Bone Marrow-Derived Mensenchymal Stem Cells Modified by Mouse Interferon-γ Gene Reduce Fibrosis and Improve Function In a Mouse Model of Liver Fibrosis.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3768-3768
Author(s):  
Hongli Xue ◽  
Shijia Ling ◽  
Dong He ◽  
Yanjun Wen ◽  
Wentong Meng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
Liver Fibrosis ◽  
Adenoviral Vector ◽  
Transfection Efficiency ◽  
Interferon Γ ◽  
Con A ◽  
Ifn Γ ◽  
Mensenchymal Stem Cells

Abstract Abstract 3768 Objective: To investigate the effect of bone marrow-derived mensenchymal stem cells modified by mouse interferon-γ gene on experimental liver fibrosis in mice. Methods: (1) Mesenchymal stem cells (MSCs) derived from BM in BALB/c mice were cultured and identified by the flow cytometry. (2) The Ad-mIFN-γ, a recombinant adenoviral vector encoding mouse IFN-γ gene obtained by PCR from the vector pORF5-mIFN-γ, was created with the AdEasy Adenoviral Vector System. (3) MSCs were transfected with Ad-mIFN-γ. The transfection efficiency was detected by the flow cytometry at 24 hours after infection to determine the best multiplicity of infection (MOI). (4) The liver fibrosis of BALB/c mice were induced with concanavalin A (Con A) challenge weekly for six times. Then MSCs modified by the recombinant adenovirus encoding mouse IFN-γ gene were infused to BALB/c mice induced with Con A at the 24 hours after the first dose. The behavioral changes of mice were monitored closely. One week after the last dose of Con A, all mice were sacrificed and the fibrosis indexes were assessed by several histopathology methods systemicly. Moreover, hepatic hydroxyproline and serum parameters for liver fibrosis and liver function were evaluated and serum mIFN-γ concentrations were measured, respectively. Results: (1) The flow cytometry confirmed that the cells we obtained were MSCs. (2) The target gene mIFN-γ amplified by PCR from the vector pORF5-mIFN-γ, was identified by sequencing, which proved that the mIFN-γ gene was consisted of 468 nucleotides and was completely the same as the sequence published on GenBank. The target gene mIFN-γ was cut out by double endonucleases and then connected to the shuttle vector pAdTrack-CMV. Then the newly constructed vector was linearized by Pme I following transformed to the E.coli. BJ5183, which has the backbone vector of pAdEasy-1. The correct recombinant pAd-mIFN-γ was selected by endonucleases and by Kanamycin resistance and was transfected into AD-293 cells to obtain the adenoviral vector Ad-mIFN-γ. The Ad-mIFN-γ can be propagated in AD-293 cell line, the titre of which was 3.2×109 pfu/ml. (3) MSCs modified by recombinant adenovirus encoding mIFN-γ (IFN-γ/GFP-MSCs) at different multiplicity of infection (MOI) were detected by the flow cytometry. The optimal MOI for Ad-mIFN-γ was 400 with the transfection efficiency of 80.4%, and the optimal MOI for Ad-control was 200 with the transfection efficiency of 72.3%. (4) The changes of liver tissue and serum parameters for liver fibrosis showed liver fibrosis were reducd more obviously in the group treated with IFN-γ/GFP-MSCs (p<0.05), and liver functions were also improved more obviously in this group (p<0.05). The histopathology study using HE staining, masson trichrome staining and silver staining demonstrated that the structure of liver became much better in mice treated with IFN-γ/GFP-MSCs, with liver fibrosis improved significantly. Conclusion: Bone marrow-derived mensenchymal stem cells modified by mouse interferon-γ can reduce liver fibrosis and improve liver function on immuo-mediated liver fibrosis in mice induced by Con A. Disclosures: No relevant conflicts of interest to declare.

Bystander destruction of hematopoietic progenitor and stem cells in a mouse model of infusion-induced bone marrow failure

Blood ◽  
2004 ◽  
Vol 104 (6) ◽  
pp. 1671-1678 ◽  
Author(s):  
Jichun Chen ◽  
Karen Lipovsky ◽  
Felicia M. Ellison ◽  
Rodrigo T. Calado ◽  
Neal S. Young
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Bone Marrow Failure ◽  
Marrow Cell ◽  
Annexin V ◽  
Interferon Γ ◽  
Hematopoietic Progenitor ◽  
Ifn Γ

Abstract Infusion of parental lymph node (LN) cells into sublethally irradiated hybrid F1 recipients created a murine model for bone marrow (BM) failure. Affected animals developed fatal pancytopenia within 2 to 3 weeks, accompanied by BM oligoclonal T-cell infiltration and severe marrow hypoplasia indicated by approximately 10-fold declines in total BM cellularity, 15-fold declines in BM Lin-Sca1+c-Kit+ cells, 100-fold declines in spleen colony-forming units, and 100-fold declines in hematopoietic progenitor and stem cells as estimated by irradiation protection in vivo. LN cells of both H2b/b and H2d/d haplotypes were effectors. Serum interferon-γ (IFN-γ) concentration increased 2- to 3-fold. Marrow cells were severely apoptotic, with high proportions of Fas+ and annexin V+ cells. Cotransplantation of 5 × 105 BM cells from clinically affected donors and 106 BM cells from H2 identical healthy mice could not rescue lethally irradiated recipients. Recipients had significantly lower cellularity in peripheral blood and BM, and cell mixtures failed to produce a stromal feeder layer to support marrow cell growth in vitro. Pathogenic T cells from donors after BM failure appeared capable of destroying hematopoietic progenitor, stem, and stromal cells from fully compatible healthy donors as “innocent bystanders.” This effect can be partially abrogated by anti-IFN-γ antibody. (Blood. 2004;104:1671-1678)

Bone Marrow Stem Cells Contribute to Alcohol Liver Fibrosis in Humans

2010 ◽  
Vol 19 (9) ◽  
pp. 1417-1425 ◽  
Author(s):  
Evangelos Dalakas ◽  
Philip N. Newsome ◽  
Shelagh Boyle ◽  
Rachael Brown ◽  
Anne Pryde ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Liver Fibrosis ◽  
Bone Marrow Stem Cells

scholarly journals Mesenchymal Stem Cells and Induced Bone Marrow-Derived Macrophages Synergistically Improve Liver Fibrosis in Mice

2018 ◽  
Vol 8 (3) ◽  
pp. 271-284 ◽  
Author(s):  
Yusuke Watanabe ◽  
Atsunori Tsuchiya ◽  
Satoshi Seino ◽  
Yuzo Kawata ◽  
Yuichi Kojima ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Liver Fibrosis

scholarly journals Effects of Interferon-γ on Proliferation and Ability of Secretion of Human Amniotic Mesenchymal Stem Cells

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5012-5012
Author(s):  
Ya Gao ◽  
Ying Xu ◽  
Weiru Li ◽  
Yintian Zhang ◽  
Baohong Ping ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Conflicts Of Interest ◽  
Interferon Γ ◽  
Cell Counting ◽  
Inflammatory Factors ◽  
Complete Medium ◽  
Amniotic Mesenchymal Stem Cells ◽  
Soluble Hla ◽  
Ifn Γ

Objective:The immunoregulatory properties and proliferation of mesenchymal stem cells (MSCs) could be affected by inflammatory factors. However, there have been few studies about human amniotic MSCs (hAMSCs). We investigated the effects of interferon (IFN)-γ on the proliferation and apoptosis of hAMSCs, and measured the level of inflammatory factors secreted by hAMSCs. Result:hAMSCs were cultured with complete medium with different concentrations of IFN-γ. We detected the proliferation of hAMSCs by Cell Counting Kit-8 assays, analysed apoptosis by flow cytometry (FCM) at 48 h, and mesasured the level of inflammatory factors such as solube HLA-G and prostaglandin E2 (PGE2) in the supernatant at 48 h by ELISA. The level of kynurenine (KYN) was measured by ultraviolet spectrophotometry. As culture time increased, the proliferation of hAMSCs with different concentrations of IFN-γ increased rapidly from day 1 to day 4, and then the growth rate slowed. FCM indicated that there was no significant apoptosis in the 100 ng/ml IFN-γ group compared with cells without IFN-γ. The level of PGE2 and soluble HLA-G in cells with IFN-γ was higher compared with those without IFN-γ. The level of KYN increased significantly in the cells with IFN-γ. Conclusion:IFN-γ did not affect the growth and proliferation of hAMSCs, and promoted secretion of PGE2 and soluble HLA-G, and enhanced activity of indoleamine 2,3-dioxygenase (IDO), providing a theoretical basis for hAMSCs to prevent and treat graft-versus-host disease. Disclosures No relevant conflicts of interest to declare.

scholarly journals T Cell Subsets and Associated Cytokines in Chronic Graft-Versus-Host-Disease Using G-CSF Primed Bone Marrow in Allogeneic Hematopoietic Stem Cell Transplantation

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 4580-4580
Author(s):  
Monica M Rivera Franco ◽  
Eucario Leon Rodriguez ◽  
Diana Gomez Martin ◽  
Javier Merayo Chalico ◽  
Jorge Alcocer Varela
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Stem Cell ◽  
Th17 Cells ◽  
Mononuclear Cells ◽  
Chronic Gvhd ◽  
The Other ◽  
Hematopoietic Stem ◽  
Graft Versus Host ◽  
Ifn Γ

Abstract Background Graft versus host disease (GVHD) is the major complication of allogeneic hematopoietic stem cell transplantation. It is characterized by an imbalance between the effector and regulatory arms of the immune system which results in the over production of inflammatory cytokines. Regulatory T (T regs) cells and T helper 17 (Th17) cells are two recently described lymphocyte subsets with opposing actions. Both can develop from naïve CD4+ T cell precursors under the influence of TGFβ1. Th17 lymphocytes, are key effector cells in rodent models of human diseases including GVHD. The other subset, T regs, is essential for dominant immunologic tolerance. At our institution, patients transplanted using G-CSF primed bone marrow (G-BM), have a lower incidence of acute and chronic GVHD when compared to those transplanted with peripheral blood and not primed bone marrow. Some microenvironment characteristics of this hematopoietic stem cells (HSC) source remain unknown, as well as the difference between Tregs, Th17 and cytokine levels in patients who develop GVHD and those who do not. Objective To analyze the characteristics of thirty-eight G-BM donor samples, identifying lymphocytes subsets and associated cytokines, and comparing patients who developed chronic GVHD (cGVHD) and those who did not. Materials and Methods A prospective analysis was performed in 38 G-BM samples from donors from 1999 to 2016. Mononuclear cells were defrosted, counted, and viability was evaluated. A 24 hour resting with RPMI, and posterior activation with PMA (50 ng/ml) for 48 hours was performed. Cells were harvested and cytokines were evaluated by flow cytometry (CBA assay). From each sample, one million mononuclear cells were permeabilized, fixed, and stained with CD4-FITC, IL17A-PE, IFN-γ APC, and IL-4 PECy7, for their posterior phenotipication by flow cytometry. The samples were obtained in a BD LSR Fortessa cytometry, and analyzed with the Flow-Jo software. Patients (recipients) information was analyzed using SPSS v.21. Results GVHD incidence was reported as following: Three (8%) patients developed acute GVHD (2 grade II, and 1 grade IV), 11 patients (29%) developed chronic GVHD (9% extensive, and 91% limited), and 24 patients did not present either. Mononuclear cells from G-BM from donors of patients who developed cGVHD showed a pro inflammatory response, characterized by an increased concentration of IL-17A (15.5 vs 0.71 pg/mL, p=0.013), TNF-α (80.27 vs 0.13 pg/mL, p=0.001), and IL-6 (4953.6 vs 11.75 pg/mL, p=0.025), after a mitogenic stimulation, compared to cells from donors of patients who did not developed GVHD. On the other hand, a decreased IL-10 production (2.62 vs 52.81 pg/mL, p=0.001) was documented in mononuclear cells from donors of patients who developed chronic GVHD, compared to donor cells of patients who did not. No significant difference in the production of IL-2, IL-4, and IFN-γ was observed. There was no difference in Th1 and Th2 between both groups, but mononuclear cells from donors of patients who developed chronic GVHD had a higher percentage of Th17 (1.02% vs 0.46%, p<0.001), and less Tregs (0.88% vs 1.95%, p<0.001), compared to those who did not developed GVHD. Conclusions Patients who develop cGVHD (29%) are characterized by a pro inflammatory response with an increased production of IL-17A, IL-6, and IFN-γ, and also a major percentage of Th17 cells. Also, a decreased suppressive response was documented with reduced IL-10 and Tregs levels. The low incidence of cGVHD show that G-CSF primed bone marrow is an excellent source for allogeneic HSC transplantations, and would be useful to compare these results with other HSC sources. Disclosures No relevant conflicts of interest to declare.

scholarly journals Interferon-γ signaling synergizes with LRRK2 in neurons and microglia derived from human induced pluripotent stem cells

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Vasiliki Panagiotakopoulou ◽  
Dina Ivanyuk ◽  
Silvia De Cicco ◽  
Wadood Haq ◽  
Aleksandra Arsić ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Stem Cells ◽  
Parkinson's Disease ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Neuronal Loss ◽  
Interferon Γ ◽  
Independent Manner ◽  
Ifn Γ ◽  
Induced Pluripotent

Abstract Parkinson’s disease-associated kinase LRRK2 has been linked to IFN type II (IFN-γ) response in infections and to dopaminergic neuronal loss. However, whether and how LRRK2 synergizes with IFN-γ remains unclear. In this study, we employed dopaminergic neurons and microglia differentiated from patient-derived induced pluripotent stem cells carrying LRRK2 G2019S, the most common Parkinson’s disease-associated mutation. We show that IFN-γ enhances the LRRK2 G2019S-dependent negative regulation of AKT phosphorylation and NFAT activation, thereby increasing neuronal vulnerability to immune challenge. Mechanistically, LRRK2 G2019S suppresses NFAT translocation via calcium signaling and possibly through microtubule reorganization. In microglia, LRRK2 modulates cytokine production and the glycolytic switch in response to IFN-γ in an NFAT-independent manner. Activated LRRK2 G2019S microglia cause neurite shortening, indicating that LRRK2-driven immunological changes can be neurotoxic. We propose that synergistic LRRK2/IFN-γ activation serves as a potential link between inflammation and neurodegeneration in Parkinson’s disease.

scholarly journals Plasma Inflammatory Biomarkers Associated with Advanced Liver Fibrosis in HIV–HCV-Coinfected Individuals

2020 ◽  
Vol 17 (24) ◽  
pp. 9474
Author(s):  
Xiaochen Chen ◽  
Xing Liu ◽  
Song Duan ◽  
Renhai Tang ◽  
Sujuan Zhou ◽  
...  
Keyword(s):  
Liver Fibrosis ◽  
Plasma Concentrations ◽  
Clinical Intervention ◽  
Interferon Γ ◽  
Microbial Translocation ◽  
Cross Sectional ◽  
Monocyte Chemoattractant Protein 1 ◽  
Gm Csf ◽  
Advanced Liver Fibrosis ◽  
Ifn Γ

Background: HIV and HCV coinfection leads to accelerated liver fibrosis, in which microbial translocation and systemic inflammation might play important roles. Objective: This study aimed to provide an extensive profile of the plasma microbial translocation and inflammation biomarkers associated with advanced liver fibrosis among HIV–HCV-coinfected patients. Methods: This cross-sectional study recruited 343 HIV–HCV-coinfected patients on combination antiretroviral therapy (cART) from a rural prefecture of Yunnan province in Southwest China. The plasma concentrations of sCD14 and 27 cytokines and chemokines were assayed and compared against advanced or mild levels of liver fibrosis. Results: Of the 343 HIV–HCV-coinfected patients, 188 (54.8%) had severe or advanced liver fibrosis (FIB-4 > 3.25). The patients with advanced liver fibrosis (FIB-4 > 3.25 vs. FIB-4 ≤ 3.25) had higher plasma levels of interleukin (IL)-1β, IL-6, IL-7, IL-9, IL-12, IL-15, IL-17, granulocyte macrophage colony stimulating factor (GM-CSF), Interferon-γ (IFN-γ), tumor necrosis factor (TNF-α), IL-4, IL-10, IL-13, fibroblast growth factor 2 (FGF-basic), and Monocyte chemoattractant protein-1 (MCP-1). Multivariable logistic regression models showed that advanced liver fibrosis was associated with an increased plasma level of IL-1β, IL-6, IL-7, IL-12, IL-17, GM-CSF, IFN-γ, IL-4, IL-10, MCP-1, Eotaxin, and FGF-basic, with FGF-basic continuing to be positively and significantly associated with advanced liver fibrosis, after Bonferroni correction for multiple comparisons (adjusted odds ratio (aOR) = 1.92; 95%CI: 1.32–2.81; p = 0.001). Plasma sCD14 was also significantly associated with advanced liver fibrosis (aOR = 1.13; 95%CI: 1.01–1.30; p = 0.049). Conclusions: HIV–HCV-coinfected patients are living with a high prevalence of advanced liver fibrosis which coexists with a mixture of elevated plasma inflammation and microbial translocation biomarkers. The significant associations of advanced liver fibrosis with FGF-basic and sCD14 may reveal pathogenic mechanisms and potential clinical intervention targets for liver fibrosis in HCV–HIV coinfection.

Bone marrow-derived mesenchymal stem cells inhibit T follicular helper cell in lupus-prone mice

Lupus ◽  
2017 ◽  
Vol 27 (1) ◽  
pp. 49-59 ◽  
Author(s):  
X Yang ◽  
J Yang ◽  
X Li ◽  
W Ma ◽  
H Zou
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Flow Cytometry ◽  
T Cells ◽  
Lupus Nephritis ◽  
Bone Marrow Cells ◽  
Tfh Cells ◽  
Follicular Helper

Background The objective of this paper is to analyze the role of bone marrow-derived mesenchymal stem cells (BM-MSCs) on the differentiation of T follicular helper (Tfh) cells in lupus-prone mice. Methods Bone marrow cells were isolated from C57BL/6 (B6) mice and cultured in vitro, and surface markers were identified by flow cytometry. Naïve CD4+ T cells, splenocytes and Tfh cells were isolated from B6 mice spleens and co-cultured with BM-MSCs. The proliferation and the differentiation of CD4+ T cells and Tfh cells were analyzed by flow cytometry. Lupus-prone MRL/Mp-lpr/lpr (MRL/lpr) mice were treated via intravenous injection with expanded BM-MSCs, the differentiation of Tfh cells was detected, and the relief of lupus nephritis was analyzed. Results MSCs could be successfully induced from bone marrow cells, and cultured BM-MSCs could inhibit T cell proliferation dose-dependently. BM-MSCs could prevent Tfh cell development from naïve CD4+ T cells and splenocytes. BM-MSCs could inhibit IL-21 gene expression and cytokine production and inhibit isolated Tfh cells and STAT3 phosphorylation. In vivo study proved that BM-MSCs intravenous injection could effectively inhibit Tfh cell expansion and IL-21 production, alleviate lupus nephritis, and prolong the survival rate of lupus-prone mice. Conclusions BM-MSCs could effectively inhibit the differentiation of Tfh cells both in vitro and in vivo. BM-MSC treatment could relieve lupus nephritis, which indicates that BM-MSCs might be a promising therapeutic method for the treatment of SLE.

scholarly journals Eltrombopag maintains human hematopoietic stem and progenitor cells under inflammatory conditions mediated by IFN-γ

Blood ◽  
2019 ◽  
Vol 133 (19) ◽  
pp. 2043-2055 ◽  
Author(s):  
Luigi J. Alvarado ◽  
Heather D. Huntsman ◽  
Hai Cheng ◽  
Danielle M. Townsley ◽  
Thomas Winkler ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Binding Site ◽  
Bone Marrow Failure ◽  
Interferon Γ ◽  
General Mechanism ◽  
Clinical Activity ◽  
Critical Pathway ◽  
Hematopoietic Stem ◽  
Therapeutic Implications ◽  
Ifn Γ

Abstract The proinflammatory cytokine interferon-γ (IFN-γ) has been implicated in human hematopoietic stem and progenitor cell (HSPC) depletion in immune-mediated bone marrow failure syndromes. We show that IFN-γ specifically prevents full engagement of thrombopoietin (TPO), a primary positive regulator of HSPC survival, to its receptor (c-MPL) via steric occlusion of the low-affinity binding site, contributing to perturbation of TPO-induced signaling pathways and decreased survival of human HSPCs. Eltrombopag, a synthetic small molecule mimetic of TPO that interacts with c-MPL at a position distinct from the extracellular binding site of TPO, bypasses this inhibition, providing an explanation for its clinical activity in bone marrow failure, despite already elevated endogenous TPO levels. Thus, IFN-γ–mediated perturbation of TPO:c-MPL complex formation and the resulting inhibition of a critical pathway of growth factor cell signaling may represent a general mechanism by which IFN-γ impairs the function of human HSPCs. This understanding could have broad therapeutic implications for various disorders of chronic inflammation.

Sign in / Sign up

Export Citation Format

Share Document