Suppressor of Cytokine Signaling 3 Promotes Bone Marrow Cells to Differentiate into CD8+ T Lymphocytes in Lung Tissue via Up-Regulating Notch1 Expression

Abstract Abstract 4870 Background and objectives: Human leukocyte antigen –G (HLA–G) is a nonclassic HLA class I molecule and has the immunosuppressive activity. We investigate the effects of Granulocyte colony-stimulating factor (G-CSF) on the expression and secretion level of HLA-G in peripheral blood and bone marrow cells, and attempt to explain the mechanism of the low incidence of graft-versus-host disease (GVHD) in G-CSF-primed bone marrow (G-BM) or peripheral blood stem cells (G-PBSC) transplantation. Methods: Bone marrow and peripheral blood cells of 10 donors were collected pre-mobilization and at day 5 after G-CSF mobilization. Flow cytometry was used to detect the expression of membrane-bound HLA-G (mHLA-G) of CD3+CD4+ T lymphocytes, CD3+CD8+ T lymphocytes, CD19+ B lymphocytes, CD56+ NK cell, CD14+ monocytes and CD33+ granulocytes. The levels of soluble HLA-G (sHLA-G) and cytokine were determined by enzyme-linked immunosorbent assay (ELISA). Bone marrow cells and peripheral blood cells of pre-mobilization were incubated for 24h, which are divided into 5 groups: (a) blank group, (b) with G-CSF, (c) with G-CSF and blocking antibody of IL-10, (d) with G-CSF and blocking antibody of IFN-γ, (e) with G-CSF, blocking antibody of IL-10 and blocking antibody of IFN-γ. Results: The levels of mHLA-G of CD3+CD4+ lymphocytes, CD3+CD8+ lymphocytes and the levels of sHLA-G at day 5 after G-CSF mobilization groups were significantly higher than the pre-mobilization groups both in bone marrow and peripheral blood cells, especially in CD3+CD8+ lymphocytes. The levels of mHLA-G of CD19+ B lymphocytes, CD56+ NK cells, CD14+ monocytes and CD33+ granulocytes had no significant difference in the two groups. The levels of IL-10 or IFN-γ in plasma of bone marrow or peripheral blood at day 5 after G-CSF mobilization groups were much higher than the pre-mobilization groups. The levels of mHLA-G of CD3+CD4+ lymphocytes, CD3+CD8+ lymphocytes and the levels of sHLA-G in bone marrow cells or peripheral blood cells incubating with G-CSF for 24h were significantly higher than the groups without incubating with G-CSF. And the blocking antibody of IL-10 or the blocking antibody of IFN-γ could not reduce the expression levels of mHLA-G and sHLA-G in vitro. Conclusion: G-CSF can act on cells directly to increase the expression levels of HLA-G in CD3+CD4+ T lymphocytes and CD3+CD8+ T lymphocytes. It might be assciated with the low incidence of GVHD in G-BM transplantation or G-PBSC transplantation. Supported by National Natural Science Foundation of China (30971300), Science and Technology Planning Project of Guangdong Province of China (2009A030200007) and China Postdoctoral Science Foundation (200902332, 20080440776). Disclosures: No relevant conflicts of interest to declare.

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The identification in adult bone marrow of pluripotent and restricted stem cells of the myeloid and lymphoid systems

Journal of Experimental Medicine ◽

10.1084/jem.145.6.1567 ◽

1977 ◽

Vol 145 (6) ◽

pp. 1567-1579 ◽

Cited By ~ 348

Author(s):

S Abramson ◽

RG Miller ◽

RA Phillips

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Stem Cell ◽

T Lymphocytes ◽

Chromosome Aberrations ◽

Bone Marrow Cells ◽

Hemopoietic Stem Cell ◽

Lymphoid System ◽

Unique Chromosome ◽

Marrow Cells

The precise relationship between the stem cells for the lymphoid system and those for the blood-forming system is unclear. While it is generally assumed that the hemopoietic stem cell, the spleen colony-forming unit (CFU-S), is also the stem cell for the lymphoid system, there is little evidence for this hypothesis. To investigate the stem cells in these two systems, we irradiated bone marrow cells to induce unique chromosome aberrations in the stem cell population and injected them at limiting dilution into stem cell-deficient recipients. Several months (between 3 and 11) were allowed for the injected cells to repopulate the hemopoietic system. At that time, the bone marrow, spleen, and thymus were examined for a high frequency of cells having the same unique chromosome aberration. The presence of such markers shows that the marker was induced in a cell with extensive proliferative capacity, i.e., a stem cell. In addition, the splenic lymphocytes were stimulated with phytohemagglutinin (PHA) or lipopolysaccharide (LPS) to search for unique chromosomes in dividing T and B cells, respectively. Finally, bone marrow cells were injected into secondary irradiated recipients to determine if the marker occurred in CFU-S and to determine whether or not the same tissue distributions of marked cells could be propogated by bone marrow cells in a second recipient. After examination of 28 primary recipients, it was possible to identify three unique patterns of stem cell regeneration. In one set of mice, a unique chromosome marker was observed in CFU-S and in PHA- and LPS-stimulated cultures. These mice provide direct evidence for a pluripotent stem cell in bone marrow. In addition, two restricted stem cells were identified by this analysis. In three recipients, abnormal karyotypes were found only in myeloid cells and not in B and T lymphocytes. These mice presumably received a marked stem cell restricted to differentiate only into myeloid progeny. In three other recipients, chromosome aberrations were found only in PHA-stimulated cells; CFU-S and cells from LPS cultures did not have cells with the unique chromosome. This pattern suggests that bone marrow contains cells committed to differentiation only into T lymphocytes. For each of the three types of stem cells, secondary recipients had the same cellular distribution of marked cells as the primary recipients. This observation provides further evidence that unique markers can be induced in both pluripotent and restricted stem cells.

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Extrathymic Development and Function of Human T-Lymphocytes from Bone Marrow Cells in Vitro

Cellular Immunology ◽

10.1006/cimm.1994.1054 ◽

1994 ◽

Vol 154 (1) ◽

pp. 25-42 ◽

Cited By ~ 8

Author(s):

M. Adibzadeh ◽

H.J. Bühring ◽

T. Daikeler ◽

P. Siegels-Hübenthal ◽

M. Owsianowsky ◽

...

Keyword(s):

Bone Marrow ◽

T Lymphocytes ◽

Bone Marrow Cells ◽

Human T Lymphocytes ◽

And Function ◽

Marrow Cells

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Abnormal hematopoiesis in Gab2 mutant mice

Blood ◽

10.1182/blood-2006-11-060707 ◽

2007 ◽

Vol 110 (1) ◽

pp. 116-124 ◽

Cited By ~ 36

Author(s):

Yi Zhang ◽

Ernesto Diaz-Flores ◽

Geqiang Li ◽

Zhengqi Wang ◽

Zizhen Kang ◽

...

Keyword(s):

Bone Marrow ◽

Bone Marrow Cells ◽

Mapk Signaling ◽

Cytokine Signaling ◽

Normal Numbers ◽

Hematopoietic Growth Factors ◽

Extracellular Signal ◽

Fold Reduction ◽

Marrow Cells

Gab2 is an important adapter molecule for cytokine signaling. Despite its major role in signaling by receptors associated with hematopoiesis, the role of Gab2 in hematopoiesis has not been addressed. We report that despite normal numbers of peripheral blood cells, bone marrow cells, and c-Kit+Lin−Sca-1+ (KLS) cells, Gab2-deficient hematopoietic cells are deficient in cytokine responsiveness. Significant reductions in the number of colony-forming units in culture (CFU-C) in the presence of limiting cytokine concentrations were observed, and these defects could be completely corrected by retroviral complementation. In earlier hematopoiesis, Gab2-deficient KLS cells isolated in vitro responded poorly to hematopoietic growth factors, resulting in an up to 11-fold reduction in response to a cocktail of stem cell factor, flt3 ligand, and thrombopoietin. Gab2-deficient c-Kit+Lin− cells also demonstrate impaired activation of extracellular signal-regulated kinase (ERK) and S6 in response to IL-3, which supports defects in activating the phosphatidylinositol-3 kinase (PI-3K) and mitogen-associated protein kinase (MAPK) signaling cascades. Associated with the early defects in cytokine response, competitive transplantation of Gab2−/− bone marrow cells resulted in defective long-term multilineage repopulation. Therefore, we demonstrate that Gab2 adapter function is intrinsically required for hematopoietic cell response to early-acting cytokines, resulting in defective hematopoiesis in Gab2-deficient mice.

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Nrf2, a Critical Regulator of Oxidative Stress, Is Required for HSC Function and Cytokine Response.

Blood ◽

10.1182/blood.v114.22.1492.1492 ◽

2009 ◽

Vol 114 (22) ◽

pp. 1492-1492

Author(s):

Akil Merchant ◽

Anju Singh ◽

Giselle Joseph ◽

Qiuju Wang ◽

Ping Zhang ◽

...

Keyword(s):

Oxidative Stress ◽

Stem Cells ◽

Bone Marrow ◽

Transcription Factor ◽

Peripheral Blood ◽

Bone Marrow Cells ◽

Cytokine Signaling ◽

Wild Type ◽

Increased Sensitivity ◽

Marrow Cells

Abstract Abstract 1492 Poster Board I-515 Previous studies have established an important role for reactive oxygen species (ROS) in regulating the function and life-span of hematopoietic stem cells (HSC). Nuclear factor erythroid-2–related factor 2 (Nrf2) is a redox-sensitive transcription factor that regulates cellular responses to ROS and detoxification pathways implicated in chemoresistance, however, its role in normal stem cells is unknown. We analyzed Nrf2null mice and found increased total bone marrow cellularity, cKit+Sca1+Lin− (KSL) stem-progenitor cells, and long-term quiescent HSC (CD34−KSL) compared to wild type mice (p<0.05). Transplantation of equal numbers of KSL cells from Nrf2wt and Nrf2null resulted in a five-fold decrease in peripheral blood chimerism from Nrf2null derived cells at 16 weeks (15% wild type vs. 3% null, p<0.05). Unlike other models of deficiencies in genes associated with ROS handling, such as ATM or the FoxO family of transcription factors, basal ROS levels were not elevated in Nrf2null HSC. However, Nrf2null bone marrow cells demonstrated increased sensitivity to induced oxidative stress and in vitro treatment with H2O2 resulted in a 2 fold decrease in colony formation in methylcellulose. We also examined the in vivo sensitivity of Nrf2null cells to oxidative stress by irradiating (400 rads) stably chimeric mice 20 weeks following transplantation with either Nrf2wt or Nrf2null HSC. Mice receiving Nrf2null HSC demonstrated a 50% decrease in peripheral blood chimerism at 4 months following radiation compared to no change in Nrf2wt recipients (p<0.05) confirming that loss of Nrf2 leads to increased sensitivity to oxidative stress. Microarray gene expression analysis from Nrf2wt and Nrf2null mice revealed down regulation of the G-CSF cytokine receptor in Nrf2null HSC and suggested that defective cytokine signaling may contribute to the HSC dysfunction seen in Nrf2null bone marrow cells. To test this hypothesis, we attempted to rescue the function of Nrf2null HSC by treating mice with exogenous G-CSF. Nrf2wt and Nrf2null mice were treated with one week of daily G-CSF and then HSC were harvested and transplanted. In contrast to the defects in engraftment of untreated Nrf2null HSC, there was no significant difference in peripheral blood chimerism following transplantation of G-CSF treated Nrf2wt or Nrf2null HSC, thus demonstrating that G-CSF treatment could rescue the HSC defect in mutant mice. In conclusion, the Nrf2 transcription factor appears to be a novel and essential regulator of normal HSC function through the modulation of oxidative stress response and cytokine signaling. Disclosures: No relevant conflicts of interest to declare.

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The differentiation of bone marrow cells to functional T lymphocytes following implantation of thymus grafts and thymic stroma in nude and AT×BM mice

Cellular Immunology ◽

10.1016/0008-8749(77)90232-5 ◽

1977 ◽

Vol 34 (1) ◽

pp. 93-103 ◽

Cited By ~ 8

Author(s):

Gary A. Splitter ◽

Travis C. McGuire ◽

William C. Davis

Keyword(s):

Bone Marrow ◽

T Lymphocytes ◽

Bone Marrow Cells ◽

Thymic Stroma ◽

Marrow Cells

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Aleutian Disease Virus in B and T Lymphocytes from Blood and Spleen and in Bone Marrow Cells from Naturally Infected Mink

Intervirology ◽

10.1159/000149553 ◽

1984 ◽

Vol 22 (4) ◽

pp. 211-217 ◽

Cited By ~ 8

Author(s):

Susanne Roth ◽

Oskar-Rüger Kaaden ◽

Sabine van Dawen ◽

Volker Moennig

Keyword(s):

Bone Marrow ◽

T Lymphocytes ◽

Disease Virus ◽

Bone Marrow Cells ◽

Aleutian Disease Virus ◽

B And T Lymphocytes ◽

Marrow Cells

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Helix Pomatia Receptors on Rat T Lymphocytes and Bone Marrow Cells

Immunological Communications ◽

10.3109/08820138209050729 ◽

1982 ◽

Vol 11 (4) ◽

pp. 293-302 ◽

Cited By ~ 1

Author(s):

Joyce A. Killen ◽

James H. Holda ◽

Robert H. Swanborg

Keyword(s):

Bone Marrow ◽

T Lymphocytes ◽

Bone Marrow Cells ◽

Helix Pomatia ◽

Marrow Cells

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Failure in generating hemopoietic stem cells is the primary cause of death from cytomegalovirus disease in the immunocompromised host.

Journal of Experimental Medicine ◽

10.1084/jem.167.5.1645 ◽

1988 ◽

Vol 167 (5) ◽

pp. 1645-1658 ◽

Cited By ~ 66

Author(s):

W Mutter ◽

M J Reddehase ◽

F W Busch ◽

H J Bühring ◽

U H Koszinowski

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

T Lymphocytes ◽

Cause Of Death ◽

Bone Marrow Cells ◽

Immunocompromised Host ◽

Virus Multiplication ◽

Cd8 T Lymphocytes ◽

Cmv Disease ◽

Host Tissues

We have shown in a murine model system for cytomegalovirus (CMV) disease in the immunocompromised host that CMV infection interferes with the earliest detectable step in hemopoiesis, the generation of the stem cell CFU-S-I, and thereby prevents the autoreconstitution of bone marrow after sublethal irradiation. The antihemopoietic effect could not be ascribed to a direct infection of stem cells. The failure in hemopoiesis was prevented by adoptive transfer of antiviral CD8+ T lymphocytes and could be overcome by syngeneic bone marrow transplantation. CD8+ T lymphocytes and bone marrow cells both mediated survival, although only CD8+ T lymphocytes were able to limit virus multiplication in host tissues. We concluded that not the cytopathic effect of virus replication in host tissues, but the failure in hemopoiesis, is the primary cause of death in murine CMV disease.

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