scholarly journals Human Bone Marrow-Derived Mesenchymal Stromal Cells Differentially Inhibit Cytokine Production by Peripheral Blood Monocytes Subpopulations and Myeloid Dendritic Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-15 ◽  
Author(s):  
Paula Laranjeira ◽  
Joana Gomes ◽  
Susana Pedreiro ◽  
Monia Pedrosa ◽  
Antonio Martinho ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Dendritic Cells ◽  
Protein Expression ◽  
Peripheral Blood ◽  
Immune Cell ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Mrna Levels ◽  
Myeloid Dendritic Cells ◽  
Experimental Conditions

The immunosuppressive properties of mesenchymal stromal/stem cells (MSC) rendered them an attractive therapeutic approach for immune disorders and an increasing body of evidence demonstrated their clinical value. However, the influence of MSC on the function of specific immune cell populations, namely, monocyte subpopulations, is not well elucidated. Here, we investigated the influence of human bone marrow MSC on the cytokine and chemokine expression by peripheral blood classical, intermediate and nonclassical monocytes, and myeloid dendritic cells (mDC), stimulated with lipopolysaccharide plus interferon (IFN)γ. We found that MSC effectively inhibit tumor necrosis factor- (TNF-)αand macrophage inflammatory protein- (MIP-) 1βprotein expression in monocytes and mDC, without suppressing CCR7 and CD83 protein expression. Interestingly, mDC exhibited the highest degree of inhibition, for both TNF-αand MIP-1β, whereas the reduction of TNF-αexpression was less marked for nonclassical monocytes. Similarly, MSC decreased mRNA levels of interleukin- (IL-) 1βand IL-6 in classical monocytes, CCL3, CCL5, CXCL9, and CXCL10 in classical and nonclassical monocytes, and IL-1βand CXCL10 in mDC. MSC do not impair the expression of maturation markers in monocytes and mDC under our experimental conditions; nevertheless, they hamper the proinflammatory function of monocytes and mDC, which may impede the development of inflammatory immune responses.

scholarly journals Identification of hematopoietic progenitors of macrophages and dendritic Langerhans cells (DL-CFU) in human bone marrow and peripheral blood

Blood ◽  
1990 ◽  
Vol 76 (6) ◽  
pp. 1139-1149 ◽  
Author(s):  
CD Reid ◽  
PR Fryer ◽  
C Clifford ◽  
A Kirk ◽  
J Tikerpae ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Dendritic Cells ◽  
Peripheral Blood ◽  
Langerhans Cells ◽  
Mononuclear Cells ◽  
Cultured Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Nonspecific Esterase ◽  
Hla Dq

Abstract Colonies of cells with distinctive dendritic appearance were observed in methylcellulose cultures of human bone marrow and peripheral blood mononuclear cells (PBMC). Such cells appeared alone in colonies of less than 50 cells, together with macrophages in mixed colonies and also within clusters of T lymphocytes at high culture cell numbers. The morphologic resemblance to lymphoid dendritic cells was confirmed by electron microscopy and the cells were distinguished from macrophages by immunoenzymatic and immunogold labeling with monoclonal antibodies (MoAbs). Like macrophages they were HLA-DR+ and CD4+. However, they lacked nonspecific esterase and the macrophage cytoplasmic marker Y1/82A. Most strikingly, cells were strongly HLA-DQ+ and expressed CD1a (T6), which is characteristic of skin Langerhans cells. Their functional similarity to lymphoid dendritic cells was demonstrated by their ability to stimulate allogeneic mixed leukocyte reactions. Dendritic cell colony numbers were estimated in both bone marrow and peripheral blood of controls and in leukemia and lymphoma patients before and after chemotherapy. Colony numbers were low in control blood and in patients before treatment (less than 1.0 to 3.7/10(5) cells). However, during hematopoietic recovery the mean value increased to 37.5/10(5) cells and this increase correlated closely with the observed increase in circulating colony forming unit-granulocyte macrophage (CFU- GM) in individual patients. Autoradiographic studies demonstrated mitotic activity within CD1a+ colonies and a linear relationship between cultured cells and both pure and mixed colonies was consistent with their derivation from a single precursor. These data indicate that a novel hematopoietic progenitor of dendritic/Langerhans cells (DL-CFU) may now be identified in a clonal assay system and suggest a probable common progenitor for these cells and macrophages.

scholarly journals Identification of hematopoietic progenitors of macrophages and dendritic Langerhans cells (DL-CFU) in human bone marrow and peripheral blood

Blood ◽  
1990 ◽  
Vol 76 (6) ◽  
pp. 1139-1149 ◽  
Author(s):  
CD Reid ◽  
PR Fryer ◽  
C Clifford ◽  
A Kirk ◽  
J Tikerpae ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Dendritic Cells ◽  
Peripheral Blood ◽  
Langerhans Cells ◽  
Mononuclear Cells ◽  
Cultured Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Nonspecific Esterase ◽  
Hla Dq

Colonies of cells with distinctive dendritic appearance were observed in methylcellulose cultures of human bone marrow and peripheral blood mononuclear cells (PBMC). Such cells appeared alone in colonies of less than 50 cells, together with macrophages in mixed colonies and also within clusters of T lymphocytes at high culture cell numbers. The morphologic resemblance to lymphoid dendritic cells was confirmed by electron microscopy and the cells were distinguished from macrophages by immunoenzymatic and immunogold labeling with monoclonal antibodies (MoAbs). Like macrophages they were HLA-DR+ and CD4+. However, they lacked nonspecific esterase and the macrophage cytoplasmic marker Y1/82A. Most strikingly, cells were strongly HLA-DQ+ and expressed CD1a (T6), which is characteristic of skin Langerhans cells. Their functional similarity to lymphoid dendritic cells was demonstrated by their ability to stimulate allogeneic mixed leukocyte reactions. Dendritic cell colony numbers were estimated in both bone marrow and peripheral blood of controls and in leukemia and lymphoma patients before and after chemotherapy. Colony numbers were low in control blood and in patients before treatment (less than 1.0 to 3.7/10(5) cells). However, during hematopoietic recovery the mean value increased to 37.5/10(5) cells and this increase correlated closely with the observed increase in circulating colony forming unit-granulocyte macrophage (CFU- GM) in individual patients. Autoradiographic studies demonstrated mitotic activity within CD1a+ colonies and a linear relationship between cultured cells and both pure and mixed colonies was consistent with their derivation from a single precursor. These data indicate that a novel hematopoietic progenitor of dendritic/Langerhans cells (DL-CFU) may now be identified in a clonal assay system and suggest a probable common progenitor for these cells and macrophages.

scholarly journals The Effect of Influenza Vaccines on Maturation of Dendritic Cells Generated from Bone Marrow

2021 ◽  
Vol 5 (1) ◽  
Author(s):  
Kostinova AM ◽  
◽  
Yukhacheva DV ◽  
Akhmatova EA ◽  
Akhmatova NK ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Dendritic Cells ◽  
Bone Marrow Cells ◽  
Human Bone ◽  
Influenza Vaccines ◽  
Human Bone Marrow ◽  
Vitro Model ◽  
Cytokine Stimulation ◽  
Marrow Cells

Background: Possibility to control immune system by regulating the activity of Dendritic Cells (DC) with the help of vaccines or other immunobiological drugs opens great prospects for infectious, oncological and autoimmune control. The aim of this study was to evaluate in vitro the effect of adjuvant subunit and non-adjuvant split influenza vaccines on maturation of DCs from human bone marrow. Methods: From bone marrow cells of healthy volunteers, DCs were obtained using rGM-CSF and IL-4. On the 8th day of cultivation, 10μl of vaccines against influenza were introduced into the culture of Immature DCs (i-DCs): a non-adjuvant split vaccine (Vaxigripp, Sanofi Pasteur) and an immunoadjuvant subunit vaccine (Grippol plus, Petrovax), as well as immunomodulator Polyoxidonium. Results: Insertion of influenza vaccines into i-DC culture induced the acquisition by DCs typical morphological signs of maturation. DCs became large with eccentrically located of irregular shape nucleus, densified cytoplasm, numerous processes. By immunophenotypic examination decrease in monocyte/macrophage pool, cells with expression of CD34 immaturity marker, increase in expressing CD11c/CD86 costimulatory molecules and CD83 terminal differentiation molecules were observed. Although Polyoxidonium caused a decrease in number of CD11c/CD14 cells (18, 5%), but compared to vaccines, its activity was lower (p<0, 05). Grippol plus more actively induced differentiation of TLR2 and TLR8 expressing cells, whereas Vaxigripp-expression of TLR4 and TLR8 on DCs. Conclusion: The possibility of using in vitro model of DCs obtained from human bone marrow cells by cytokine stimulation for examination of the ability of influenza vaccines to induce DC maturation processes has been demonstrated.

scholarly journals Expression of the YB5.B8 antigen (c-kit proto-oncogene product) in normal human bone marrow

Blood ◽  
1991 ◽  
Vol 78 (1) ◽  
pp. 30-37 ◽  
Author(s):  
LK Ashman ◽  
AC Cambareri ◽  
LB To ◽  
RJ Levinsky ◽  
CA Juttner
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Bone Marrow Cells ◽  
Lymphoid Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Hematopoietic Stem ◽  
Normal Peripheral Blood ◽  
Normal Human ◽  
Normal Human Bone Marrow

Abstract The c-kit proto-oncogene product is a member of the family of growth factor receptors with intrinsic tyrosine kinase activity. In the mouse c-kit maps to the W locus, which is known to be of central importance in hematopoiesis. Monoclonal antibody (MoAb) YB5.B8, which was raised against peripheral blood blast cells from a patient with acute myeloid leukemia (AML), was recently shown to bind to the extracellular domain of the c-kit product. This antibody does not bind detectably to normal peripheral blood cells and identifies a sub-group of AML patients with poor prognosis. We have used MoAb YB5.B8 to study the expression of c- kit by normal human bone marrow cells by immunofluorescence and flow cytometry, and to isolate multipotential and erythroid colony-forming cells. In a series of 11 normal adult bone marrow specimens, MoAb YB5.B8 bound to 4.0% +/- 1.8% of the cells in the low-density fraction. Dual-labeling experiments were performed with YB5.B8, and CD33, CD34, and CD10 MoAbs. Three populations of cells binding YB5.B8 could be identified based on their pattern of coexpression of the other markers; ie, YB5.B8+/CD34+/CD33-, YB5.B8+/CD34+/CD33+ and YB5.B8+/CD34+/CD33+. These populations had distinctive two-dimensional light scatter characteristics and are likely to correspond to precursor colony- forming cells, colony-forming cells, and maturing mast cells, respectively. No cells binding both YB5.B8 and an MoAb to the early lymphoid marker CD10 were found, implying that most early lymphoid cells do not express c-kit. MoAbs to the c-kit protein should prove valuable in multimarker studies of human hematopoietic stem and progenitor cells. Definition of a reference range of c-kit expression in normal human bone marrow will provide a sound basis for further studies of this marker in diagnosis and prognosis in AML.

Characterisation of Side Population (SP) Cells Obtained from Different Hematopoietic Progenitor/Stem Cell Compartments in Human Bone Marrow and Peripheral Blood.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4170-4170
Author(s):  
Dag Josefsen ◽  
Lise Forfang ◽  
Marianne Dyrhaug ◽  
Gunnar Kvalheim
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Cell Population ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Hematopoietic Stem ◽  
Cell Compartments ◽  
Cd34 Cells

Abstract Side population (SP) cells are characterised by their ability to exclude Hoechst 33342 dye from the cells. Using this method, it has been demonstrated that cells within the SP+ fraction of mononuclear cells from both murine and human hematopoietic systems are enriched for primitive hematopoietic stem- and progenitor cells. Moreover, most of the SP+ cells did not express CD34, indicating the presence of a CD34 negative hematopoietic stem cell population. To explore this further, we have examined SP+ cells obtained from different cell compartments in human bone marrow and peripheral blood. Human bone marrow (BM) was obtained from healthy volunteer donors by iliac crest aspiration after informed consent. Mononuclear cells (MNC) were obtained by Ficoll grade centrifugation. CD34+ cells were then isolated from MNC. Highly enriched CD34+ cells were isolated from PBPC obtained from patients with Hodgkin lymphoma. To identify the SP+ cells, the cells were stained with Hoechst 33342 dye. Using flowcytometric techniques (FACStar+, FACSDiva, Becton Dickinson, San Jose, CA) we were able to visualize the dye efflux in SP+ cells. SP+ cells were functionally confirmed using Verapamil. Phenotypical characterisation of the different cell populations using flow cytometric methods was performed. The level of SP+ cells in BM-MNC was 1,3% (mean, n=3) In line with previous findings, we observed that SP+ cells obtained from BM-MNC lack expression of several lineage committed markers, including CD15 and CD19. Most of the cells were CD34− (mean=2,2%), which was lower than in the main population (MP; mean=5%). The level of CD133 expression was low and similar in both populations. Furthermore we found a higher fraction of CD3+ T-cells in the SP fraction than in the MP fraction (mean: 69% vs 51%). To further investigate the SP+CD34+ cell fraction, we examined CD34+ cells isolated from both human bone marrow and peripheral blood. The percentage of SP+CD34+ cells varied from 0,4 up to 18% of the total CD34+ cell population obtained from PBPC (n= 16), whereas the level of SP+CD34+ cells obtained from bone marrow was 5% of the total CD34+ cell population (n=3). Expression of lineage committed markers, including CD10, CD15 and CD19 was less then 10% of the whole CD34+ cell population obtained from PBPC, whereas we found a higher level of expression of these markers in CD34+ cells isolated from bone marrow. However, when we examined the SP+CD34+ cells from either PBPC or bone marrow, we observed that the phenotypic profile of these cells were similar with almost no expression of lineage markers. The frequency of LTC-IC was markedly increased in SP+MNC, in line with previous findings. In addition we also observed a marked increase in LTC-IC in SP+CD34+ cells compared to SP-CD34+ cells in both BM and PB (BM: 7-fold increase; PB: 3–4 fold). In conclusion, SP cells are present in different hematopoietic progenitor cell populations, including BM-MNC, BM-CD34+ cells and PB-CD34+ cells. In SP+CD34+ cell fractions from both BM and PB we observed an increased expression of stem cell markers like CD90 and CD133, whereas in SP+MNC we found low levels of CD34, CD90 and CD133 expression. However, the LTC-IC frequency was markedly higher in all SP+fractions compared to MP fractions, suggesting that sorting of SP+ cells from different hematopoietic stem- and progenitor cell compartments identify immature hematopoietic cells.

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 ◽  
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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