scholarly journals Human fetal liver MSCs are more effective than adult bone marrow MSCs for their immunosuppressive, immunomodulatory, and Foxp3+ T reg induction capacity

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yi Yu ◽  
Alejandra Vargas Valderrama ◽  
Zhongchao Han ◽  
Georges Uzan ◽  
Sina Naserian ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Immune Responses ◽  
Molecular Mechanisms ◽  
Fetal Liver ◽  
Cytotoxic T Cells ◽  
Cell Contact ◽  
Adult Bone Marrow ◽  
Adult Bone

Abstract Background Mesenchymal stem cells (MSCs) exhibit active abilities to suppress or modulate deleterious immune responses by various molecular mechanisms. These cells are the subject of major translational efforts as cellular therapies for immune-related diseases and transplantations. Plenty of preclinical studies and clinical trials employing MSCs have shown promising safety and efficacy outcomes and also shed light on the modifications in the frequency and function of regulatory T cells (T regs). Nevertheless, the mechanisms underlying these observations are not well known. Direct cell contact, soluble factor production, and turning antigen-presenting cells into tolerogenic phenotypes, have been proposed to be among possible mechanisms by which MSCs produce an immunomodulatory environment for T reg expansion and activity. We and others demonstrated that adult bone marrow (BM)-MSCs suppress adaptive immune responses directly by inhibiting the proliferation of CD4+ helper and CD8+ cytotoxic T cells but also indirectly through the induction of T regs. In parallel, we demonstrated that fetal liver (FL)-MSCs demonstrates much longer-lasting immunomodulatory properties compared to BM-MSCs, by inhibiting directly the proliferation and activation of CD4+ and CD8+ T cells. Therefore, we investigated if FL-MSCs exert their strong immunosuppressive effect also indirectly through induction of T regs. Methods MSCs were obtained from FL and adult BM and characterized according to their surface antigen expression, their multilineage differentiation, and their proliferation potential. Using different in vitro combinations, we performed co-cultures of FL- or BM-MSCs and murine CD3+CD25−T cells to investigate immunosuppressive effects of MSCs on T cells and to quantify their capacity to induce functional T regs. Results We demonstrated that although both types of MSC display similar cell surface phenotypic profile and differentiation capacity, FL-MSCs have significantly higher proliferative capacity and ability to suppress both CD4+ and CD8+ murine T cell proliferation and to modulate them towards less active phenotypes than adult BM-MSCs. Moreover, their substantial suppressive effect was associated with an outstanding increase of functional CD4+CD25+Foxp3+ T regs compared to BM-MSCs. Conclusions These results highlight the immunosuppressive activity of FL-MSCs on T cells and show for the first time that one of the main immunoregulatory mechanisms of FL-MSCs passes through active and functional T reg induction.

scholarly journals Human fetal liver MSCs are more effective than adult bone marrow MSCs for their immunosuppressive, immunomodulatory and Foxp3+ T regs induction capacity

2020 ◽  
Author(s):  
Yi Yu ◽  
Alejandra Vargas Valderrama ◽  
Zhongchao Han ◽  
Georges Uzan ◽  
Sina Naserian ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Immune Responses ◽  
Molecular Mechanisms ◽  
Fetal Liver ◽  
Cell Contact ◽  
Adult Bone Marrow ◽  
Wide Range ◽  
Adult Bone

Abstract Background: Mesenchymal stem cells (MSCs) display active capacities of suppressing or modulating harmful immune responses through diverse molecular mechanisms. These cells are under extensive translational efforts as cell therapies for immune-mediated diseases and transplantations. A wide range of preclinical studies and limited number of clinical trials using MSCs have not only shown promising safety and efficacy profiles but have also revealed changes in regulatory T cell (T reg) frequency and function. However, the mechanisms underlying this important observation are not well understood. Cell-to-cell contact, production of soluble factors, reprogramming of antigen presenting cells to tolerogenic phenotypes have emerged as possible mechanisms by which MSCs produce an immunomodulatory environment for T reg expansion. We and others demonstrated that adult bone-marrow (BM)-MSCs suppress adaptive immune responses directly by inhibiting the proliferation of CD4+ (“helper”) and CD8+ (“cytotoxic”) T cells but also indirectly through induction of Tregs. In parallel we demonstrated that fetal liver (FL)-MSCs displays much longer-lasting immunomodulatory properties compared to BM-MSCs, by inhibiting directly the proliferation and activation of CD4+ and CD8+ T cells. Therefore, we investigated if FL-MSCs exert their strong immunosuppressive effect also indirectly through induction of T regs.Methods: MSCs were obtained from FL and adult BM and characterized according to their surface antigen expression, their multilineage differentiation and their proliferation potential. Using different in-vitro combinations, we performed co-cultures of FL or BM-MSCs and murine CD3+CD25-T cells to investigate immunosuppressive effects of MSCs on T cells and to quantify their capacity to induce functional T regs. Results: We demonstrated that although both types of MSC exhibit similar phenotypic profile and differentiation capacity, FL-MSCs have significantly higher proliferative capacity and ability to suppress both CD4+ and CD8+ murine T cell proliferation and to modulate them towards less active phenotypes than adult BM-MSCs. Moreover, their substantial suppressive effect was associated with an outstanding increase of functional CD4+CD25+Foxp3+ T regs compared to BM-MSCs.Conclusions: These results highlight the immunosuppressive activity of FL-MSCs on T cells and show for the first time that one of the main immunoregulatory mechanisms of FL-MSCs passes through active and functional T reg induction.

scholarly journals Human Fetal Liver MSCs are More Effective Than Adult Bone Marrow MSCs for Their Immunosuppressive, Immunomodulatory and Foxp3+ T regs Induction Capacity

2020 ◽  
Author(s):  
Yi Yu ◽  
Alejandra Vargas Valderrama ◽  
Zhongchao Han ◽  
Georges Uzan ◽  
Sina Naserian ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
T Cell ◽  
Immune Responses ◽  
Molecular Mechanisms ◽  
Fetal Liver ◽  
Cell Contact ◽  
Adult Bone Marrow ◽  
Wide Range ◽  
Adult Bone

Abstract Background: Mesenchymal stem cells (MSCs) display active capacities of suppressing or modulating harmful immune responses through diverse molecular mechanisms. These cells are under extensive translational efforts as cell therapies for immune-mediated diseases and transplantations. A wide range of preclinical studies and limited number of clinical trials using MSCs have not only shown promising safety and efficacy profiles but have also revealed changes in regulatory T cell (T reg) frequency and function. However, the mechanisms underlying this important observation are not well understood. Cell-to-cell contact, production of soluble factors, reprogramming of antigen presenting cells to tolerogenic phenotypes have emerged as possible mechanisms by which MSCs produce an immunomodulatory environment for T reg expansion. We and others demonstrated that adult bone-marrow (BM)-MSCs suppress adaptive immune responses directly by inhibiting the proliferation of CD4+ (“helper”) and CD8+ (“cytotoxic”) T cells but also indirectly through induction of Tregs. In parallel we demonstrated that fetal liver (FL)-MSCs displays much longer-lasting immunomodulatory properties compared to BM-MSCs, by inhibiting directly the proliferation and activation of CD4+ and CD8+ T cells. Therefore, we investigated if FL-MSCs exert their strong immunosuppressive effect also indirectly through induction of T regs.Methods: MSCs were obtained from FL and adult BM and characterized according to their surface antigen expression, their multilineage differentiation and their proliferation potential. Using different in-vitro combinations, we performed co-cultures of FL or BM-MSCs and murine CD3+CD25-T cells to investigate immunosuppressive effects of MSCs on T cells and to quantify their capacity to induce functional T regs. Results: We demonstrated that although both types of MSC exhibit similar phenotype profile and differentiation capacity, FL-MSCs have significantly higher proliferative capacity and ability to suppress both CD4+ and CD8+ murine T cell proliferation and to modulate them towards less active phenotypes than adult BM-MSCs. Moreover, their substantial suppressive effect was associated with an outstanding increase of functional CD4+CD25+Foxp3+ T regs compared to BM-MSCs.Conclusions: These results highlight the immunosuppressive activity of FL-MSCs on T cells and show for the first time that one of the main immunoregulatory mechanisms of FL-MSCs passes through active and functional T reg induction.

HCA, an Immunoglobulin-Like Adhesion Molecule Present on the Earliest Human Hematopoietic Precursor Cells, Is Also Expressed by Stromal Cells in Blood-Forming Tissues

Blood ◽  
1999 ◽  
Vol 93 (3) ◽  
pp. 826-837 ◽  
Author(s):  
Fernando Cortés ◽  
Frédéric Deschaseaux ◽  
Nobuko Uchida ◽  
Marie-Claude Labastie ◽  
Annabelle M. Friera ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Fetal Liver ◽  
Present Report ◽  
Marrow Stromal Cells ◽  
Lymphoid Tissues ◽  
Adult Bone Marrow ◽  
Adhesive Interactions ◽  
Adult Bone

We have previously shown that the HCA/ALCAM (CD166) glycoprotein, a member of the immunoglobulin family that mediates both homophilic and heterophilic cell-cell adhesion, via the CD6 ligand, is expressed at the surface of all of the most primitive CD38−/lo, Thy-1+, rho123lo, CD34+hematopoietic cells in human fetal liver and fetal and adult bone marrow. In the present report we show that HCA is also expressed by subsets of stromal cells in the primary hematopoietic sites that sequentially develop in the human embryo and fetus, ie, the paraaortic mesoderm, liver, thymus, and bone marrow. Adult bone marrow stromal cells established in vitro, including those derived from Stro-1+ progenitors and cells from immortalized cell lines, express HCA. In contrast, no HCA expression could be detected in peripheral lymphoid tissues, fetal spleen, and lymph nodes. HCA membrane molecules purified from marrow stromal cells interact with intact marrow stromal cells, CD34+ CD38−hematopoietic precursors, and CD3+ CD6+peripheral blood lymphocytes. Finally, low but significant levels of CD6 are here for the first time detected at the surface of CD34+ rho123med/lo progenitors in the bone marrow and in mobilized blood from healthy individuals. Altogether, these results indicate that the HCA/ALCAM surface molecule is involved in homophilic or heterophilic (with CD6) adhesive interactions between early hematopoietic progenitors and associated stromal cells in primary blood-forming organs.

HCA, an Immunoglobulin-Like Adhesion Molecule Present on the Earliest Human Hematopoietic Precursor Cells, Is Also Expressed by Stromal Cells in Blood-Forming Tissues

Blood ◽  
1999 ◽  
Vol 93 (3) ◽  
pp. 826-837 ◽  
Author(s):  
Fernando Cortés ◽  
Frédéric Deschaseaux ◽  
Nobuko Uchida ◽  
Marie-Claude Labastie ◽  
Annabelle M. Friera ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Fetal Liver ◽  
Present Report ◽  
Marrow Stromal Cells ◽  
Lymphoid Tissues ◽  
Adult Bone Marrow ◽  
Adhesive Interactions ◽  
Adult Bone

Abstract We have previously shown that the HCA/ALCAM (CD166) glycoprotein, a member of the immunoglobulin family that mediates both homophilic and heterophilic cell-cell adhesion, via the CD6 ligand, is expressed at the surface of all of the most primitive CD38−/lo, Thy-1+, rho123lo, CD34+hematopoietic cells in human fetal liver and fetal and adult bone marrow. In the present report we show that HCA is also expressed by subsets of stromal cells in the primary hematopoietic sites that sequentially develop in the human embryo and fetus, ie, the paraaortic mesoderm, liver, thymus, and bone marrow. Adult bone marrow stromal cells established in vitro, including those derived from Stro-1+ progenitors and cells from immortalized cell lines, express HCA. In contrast, no HCA expression could be detected in peripheral lymphoid tissues, fetal spleen, and lymph nodes. HCA membrane molecules purified from marrow stromal cells interact with intact marrow stromal cells, CD34+ CD38−hematopoietic precursors, and CD3+ CD6+peripheral blood lymphocytes. Finally, low but significant levels of CD6 are here for the first time detected at the surface of CD34+ rho123med/lo progenitors in the bone marrow and in mobilized blood from healthy individuals. Altogether, these results indicate that the HCA/ALCAM surface molecule is involved in homophilic or heterophilic (with CD6) adhesive interactions between early hematopoietic progenitors and associated stromal cells in primary blood-forming organs.

Efficient Expression of a Kit/GFP Transgene in Hematopoietic Stem Cells of Mouse Fetal Liver and Bone Marrow.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4159-4159
Author(s):  
Francesco Cerisoli ◽  
Letizia Cassinelli ◽  
Giuseppe Lamorte ◽  
Stefania Citterio ◽  
Maria Cristina Magli ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Fetal Liver ◽  
Hematopoietic Progenitors ◽  
Hematopoietic Stem ◽  
Adult Bone Marrow ◽  
Gfp Fluorescence ◽  
Adult Bone

Abstract The hierarchy of transcription factors and signalling molecules involved in hematopoietic development has been dissected through transgenic and knock-out experiments, leading to the identification of several important genes. Less well known are the networks of transcription factors which regulate the activities of the main genes identified. Kit, encoding the membrane receptor of Stem Cell Factor (SCF), is a critical molecule for Hematopoietic Stem Cells (HSC) and some early progenitors, in which it is expressed. In a previous work (Cairns et al., Blood102, 3954;2003), we used mouse lines expressing transgenic Green Fluorescent Protein (GFP) under the control of Kit regulatory elements to investigate Kit regulation in different cell systems such as the hematopoietic and germ cell lineages. We generated a mouse Kit transgene capable of efficiently driving GFP expression both in PGC and in hematopoietic progenitors, such as CFU-Mix and BFU-Es. In the present work, we evaluated the functional efficiency of the same transgene also in HSC residing in the Fetal Liver (FL) and adult Bone Marrow (BM). To test if the construct is expressed in HSC, we transplanted FL or BM cells, fractionated on the basis of Kit expression and the level of GFP fluorescence, into irradiated non-transgenic mice. At the same time, the proportion of hematopoietic progenitors in the various fractions was assessed by in vitro colony assays. Following long term hematological reconstitution, the contribution of transplanted GFP cells was evaluated by the proportion of fluorescent mixed colonies in colture as well as by the proportion of fluorescent bone marrow cells, as assessed by FACS analysis. Long term reconstitution was confirmed by secondary transplants. Results show that the repopulating cells derived from fetal liver and adult bone marrow reside in a fraction of Kit+ cells with intermediate GFP fluorescence level, whereas CFU-Mix and BFU-E are in the highly GFP fluorescent fraction. Furthermore, flow cytometry of fetal liver shows that the intermediate fluorescence fraction is highly enriched in Kit+, Sca1+, CD11b+ cells (the expected HSC immunophenotype), whereas the high fluorescence fraction contains mainly Kit+, Sca1−, CD11b− cells. Similarly, the HSC-enriched tip of the Side Population (SP) of adult bone marrow is highly enriched in Kit+, Sca1+ cells of intermediate GFP fluorescence, whereas the upper part of the SP is enriched in Kit+, Sca1− cells of high GFP fluorescence. Our results indicate that the transgene (and possibly the endogenous Kit gene as well) might be transcribed at relatively low levels in HSC versus other progenitors. Noteworthy, the same transgene is also highly expressed in PGC and in Cardiac Stem Cells (CSC) (Messina et al., Circ. Res. 95,911;2004) and in blastocyst inner mass grown in vitro, indicating that the most 5′ part of the intron (4kb), added to the otherwise inactive promoter might include sites regulating Kit expression in multiple stem cell types.

scholarly journals The Translocation t(7;12)(q36;p13) Induces Myeloid Leukemia in Immuno-Compromised but Not Immunocompetent Mice

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 2707-2707
Author(s):  
Ahmed Waraky ◽  
Anders Östlund ◽  
Laleh Arabanian ◽  
Tina Nilsson ◽  
Linda Fogelstrand ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Fetal Liver ◽  
Liver Cells ◽  
Adult Bone Marrow ◽  
Leukemia Development ◽  
Fetal Liver Cells ◽  
Immunocompetent Mice ◽  
Adult Bone

Introduction: Non-random cytogenetic aberrations are often involved in the development of AML in children and several aberrations can serve as diagnostic markers, prognosis predictors and impact choice of therapy. In infant AML, a chromosomal translocation t(7;12)(q36;p13) has been found in up to 20-30 % of the cases, making it the second most common genetic aberration in this age group after KMT2A (MLL) rearrangements. Previous studies indicate that this patient group has a dismal prognosis with virtually no event-free survival. Limiting the chances to improve this is the lack of understanding how the t(7;12)(q36;p13) is involved in leukemia development. The translocation leads to a gene fusion MNX1-ETV6 but also to increased MNX1 gene expression. Although both ETV6 and MNX1 are expressed in normal hematopoietic tissues, the role of the fusion protein MNX1-ETV6in the development of AML is not established. Also unclear is whether the driver of leukemogenesis is the fusion itself or the simultaneous overexpression of MNX1. The aim of this study was to assess the transformation capacity and the molecular mechanism of the MNX1-ETV6 fusion and the overexpressed MNX1in vitro and in vivo using murine transplantation models. Material and methods: In a liquid culture system, we introduced the MNX1-ETV6 fusion, MNX1 overexpression, or empty vector into primary murine (C57BL/6) hematopoietic progenitor cells with retroviral transfection. Cells were isolated from either adult bone marrow after 5-FU stimulation, or from fetal liver at E14.5. After enrichment by fluorescence activated cell sorting based on vector co-expressed green/yellow fluorescence protein, transfected cells were used for in vitro experiments and for transplantation into lethally irradiated immunocompetent C57BL/6 mice or sub-lethally irradiated immunocompromised NSGW41 mice. In vitro, cells were assessed with RNA sequencing for gene expression, gamma H2AX assay for DNA double strand brakes, flow cytometry for lineage marker expression, apoptosis and proliferation, and with colony forming unit assay. Results: Upon transplantation, only fetal liver cells transduced with MNX1 or with MNX1-ETV6 fusion were able to induce leukemia in immunocompromised (NSGW41) mice. When MNX1 or MNX1-ETV6 transduced cells were transplanted into immunocompetent mice (C57BL/6) mice, no leukemia development was seen, when either fetal liver or adult bone marrow cells were used for transduction. However, when immunocompromised mice were transplanted with MNX1 or MNX1-ETV6 fusion expressing cells they typically developed signs of disease after 1-2 months and exhibited leukocytosis and elevated blast cells in blood and bone marrow, severe anemia, and enlarged spleens with infiltration of leukemic cells. The cells showed expression of predominantly myeloid markers. In vitro, cells with overexpression of MNX1 or MNX1-ETV6 fusion expression also showed altered lineage differentiation in favor of myeloid differentiation. In addition, MNX1 overexpressing cells, but not MNX1-ETV6 expressing cells, exhibited increased proliferation and colony formation capacity. Both MNX1 overexpressing and MNX1-ETV6 fusion expressing cells showed increased DNA damage as evident from an increased gamma-phosphorylated H2AX in fetal liver and adult bone marrow transduced cells respectively, accompanied with G1 arrest, compared to cells transduced with empty vectors. Both MNX1 and MNX1-ETV6 also led to increased apoptosis in adult bone marrow (3-fold) and to a lesser extent in fetal liver cells (1.5-fold). Results from transcriptome sequencing showed enrichment for specific pathways in G2/M transition of cell cycle in cells transduced by either MNX1or the MNX1-ETV6 fusion. Further investigations to elucidate the molecular mechanisms and pathways through which MNX1 and/or MNX1-ETV6 induce leukemia is ongoing. Conclusions: MNX1 overexpression and MNX1-ETV6 fusion, both characteristics of infant AML with t(7;12)(q36;p13), induced leukemogenic effects in both fetal liver cells and adult bone marrow cells, but could cause a myeloid leukemia only under immunocompromised conditions. This may be of importance for the exclusive prevalence of this AML subtype in young children, with the highest peak during the first six months of life when the immune system is less developed. Disclosures No relevant conflicts of interest to declare.

T Cell Progenitors in the Murine Fetal Liver: Differences from Those in the Adult Bone Marrow

1997 ◽  
Vol 177 (1) ◽  
pp. 18-25 ◽  
Author(s):  
Yoshihiro Watanabe ◽  
Yuichi Aiba ◽  
Yoshimoto Katsura
Keyword(s):  
Bone Marrow ◽  
T Cell ◽  
Fetal Liver ◽  
Adult Bone Marrow ◽  
T Cell Progenitors ◽  
Adult Bone
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