Mesenchymal Stromal Cells from the Epidermis and Dermis of Psoriasis Patients: Morphology, Immunophenotype, Differentiation Patterns, and Regulation of T Cell Proliferation

Psoriasis is a skin disease characterized by hyperproliferation of keratinocytes and chronic inflammation. Mesenchymal stem/stromal cells (MSCs) exhibit an immunoregulatory function that can be altered in the skin of these patients. However, to date, the presence and functional capacity of MSCs in the dermis and epidermis of patients with psoriasis have not been fully established. In the present study, we evaluated the presence of MSCs in the skin of patients by obtaining adherent cells from the dermis and epidermis of lesional and nonlesional areas and characterizing them in a comparative manner with corresponding cells obtained from the dermis (HD-MSCs) and epidermis (HE-MSCs) of healthy donors. We determined whether the adherent cells had immunophenotypic profiles and differentiation potentials that were characteristic of MSCs. In addition, we analyzed their immunosuppression function by evaluating their capacity to decrease T cell proliferation. Our results indicate the presence of MSCs in the dermis and epidermis of healthy donors and patients with psoriasis; adherent cells from all skin sources exhibited MSC characteristics, such as expression of CD73, CD90, and CD105 markers and a lack of hematopoietic and endothelial marker expression. However, the cell populations obtained showed differences in differentiation potential toward adipogenic, osteogenic, and chondrogenic lineages. In addition, we observed a low MSC obtention frequency in nonlesional epidermal samples (NLE-MSCs), which also showed alterations in morphology and proliferation rate. Interestingly, MSCs from both the nonlesional dermis (NLD-MSCs) and lesional dermis (LD-MSCs) showed higher HLA class I antigen (HLA-I) expression than HD-MSCs. Moreover, NLD-MSCs showed a low T cell proliferation suppression capacity. In summary, this study demonstrates the presence of MSCs in the epidermis and dermis of patients with psoriasis and suggests that such cells may favor the inflammatory process and thus psoriatic lesion development through high HLA-I expression and low immunosuppression capacity.

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Fas-Independent Apoptosis of Activated T Cells Induced by Antibodies to the HLA Class I α1 Domain

Blood ◽

10.1182/blood.v90.9.3629 ◽

1997 ◽

Vol 90 (9) ◽

pp. 3629-3639 ◽

Cited By ~ 32

Author(s):

Laurent Genestier ◽

Romain Paillot ◽

Nathalie Bonnefoy-Berard ◽

Geneviéve Meffre ◽

Monique Flacher ◽

...

Keyword(s):

Cell Proliferation ◽

T Cells ◽

T Cell ◽

T Lymphocytes ◽

Heavy Chain ◽

Hla Class I ◽

Class I ◽

T Cell Proliferation ◽

Activated T Cells ◽

Hla Class I Molecules

Abstract In addition to their major function in antigen presentation and natural killer cell activity regulation, HLA class I molecules may modulate T-cell activation and proliferation. Monoclonal antibodies (MoAbs) that recognize distinct epitopes of HLA class I molecules were reported to interfere with T-cell proliferation. We show here that two MoAbs (mouse MoAb90 and rat YTH862) that bind to an epitope of the α1 domain of HLA class I heavy chain induce apoptotic cell death of activated, but not resting, peripheral T lymphocytes. Other reference anti-HLA class I antibodies specific for distinct epitopes of the α1 (B9.12.1), α2 (W6/32), or α3 (TP25.99) domains of the heavy chain decreased T-cell proliferation but had little or no apoptotic effect. Apoptosis shown by DNA fragmentation, phosphatidylserine externalization, and decrease of mitochondrial transmembrane potential was observed whatever the type of T-cell activator. Apoptosis did not result from Fas/Fas-L interaction and distinct though partly overlapping populations of activated T cells were susceptible to Fas– and HLA class I–mediated apoptosis, respectively. Induction of apoptosis did not require HLA class I cross-linking inasmuch as it could be observed with monovalent Fab′ fragments. The data indicate that MoAb90 and YTH862 directed against the α1 domain of HLA class I trigger apoptosis of activated T lymphocytes by a pathway which does not involve Fas-ligand.

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Delta-Like-1 Changes the Immunomodulatory Property of OP9 Cells

Stem Cells International ◽

10.1155/2016/1628352 ◽

2016 ◽

Vol 2016 ◽

pp. 1-11

Author(s):

Lei Zhang ◽

Rui-Jie Dang ◽

Yan-Mei Yang ◽

Dian-Chao Cui ◽

Ping Li ◽

...

Keyword(s):

Cell Proliferation ◽

T Cell ◽

B Cell ◽

Stromal Cells ◽

B Lymphocyte ◽

B Cell Development ◽

T Cell Proliferation ◽

No Production ◽

Immunomodulatory Property

As stromal cells and recently confirmed mesenchymal stem cells, OP9 cells support hematopoiesis stem cell (HSC) differentiation into the B lymphocyte lineage, yet Delta-like-1 (DL1) overexpressing OP9 (OP9DL1) cells promote the development of early T lymphocytes from HSC. However, the immunomodulatory capacity of OP9 or OP9DL1 on mature B and T cell proliferation has not been elucidated. Here, we show that OP9 and OP9DL1 have similar proliferation capacities and immunophenotypes except DL1 expression. Compared with OP9, OP9DL1 displayed more osteogenesis and less adipogenesis when cultured in the respective induction media. Both OP9 and OP9DL1 inhibited mature B and T cell proliferation. Furthermore, OP9 showed stronger inhibition on B cell proliferation and OP9DL1 exhibited stronger inhibition on T cell proliferation. With stimulation, both OP9 and OP9DL1 showed increased nitrate oxide (NO) production. The NO levels of OP9 were higher than that of OP9DL1 when stimulated with TNFα/IFNγor LPS/IL4. Taken together, our study reveals a previously unrecognized role of OP9 and OP9DL1 in mature B and T cell proliferation. DL1 overexpression alone changed the properties of OP9 cells in addition to their role in early B cell development.

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A Population of Suppressive Monocytes Inhibiting T Cell Proliferation and Dendritic Cell Differentiation in Relapsed Non-Hodgkin’s Lymphoma

Blood ◽

10.1182/blood.v112.11.808.808 ◽

2008 ◽

Vol 112 (11) ◽

pp. 808-808

Author(s):

Yi Lin ◽

Peggy A Bulur ◽

Michael P. Gustafson ◽

Thomas E. Witzig ◽

Allan B. Dietz

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Peripheral Blood ◽

Hodgkin’S Lymphoma ◽

Hodgkin's Lymphoma ◽

T Cell Proliferation ◽

Hla Dr ◽

Healthy Donors ◽

Non Hodgkin's Lymphoma ◽

Kinase Phosphorylation

Abstract Despite advances in treatments for patients with non-Hodgkin’s lymphoma (NHL), the relapse rates remain high and 40% of diffuse large B-cell NHL (DLBCL) patients die of disease. New therapies to augment the host anti-tumor immune response are needed. Reports of graft-versus-lymphoma responses in patients who have received allogeneic hematopoietic cell transplant indicate a role for cellular immunotherapy. However, these patients have variable levels of immunodeficiency which may impact the efficacy of cellular therapy. To study this we first evaluated the cellular immune status of patients with relapsed NHL. Proliferation of peripheral blood mononuclear cells (PBMNC) stimulated with anti-CD3/CD28 beads was reduced by more than 3.5 folds for patients with DLBCL (n = 3) compared to that of age-matched healthy donors (n = 5; p = 0.02). Removal of monocytes from PBMNC by use of anti-CD14 immunomagnetic beads restored proliferation to that of healthy donors. Further, monocytes from these patients were deficient in stimulating allogeneic T cell proliferation by 3 folds compared to monocytes from healthy donors (n = 3 NHL; n = 8 normal; p < 0.01). Peripheral blood from 12 NHL patients (9 DLBCL; 1 grade 3 follicular lymphoma; 2 composite) and 12 age-matched healthy donors were characterized by flow cytometry to determine the phenotype of these suppressive monocytes. There was no difference in the % monocytes in the blood between NHL patients and healthy donors; however, NHL patients had elevated % monocytes with a suppressive phenotype (CD14+HLA-DRneg) compared to normals (NHL 38.9 ± 4.93%; normal 8.3 ± 2.15; p < 0.0001). This phenotype is distinct from other myeloid suppressors (Lin-CD33+HLA-DR-) or non-classical monocytes (CD16+), neither of which was different in numbers between NHL and normal donors. This suggests that the CD14+HLA-DRneg monocytes are responsible for the observed T cell suppression. To further characterize the function of these cells, we cultured purified CD14+ monocytes from NHL and compared their differentiation capacity with those from normal donors. The percentage of CD14+HLA-DRneg monocytes in initial ex vivo culture was inversely correlated with the percentage of pure, mature dendritic cells (mDC) generated with TNF-a and PGE2 as maturation factors (CD80+CD83+; n = 9; p = 0.015). As CpG oligonucleotides are also capable of immune stimulation and have some anti-tumor activity in clinical trials, we investigated the effect of CpG on mDC differentiation in NHL. In healthy donors, maturation of monocytes with CpG yielded highly pure mDC (90.7 ± 2.15%, n = 3). However, preliminary results of mDC yield from monocytes of NHL patients matured with CpG was only 22.6 ± 11.2% (n = 2). This data suggests alternative signaling pathways of these suppressive monocytes. Preliminary analysis of a proteome array for 46 kinase phosphorylation sites from 37 proteins in 2 NHL and 3 healthy donors suggest changes in phosphorylation of 17 protein kinases for CD14+HLA-DRneg compared to CD14+HLA-DR+ cells. Functional correlation of these protein kinase phosphorylation changes is needed to definitively target the pathway characteristic of CD14+HLA-DRneg monocytes. Finally, we have identified a serum-free culture method that can consistently generate highly pure mDC from monocytes of NHL patients (93.0 ± 3.6%, n = 9) and is readily adaptable to good manufacturing practice for clinical use in immunotherapy. Taken together we have described for the first time a population of CD14+HLA-DRneg monocytes that is a significant source of immunosuppression in NHL patients and are beginning to target methods of overcoming this suppression.

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Inhibition of T-Cell Proliferation by Murine Multipotent Mesenchymal Stromal Cells is Mediated by CD39 Expression and Adenosine Generation

Cell Transplantation ◽

10.3727/096368910x546553 ◽

2011 ◽

Vol 20 (8) ◽

pp. 1221-1230 ◽

Cited By ~ 52

Author(s):

Christine Sattler ◽

Manuela Steinsdoerfer ◽

Monika Offers ◽

Elke Fischer ◽

Rudolf Schierl ◽

...

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Multipotent Mesenchymal Stromal Cells ◽

T Cell Proliferation

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Placenta Derived Adherent Cells Modulate the Allogenic Mixed Lymphocyte Reaction Partially via an Indoleamine 2,3-Dioxygenase Mediated Mechanism.

Blood ◽

10.1182/blood.v110.11.4112.4112 ◽

2007 ◽

Vol 110 (11) ◽

pp. 4112-4112

Author(s):

Bitao Liang ◽

Casper Paludan ◽

Matthew Downey ◽

Craig Lewis ◽

Ryhor Harbacheuski ◽

...

Keyword(s):

Cell Proliferation ◽

T Cells ◽

T Cell ◽

Conditioned Media ◽

T Cell Proliferation ◽

Adherent Cells ◽

Soluble Factors ◽

Naive T Cells ◽

Naïve T Cells ◽

Bottom Chamber

Abstract Placenta Derived Adherent Cells (PDAC) are multipotent progenitor cells derived from human placental tissues. Previously we have reported that PDACs could suppress T-cell proliferation when added to in-vitro mixed lymphocyte reactions (PDAC-MLR) (Paludan C. et al, Blood. (ASH Annual Meeting Abstracts) 2006 108: Abstract 1737). Here we present aspects of the mechanism of this PDAC suppression property. We have found that PDACs modify cytokine production in the PDAC-MLR reaction in comparison to the MLR; TNF-α and IFN-γ are reduced 75% and 30% while TGF-β production is significantly increased. We have used a transwell assay system to investigate the cell-contact-dependency of the effects of PDACs on T-cell proliferation. The assay system comprised combinations of the MLR in the top chamber together with the PDAC-MLR, PDACs plus naive T cells or PDACs alone in the bottom chamber. Maximum inhibition of T cell proliferation of the MLR in the top insert could be achieved by placing the PDAC-MLR co-culture in the bottom chamber. Minimum suppression was obtained when placing PDACs plus naive T cells or PDACs alone in the bottom chamber. PDAC-MLR conditioned media could partially suppress the MLR reaction. Addition of L-tryptophan into the MLR with PDAC conditioned media completely abolished PDAC-induced suppression of T cell proliferation. Likewise, the addition of the 1-methyl tryptophan to the PDAC-MLR reaction could abolish the PDAC-induced suppression. These results suggested that the suppression of the MLR by PDACs was possibly due to the depletion of the essential amino acid L-tryptophan which could be due to up-regulation of indoleamine 2,3-dioxygenase (IDO). Quantitative RT-PCR analysis of IDO gene expression revealed that IDO was up-regulated by about 4000-fold when PDACs were co-cultured with activated T cells, but not when co-cultured with naive T cells. Experiments are ongoing to confirm the causative role of IDO, and other factors, in PDAC-suppression of T-cell proliferation. In summary, we believe that soluble factors including the production of pro-inflammatory cytokines may contribute to PDAC suppression of the MLR, that induction of soluble factors from PDACs is significantly augmented by T-cell activation and that IDO expression by PDACs during the PDAC-MLR reaction plays a significant and direct role in suppression of T cell proliferation by PDACs.

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Membrane-Associated Proteinase 3 on Granulocytes and Myeloid Leukemia Mediates Reversible Inhibition of T Cell Proliferation

Blood ◽

10.1182/blood.v118.21.1916.1916 ◽

2011 ◽

Vol 118 (21) ◽

pp. 1916-1916

Author(s):

Tian-Hui Yang ◽

Karen Clise-Dwyer ◽

Gheath Alatrash ◽

Kathryn Ruisaard ◽

Shoudan Liang ◽

...

Keyword(s):

Cell Proliferation ◽

T Cells ◽

T Cell ◽

Cd4 T Cells ◽

Myeloid Leukemia ◽

Inhibitory Effect ◽

T Cell Proliferation ◽

Proteinase 3 ◽

Healthy Donors ◽

Membrane Bound

Abstract Abstract 1916 Proteinase 3 (P3), a serine protease constitutively expressed in primary granules and on the membrane of some resting granulocytes, is the target of T cell-mediated autoimmunity in Wegener's granulomatosis (WG) and of anti-leukemia immunity mediated by PR1 (VLQELNVTV)-specific cytotoxic T lymphocytes (PR1-CTL). We have shown that soluble P3 is increased by 5-fold in sera from acute myeloid leukemia (AML) patients compared to healthy controls, and soluble P3 mediates enzyme-independent inhibition of T-cell proliferation. Moreover, tumor-associated neutrophils (TANs) are associated with a poor prognosis in a number of cancers including renal cell carcinoma and lung cancer, and P3 is also over-expressed in a variety of AML and chronic myeloid leukemia (CML). Therefore, we hypothesized that membrane-bound P3 (mP3) may similarly regulate adaptive immunity by suppressing T-cell proliferation. To study this, T cells from healthy donors were labeled with the membrane dye CFSE and stimulated with anti-CD3 and anti-CD28 in the presence or absence of mP3-expressing PMNs for five days. The percentage of proliferating cells was determined by flow cytometry. Proliferation of autologous CD8+ and CD4+ T cells was significantly inhibited by 75% and 72%, respectively, when PMNs were co-incubated with lymphocytes at a ratio of 3:1, and by > 90% at 5:1. This cell contact-dependent inhibitory effect was limited to PMN since PBMCs added to lymphocytes in place of PMNs at 5:1 had no effect on T cell proliferation. To determine whether the inhibitory effect was specifically mediated by mP3, we FAC-sorted CD177+PMNs and CD177−PMNs to obtain highly purified (>98%) mP3+ and mP3− PMNs, respectively, because CD177 and mP3 are co-expressed on same subset of resting PMNs. At a ratio of 3:1 (CD177+PMNs or CD177−PMNs to lymphocytes), mP3+PMNs mediated > 75% growth inhibition of both CD8+ and CD4+ T cells compared to < 55% inhibition by mP3−PMNs (p<0.05). Furthermore, the inhibitory effect of mP3+PMNs on T cell proliferation was blocked (< 10% inhibition of proliferation) by anti-P3 but not by isotype control mAb. The inhibitory effect of mP3 was enzyme-independent because Elafin or α1-anti-trypsin did not affect inhibition by mP3+PMNs. In addition, mP3-mediated inhibition was fully reversible as T cells proliferated normally with anti-CD3/anti-CD28 stimulation after PMNs were removed from co-culture. Similarly, mP3+AML blasts inhibited autologous CD8+ and CD4+ T cell proliferation by 50% and 30%, respectively, at a 2:1 ratio of AML blasts: lymphocytes. Interestingly, bone marrow myeloid derived suppressor cells (MDSC) from leukemia patients express significantly higher mP3 (79.4±5.23% (mean±SEM, n=7)), compared to 22.4±11.55% mP3 on MDSC from healthy donors (p= 0.0007, n=3). Taken together, these data support an important new function of membrane-bound P3 on innate immune cells and leukemia in controlling adaptive T cell immunity. These findings suggest a novel mechanism whereby neutrophils could promote tumor growth in vivo through contact-mediated suppression of tumor-infiltrating lymphocytes by mP3 or by soluble P3 secreted by activated TANs in cancer and myeloid leukemia. Thus, targeting P3 with anti-P3 antibodies may be explored as a novel therapeutic strategy for leukemia and other cancers. Disclosures: No relevant conflicts of interest to declare.

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Human multipotent mesenchymal stromal cells cytokine priming promotes RAB27B-regulated secretion of small extracellular vesicles with immunomodulatory cargo

Stem Cell Research & Therapy ◽

10.1186/s13287-020-02050-6 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Anastasia Cheng ◽

Dongsic Choi ◽

Maximilien Lora ◽

Dominique Shum-Tim ◽

Janusz Rak ◽

...

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Extracellular Vesicles ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Multipotent Mesenchymal Stromal Cells ◽

Clinical Applications ◽

T Cell Proliferation ◽

Dependent Manner ◽

Soluble Factors

Abstract Background The paracrine effects of multipotent mesenchymal stromal cells (MSCs) are mediated by their secretome composed by soluble factors (i.e., cytokines, growth factors, hormones) and extracellular vesicles (EVs). EVs promote intercellular communication, and the EV cargoes [e.g., proteins, soluble factors, microRNAs (miRNAs), messenger RNA (mRNA), DNA] reflect the molecular and functional characteristics of their parental cells. MSC-derived EVs (MSC-EVs) are currently evaluated as subcellular therapeutics. A key function of the MSC secretome is its ability to promote immune tolerance (i.e., immunopotency), a property that is enhanced by priming approaches (e.g., cytokines, hypoxia, chemicals) and inversely correlates with the age of the MSC donors. We evaluated mechanisms underlying MSC vesiculation and the effects of inflammation and aging on this process. Methods We evaluated the effects of interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α) on human adipose-derived MSC: (a) vesiculation (custom RT2 Profiler PCR Array), (b) EV profiles (Nanoparticle Tracking Analysis and Nanoparticle Flow Cytometry), (c) EV cargo (proteomic analysis and Western blot analysis), and (d) immunopotency (standard MSC:CD4 T cell proliferation inhibition assay). We confirmed the role of RAB27B on MSC vesiculation (RAB27B siRNA) and assessed its differential contribution to vesiculation in adult and pediatric MSCs (qPCR). Results Cytokine priming upregulated RAB27B in adipose-derived MSCs increasing their secretion of exosome-like small EVs (sEVs; < 200 nm) containing two key mediators of immunopotency: A20 and TSG-6. These EVs inhibited T cell proliferation in a dose-dependent manner. RAB27B siRNA inhibited MSC vesiculation. Adipose-derived MSCs isolated from pediatric donors exhibited higher RAB27B expression and secreted more sEVs than adult MSCs. Conclusions Cytokine priming is a useful strategy to harvest anti-inflammatory MSC-sEVs for clinical applications. Of relevance, donor age should be considered in the selection of MSC-sEVs for clinical applications.

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Proinflammatory stimuli induce galectin-9 in human mesenchymal stromal cells to suppress T-cell proliferation

European Journal of Immunology ◽

10.1002/eji.201343335 ◽

2013 ◽

Vol 43 (10) ◽

pp. 2741-2749 ◽

Cited By ~ 57

Author(s):

Friederike Gieseke ◽

Anne Kruchen ◽

Nikolay Tzaribachev ◽

Frank Bentzien ◽

Massimo Dominici ◽

...

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

T Cell Proliferation ◽

Human Mesenchymal Stromal Cells

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The suppressive effect of mesenchymal stromal cells on T cell proliferation is conserved in old age

Transplant Immunology ◽

10.1016/j.trim.2011.06.007 ◽

2011 ◽

Vol 25 (2-3) ◽

pp. 167-172 ◽

Cited By ~ 16

Author(s):

Katja Landgraf ◽

Regina Brunauer ◽

Günter Lepperdinger ◽

Beatrix Grubeck-Loebenstein

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Old Age ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Suppressive Effect ◽

T Cell Proliferation

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Mesenchymal Stromal Cells from Osteoarthritic Synovium Are a Distinct Population Compared to Their Bone-Marrow Counterparts regarding Surface Marker Distribution and Immunomodulation of Allogeneic CD4+ T-Cell Cultures

Stem Cells International ◽

10.1155/2016/6579463 ◽

2016 ◽

Vol 2016 ◽

pp. 1-17 ◽

Cited By ~ 2

Author(s):

Sebastien Hagmann ◽

Claudia Rimmele ◽

Florin Bucur ◽

Thomas Dreher ◽

Felix Zeifang ◽

...

Keyword(s):

Bone Marrow ◽

Cell Proliferation ◽

T Cells ◽

T Cell ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Cd4 T Cells ◽

Surface Marker ◽

Cd4 T Cell ◽

T Cell Proliferation

Introduction. The participation of an inflammatory joint milieu has been described in osteoarthritis (OA) pathogenesis. Mesenchymal stromal cells (MSCs) play an important role in modulating inflammatory processes. Based on previous studies in an allogeneic T-cell coculture model, we aimed at further determining the role of synovial MSCs in OA pathogenesis.Methods. Bone-marrow (BM) and synovial membrane (SM) MSCs from hip joints of late stage OA patients and CD4+ T-cells from healthy donors were analysed regarding surface marker expression before and after coculture. Proliferation upon CD3/CD28 stimulation and cytokine analyses were compared between MSCs.Results. SM-MSCs differed from BM-MSCs in several surface markers and their osteogenic differentiation potential. Cocultures of both MSCs with CD4+ T-cells resulted in recruitment of CD45RA+ FoxP3+ regulatory T-cells. Upon stimulation, only SM-MSCs suppressed CD4+ T-cell proliferation, while both SM-MSCs and BM-MSCs modified cytokine profiles through suppressing IL-2 and TNF-αas well as increasing IL-6 secretion.Conclusions. Synovial MSCs from OA joints are a unique fraction that can be distinguished from their bone-marrow derived counterparts. Their unique ability to suppress CD3/CD28 induced CD4+ T-cell proliferation makes them a potential target for future therapeutic approaches.

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