Identification of an NK/T cell–restricted progenitor in adult bone marrow contributing to bone marrow– and thymic-dependent NK cells

Charoudeh, Hojjatollah Nozad; Tang, Yanjuan; Cheng, Min; Cilio, Corrado; Jacobsen, Sten Eirik W.; Sitnicka, Ewa

doi:10.1182/blood-2009-10-247130

Cited by 40 publications

(55 citation statements)

References 47 publications

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“…2 This population is likely to be heterogeneous because only 8%-40% of NKPs are reported to have unilineage NK-cell potential. 2,3,14 To test whether ID2 expression can be used to identify committed NK-cell progenitors, we analyzed the NKP population by using mice that have GFP introduced into the Id2 locus (ID2 gfp ). Flow cytometric analysis showed that lin Ϫ CD122 ϩ NK1.1 Ϫ CD49b Ϫ NKPs could be subdivided into low and high ID2-expressing cells ( Figure 1A; supplemental Figure 1A).…”

Section: Resultsmentioning

confidence: 99%

“…Strikingly, only ID2 gfp -high NKPs efficiently generated NK1.1 ϩ CD49b ϩ NK cells (Table 1). 13 The authors of a recent report 14 showed that the NKP population consists of NK cellrestricted and NK/T bipotent precursor cells. Although we obtained similar results with unfractionated NKPs, we found that only NK-cell potential was enriched in ID2 gfp -high cells (data not shown).…”

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confidence: 99%

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Identification of the earliest NK-cell precursor in the mouse BM

Carotta

Pang

Nutt

et al. 2011

Blood

155

167

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IntroductionAlthough the developmental stages leading to committed B and T cells have been dissected in great detail, 1 the developmental intermediates downstream of the common lymphoid progenitor (CLP) leading to a committed natural killer (NK)-cell precursor remain poorly defined. The earliest committed NK-cell progenitor (NKP) identified to date is characterized through the expression of the IL-2 receptor ␤ chain (CD122) and the lack of pan-NK-cell surface markers (NK1.1 and CD49b), but this population is likely to be heterogeneous because only a minority of cells give rise to NK cells in in vitro assays. 2,3 The transcription factor inhibitor of DNA binding 2 (ID2) is essential for NK-cell development. In its absence, fetal and adult NK-cell development is blocked at an early stage. [4][5][6][7] Id2 inhibits E proteins in NK cells and is itself regulated by E4BP4, which is also required for the differentiation of NKPs to immature NK cells. 8,9 Using a reporter line in which the green fluorescent protein (GFP) gene has been introduced into the Id2 locus, we report the identification of the earliest committed NK-cell precursor in adult BM. By analogy to B-cell development, we termed these pre-pro NK cells, and they express high levels of ID2, IL-7R␣ (CD127), and lack expression of most, but not all, NK-cell markers. Furthermore, we demonstrate that ID2 gfp expression can be used to isolate the committed NK-cell progenitors within the heterogeneous NKP population. Methods MiceWe generated ID2 gfp reporter mice by inserting an internal ribosome entry site-GFP cassette into the 3Ј untranslated region of the Id2 locus (G.T.B., S.C., and S.L.N. manuscript submitted April 2011). PU.1 gfp and Rag1 gfp mice have been described previously. 10,11 All mice were maintained under specific pathogen-free conditions at the Walter and Eliza Hall Institute according to institute guidelines. Antibodies, cell analysis, and sortingLineage (lin) marker-positive BM cells were depleted and analyzed by flow cytometry as described in supplemental Methods (available on the Blood Web site; see the Supplemental Materials link at the top of the online article). Cell cultureTo determine precursor frequencies, sorted progenitors cells were seeded in limiting dilution on OP9 or OP9-DL1 cells in the presence of 2% IL-7 supernatant and 5 ng/mL FMS-like tyrosine kinase 3 (Flt3) ligand for Band T-cell development, respectively, and on OP9 cells plus 0.5% IL-7 supernatant and 50 ng/mL human IL-15 (R&D Systems) for NK-cell differentiation as described. 12 After 7-10 days, wells were scored for the presence of colonies. Colonies were harvested for flow cytometric analysis, and precursor frequency was calculated according to Poisson statistics with the use of ELDA (extreme limiting dilution analysis) software Version 1.0 (http://bioinf.wehi.edu.au/software/elda/). 13 Results and discussionThe earliest committed NKP identified to date expresses CD122 but not the pan-NK cell-specific markers NK1.1 and CD49b. 2 This population is likely to be heterogene...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Identification of the earliest NK-cell precursor in the mouse BM

Carotta

Pang

Nutt

et al. 2011

Blood

155

167

View full text Add to dashboard Cite

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“…During lymphopoiesis, both fetal and adult CLPs can lose the potential to give rise to T cells before they lose NK potential. Although in the most common hierarchical models of hematopoiesis NK cells appear more closely related to the T cell lineage (49)(50)(51)(52)(53)(54) embryos generated T cells, suggesting that these two proteins compensated for each other in their function. Accordingly, and similarly to what is seen in NK precursors (47), Id3 mRNA expression was higher in CLPs from Id2 2/2 mice (data not shown).…”

Section: Discussionmentioning

confidence: 99%

Inhibitors of DNA Binding Proteins Restrict T Cell Potential by Repressing Notch1 Expression in Flt3-Negative Common Lymphoid Progenitors

Sousa

Berthault

Granato

et al. 2012

The Journal of Immunology

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Lineage commitment is regulated during hematopoiesis, with stepwise loss of differentiation potential ultimately resulting in lineage commitment. In this study we describe a novel population of B/NK bipotent precursors among common lymphoid progenitors in the fetal liver and the bone marrow. The absence of T cell precursor potential, both in vivo and in vitro, is due to low Notch1 expression and secondary to inhibition of E2A activity by members of the inhibitor of DNA binding (Id) protein family. Our results demonstrate a new, Id protein-dependent, molecular mechanism of Notch1 repression, operative in both fetal and adult common lymphoid progenitors, where T cell potential is selectively inhibited without affecting either the B or NK programs. This study identifies Id proteins as negative regulators of T cell specification, before B and NK commitment, and provides important insights into the transcriptional networks orchestrating hematopoiesis.

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“…Although thymic NK cells can develop in the absence of signals essential for T cell development, it remains possible that thymic NK cells may derive from thymic seeding of the recently described early bipotent NK/T progenitor present in the BM (6). In contrast, DN2 thymocytes, although exhibiting NK cell potential in different experimental systems (27), appear to represent only a marginal substrate for the development of thymic NK cells, at least under mphysiologic conditions, as the latter can develop in absence of all T cell precursors and show little evidence of Ag-receptor rearrangements.…”

Section: The Journal Of Immunology 4995mentioning

confidence: 99%

“…The Takei laboratory reported TCRg rearrangements in a large fraction of NK cells in the thymus and lymph nodes (4,5) and suggested that thymic NK cells might share a precursor stage with T cells and thus represent failed T cell precursors (4). In addition, recent data showed that the population of NK cell progenitor cells in the BM encompasses cells with not only NK potential but also T as well as NK/T bipotent precursor cells (6). The relationship of thymic NK cells to classical NK cells, innate T lymphocytes (gd T cells and NKT cells), and mainstream ab T cells remains unclear.…”

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confidence: 99%

Cutting Edge: Thymic NK Cells Develop Independently from T Cell Precursors

Ribeiro

Hasan

Wilson

et al. 2010

The Journal of Immunology

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Although NK cells in the mouse are thought to develop in the bone marrow, a small population of NK cells in the thymus has been shown to derive from a GATA3-dependent pathway. Characteristically, thymic NK cells express CD127 and few Ly49 molecules and lack CD11b. Because these NK cells develop in the thymus, the question of their relationship to the T cell lineage has been raised. Using several different mouse models, we find that unlike T cells, thymic NK cells are not the progeny of Rorc-expressing progenitors and do not express Rag2 or rearrange the TCRγ locus. We further demonstrate that thymic NK cells develop independently of the Notch signaling pathway, supporting the idea that thymic NK cells represent bona fide NK cells that can develop independently of all T cell precursors.

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Identification of an NK/T cell–restricted progenitor in adult bone marrow contributing to bone marrow– and thymic-dependent NK cells

Cited by 40 publications

References 47 publications

Identification of the earliest NK-cell precursor in the mouse BM

Identification of the earliest NK-cell precursor in the mouse BM

Inhibitors of DNA Binding Proteins Restrict T Cell Potential by Repressing Notch1 Expression in Flt3-Negative Common Lymphoid Progenitors

Cutting Edge: Thymic NK Cells Develop Independently from T Cell Precursors

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