wk‐old immune‐deficient mice, CTLPs alone did not engraft. However, transplantation of CTLPs together with huCD34+ HSCs resulted in rapid T‐cell engraftment in spleen, bone marrow and thymus at day 28. Strikingly, at this early time point mature T cells originated exclusively from CTLPs, whereas descendants of huCD34+ HSCs still expressed a T‐cell‐precursor phenotype (CD7+CD5+CD1a+/−). This strategy to enhance early T‐cell re‐constitution with ex vivo‐pre‐differentiated T‐lymphoid progenitors could bridge the gap until full T‐cell recovery in severely immunocompromised patients after allogeneic stem cell transplantation.

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Section: Cd7mentioning

confidence: 99%

Pre‐differentiated human committed T‐lymphoid progenitors promote peripheral T‐cell re‐constitution after stem cell transplantation in immunodeficient mice

et al. 2011

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“…RNA extraction, cDNA synthesis and RT-PCR were performed as described preciously. 18 The Figure 1 Phenotype of TCRgd þ thymocytes. Fresh postnatal thymocytes were labeled and analyzed by 7-color flow cytometry.…”

Section: Rna Isolation and Real-time (Rt)-pcrmentioning

confidence: 99%

“…18 An overview of the antibodies that were used can be found in Supplementary Materials. Flow cytometric analysis was performed using a LSR II Cytometer (BD Biosciences).…”

Section: Flow Cytometrymentioning

confidence: 99%

“…Cells harvested from OP9-DL1 co-cultures were either directly transferred to a feeder cell mixture 18 or were first sorted for subpopulations.…”

Section: T-cell Expansionmentioning

confidence: 99%

“…Co-cultures were performed as previously described. 18 In some experiments, different concentrations of the g-secretase inhibitor 7 N-[N-(3,5-difluorophenyl-L-alanyl)]-S-phenyl-glycine t-butyl ester (DAPT) (Peptides International, Louisville, KY, USA), diluted in dimethyl sulfoxide (Serva Electrophoresis, Heidelberg, Germany) were added. Sorted gd cell fractions were either immediately seeded on OP9-DL1 or OP9-GFP, or first labeled with PKH (named after the discoverer, Paul Karl Horan) (Sigma-Aldrich, Saint Louis, MI, USA), according to the manufacturers' instructions.…”

Section: Op9-dl1 Co-culturesmentioning

confidence: 99%

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Notch induces human T-cell receptor γδ+ thymocytes to differentiate along a parallel, highly proliferative and bipotent CD4 CD8 double-positive pathway

et al. 2011

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In wild-type mice, T-cell receptor (TCR) cd þ cells differentiate along a CD4 CD8 double-negative (DN) pathway whereas TCRab þ cells differentiate along the double-positive (DP) pathway. In the human postnatal thymus (PNT), DN, DP and single-positive (SP) TCRcd þ populations are present. Here, the precursor -progeny relationship of the various PNT TCRcd þ populations was studied and the role of the DP TCRcd þ population during T-cell differentiation was elucidated. We demonstrate that human TCRcd þ cells differentiate along two pathways downstream from an immature CD1 þ DN TCRcd þ precursor: a Notch-independent DN pathway generating mature DN and CD8aa SP TCRcd þ cells, and a Notch-dependent, highly proliferative DP pathway generating immature CD4 SP and subsequently DP TCRcd þ populations. DP TCRcd þ cells are actively rearranging the TCRa locus, and differentiate to TCR À DP cells, to CD8ab SP TCRcd þ cells and to TCRab þ cells. Finally, we show that the cd subset of T-cell acute lymphoblastic leukemias (T-ALL) consists mainly of CD4 SP or DP phenotypes carrying significantly more activating Notch mutations than DN T-ALL. The latter suggests that activating Notch mutations in TCRcd þ thymocytes induce proliferation and differentiation along the DP pathway in vivo.

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In Vitro T-Cell Generation From Adult, Embryonic, and Induced Pluripotent Stem Cells: Many Roads to One Destination

et al. 2015

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T lymphocytes are critical mediators of the adaptive immune system and have the capacity to serve as therapeutic agents in the areas of transplant and cancer immunotherapy. While T cells can be isolated and expanded from patients, T cells derived in vitro from both hematopoietic stem/progenitor cells (HSPCs) and human pluripotent stem cells (hPSCs) offer great potential advantages in generating a self-renewing source of T cells that can be readily genetically modified. T-cell differentiation in vivo is a complex process requiring tightly regulated signals; providing the correct signals in vitro to induce T-cell lineage commitment followed by their development into mature, functional, single positive T cells, is similarly complex. In this review, we discuss current methods for the in vitro derivation of T cells from murine and human HSPCs and hPSCs that use feeder-cell and feeder-cell-free systems. Furthermore, we explore their potential for adoption for use in T-cell-based therapies.

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Functionally Mature CD4 and CD8 TCRαβ Cells Are Generated in OP9-DL1 Cultures from Human CD34+ Hematopoietic Cells

Abstract: Material

Cited by 49 publications

References 50 publications

Pre‐differentiated human committed T‐lymphoid progenitors promote peripheral T‐cell re‐constitution after stem cell transplantation in immunodeficient mice

Pre‐differentiated human committed T‐lymphoid progenitors promote peripheral T‐cell re‐constitution after stem cell transplantation in immunodeficient mice

Notch induces human T-cell receptor γδ+ thymocytes to differentiate along a parallel, highly proliferative and bipotent CD4 CD8 double-positive pathway

In Vitro T-Cell Generation From Adult, Embryonic, and Induced Pluripotent Stem Cells: Many Roads to One Destination

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