The impact of aging on memory T cell phenotype and function in the human bone marrow

In order to study extrathymic differentiationin vitro, CD7+CD3-lymphocytes were sorted from normal human bone marrow and cultured under conditions of limiting dilution together with irradiated pooled allogeneic peripheral blood mononuclear cells (PBMC) and phytohemagglutinin (PHA) in the presence of 1000 U/ml of interleukin-2 (IL-2). One clone was obtained that failed to react with monoclonal antibody (mAb) TCRδ1 (TCRγ/δ-specific) or WT31 (TCR2,α/β-specific). From day 35 through day 74 in culture, the surface phenotype of this clone evolved into CD3+, CD4+, CD8-, TCR2+, TCR1-, and was further characterized as CD2+, CD45RO+, CD16-, and CD56-. The presence of mRNA for TCRαandγbut not ,andγchains was confirmed by Northern blotting. Accessory cell-dependent autocrine proliferative responses to PHA (most likely driven by IL-2) were initially absent, but became measurable at the same time as the TCR was acquired. However, in the absence of PHA, the clone failed to respond to a panel of homozygous B-cell lines representing the majority of MHC class II alleles. Autoreactivity was also not demonstrable. Cytotoxicity was limited to MHC unrestricted “natural killer (NK)-like” lysis of K562 target cells, with no autocytotoxicity detected. Tle NK-like lysis diminished over time in parallel with the acquisition of surface TCR. The cloned cells were not suppressive for mature lymphocyte proliferation. After stimulation, the cells secreted tumor necrosis factorαand granulocyte/macrophage colony-stimulating factor (GM-CSF) detected by immunoassays, and T-cell growth factors, most likely IL-2, as detected by bioassays. Polymerase chain-reaction methods demonstrated the presence of mRNA for IL-2, IL-3, IL-4, IL-9, interferon-δ, and GM-CSF in these cells after stimulation with PHA and B-LCL.These results suggest that cells with the phenotype and some functional characteristics of mature T lymphocytes can evolve extrathymicallyin vitrofrom T-cell precursors sorted from normal human bone marrow.

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Human bone marrow adipocytes support dexamethasone-induced osteoclast differentiation and function through RANKL expression

Biomedical Research ◽

10.2220/biomedres.32.37 ◽

2011 ◽

Vol 32 (1) ◽

pp. 37-44 ◽

Cited By ~ 31

Author(s):

Hisataka Goto ◽

Makoto Osaki ◽

Tatsuya Fukushima ◽

Kazutaka Sakamoto ◽

Akira Hozumi ◽

...

Keyword(s):

Bone Marrow ◽

Osteoclast Differentiation ◽

Human Bone ◽

Human Bone Marrow ◽

Rankl Expression ◽

Bone Marrow Adipocytes ◽

And Function

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Manufacturing Differences Affect Human Bone Marrow Stromal Cell Characteristics and Function: Comparison of Production Methods and Products from Multiple Centers

Scientific Reports ◽

10.1038/srep46731 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 38

Author(s):

Shutong Liu ◽

Luis F. de Castro ◽

Ping Jin ◽

Sara Civini ◽

Jiaqiang Ren ◽

...

Keyword(s):

Bone Marrow ◽

Stromal Cell ◽

Bone Marrow Stromal Cell ◽

Human Bone ◽

Human Bone Marrow ◽

Production Methods ◽

Marrow Stromal Cell ◽

And Function

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A Human Lymphoid Progenitor Capable of Circulating and Seeding the Thymus.

Blood ◽

10.1182/blood.v110.11.2216.2216 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2216-2216

Author(s):

Emmanuelle M. Six ◽

Delphine Bonhomme ◽

Marta Monteiro ◽

Kheira Beldjord ◽

Alexandrine Garrigue ◽

...

Keyword(s):

Gene Expression ◽

Bone Marrow ◽

T Cell ◽

T Cell Development ◽

Human Bone ◽

Human Bone Marrow ◽

Rt Pcr ◽

Pcr Assay ◽

First Time

Abstract Identification of a thymus-seeding progenitor originating from human bone marrow constitutes a key milestone in understanding and correcting defects in T-cell development. Here, we report the characterization of a novel human bone marrow lymphoid-restricted subpopulation which is part of the lineage-negative CD34+CD10+ progenitor population and which can be distinguished from B-cell-committed precursors by the absence of CD24 expression. We demonstrated that these Lin-CD34+CD10+CD24- progenitors lack myeloid and erythroid potential but can generate B, T and NK lymphocytes following culture on MS5 or OP9-hDelta1 stroma. The gene expression profile of this population, analyzed by a multiplex RT-PCR assay, revealed co-expression of RAG1, TdT, PAX5, CD3ε and IL-7Rα. These progenitors are not only present in the bone marrow but also in the blood throughout life, suggesting an ability to circulate. Moreover we showed that the Lin-CD34+CD10+CD24- cells also correspond to the most immature population of the thymus which gives rise to Lin-CD34+CD7+ T-cell precursors. Taken as a whole these findings unravel for the first time the existence of a postnatal lymphoid-restricted population which is capable of migrating from the bone marrow to the thymus.

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A Common Progenitor for Macrophages, Osteoclasts and Dendritic Cells Newly Discovered in Human Bone Marrow and Cord Blood Yields Dendritic Cells with T-Cell Priming Ability

Blood ◽

10.1182/blood.v126.23.645.645 ◽

2015 ◽

Vol 126 (23) ◽

pp. 645-645

Author(s):

Yanling Xiao ◽

Joanna Aleksandra Grabowska ◽

Riccardo Mezzadra ◽

Maarten J. van Tol ◽

Arjan C Lankester ◽

...

Keyword(s):

Bone Marrow ◽

Cell Proliferation ◽

Dendritic Cells ◽

Stem Cell ◽

T Cell ◽

Cord Blood ◽

Human Bone ◽

Human Bone Marrow ◽

T Cell Proliferation ◽

T Cell Priming

Abstract Dendritic cells (DC) have potent antigen-presentation and T-cell priming ability and therefore hold great promise in cancer immunotherapy. However, DC vaccination has not yet delivered a reliable clinical response rate, despite great efforts. Since primary DCs are rare (up 0.2-1.5% of circulating leukocytes), therapeutic DCs are generally derived from peripheral blood monocytes by culture with GM-CSF (moDC). Vaccines composed of moDC loaded with tumor antigens can induce potent and long-lasting tumour-specific immune responses in patients, but such positive results are infrequent and unpredictable. To improve success rate, research has focused on moDC culture regimens, antigen loading and activation strategies and methods of DC injection. Nevertheless, to date clinical trials using moDC have not yielded statistically significant treatment benefits over conventional strategies. Current attention has therefore shifted to the rare primary DCs that circulate in the blood under homeostatic conditions. Knowing the identity of the precursors of these DCs may facilitate the ex-vivo or in-vivo generation of DCs via the homeostatic pathway, potentially yielding DCs with optimal T cell priming ability. We (Xiao et al. Stem Cell Rep. 2015) and others (Lee et al. J. Exp. Med. 2015) have recently identified a population with DC progenitor potential in human bone marrow and cord blood, respectively. This population can be isolated on basis of a CD34+ c-KIT+ FLT3+ IL3Rαhigh phenotype and is furthermore Lin- CD10- CD11b- CD45RA+ CD38+. We have shown that this population is highly enriched for or identical to a common progenitor (P) of macrophages (M), osteoclasts (O) and DCs (D) and termed it MODP. We also identified the progenitor directly upstream from the MODP that still has granulocyte (G) differentiation potential and termed it GMODP. We hypothesized that DCs generated from GMODP or MODP under homeostatic conditions would have superb T-cell priming capacity. To examine this, the progenitors were sorted by flow cytometry from human bone marrow or cord blood and cultured with Flt3 ligand, M-CSF and IL-3 to generate DCs. We also tested the effect of a mensenchymal stem cell (MSC) feeder layer. Within 2-3 weeks of culture, 1000 DC progenitors generated approximately 150,000-250,000 DCs. Co-culture with MSC increased DC output significantly, at least 2 fold. The progenitor-derived DCs could be discerned into CD141+ conventional (c)DC, CD1c+ cDC and CD303+ plasmacytoid (p)DC. To study T-cell priming capacity of progenitor-derived DCs, we set up an in vitro DC-T co-culture assay. CD141+ cDC, CD1c+ cDC and CD303+ pDC were generated from GMODP or MODP of HLA-A2+ donors, flow cytometrically purified, activated with lipopolysaccharide and loaded with MART-126-35 peptide that represents a melanoma-derived tumor antigen. Primary T cells from peripheral blood of unrelated donors were retrovirally transduced to express a T cell antigen receptor (TCR) ab specific for the HLA-A2/MART-126-35 peptide complex. The ability of the DCs to prime a T-cell response was read out by antigen-specific CD8+ T cell proliferation. All DC subsets were able to induce MART-1 specific T cell proliferation, with the CD1c+ cDCs being most potent and the CD303+ pDC being least potent. In conclusion: We have established a culture method to derive DCs with T-cell priming ability from a newly identified DC progenitor. These results are of value for improvement of DC-based immunotherapy. Disclosures No relevant conflicts of interest to declare.

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