Multipotent adult progenitor cells can suppress graft-versus-host disease via prostaglandin E2 synthesis and only if localized to sites of allopriming

Abstract

IntroductionThe wider application of bone marrow transplantation (BMT) has been limited, in part, by graft-versus-host disease (GVHD) complications. Human and mouse mesenchymal stem cells (MSCs) have been shown to suppress allogeneic-induced and nonspecific mitogen-induced T-cell proliferation in vitro (reviewed in detail 1,2 ). Implicated suppressive mechanisms have included interleukin (IL)-10, 3 transforming growth factor (TGF)-␤, hepatocyte growth factor, 4 indoleamine 2,3 dioxygenase (IDO), 5 nitric oxide, 6 prostaglandin (PG) E 2 , 7 increased T-regulatory cells (Tregs), 8 and activation of the PD-1-negative costimulatory pathway. 9 In vivo, there have been conflicting data regarding the potential of MSCs to suppress GVHD. [10][11][12] Multipotent adult progenitor cells (MAPCs) are nonhematopoietic stem cells that can be copurified with MSCs from BM cells. MAPCs express the pluripotent state-specific transcription factors Oct-3/4 and Rex-1, and can differentiate into cell types representative of all 3 germ layers 13,14 ; thus, MAPCs are generally believed to be a more primitive cell type than MSCs. MSCs kept for prolonged periods in culture tend to lose their differentiation capabilities and undergo senescence at approximately 20 to 40 population doublings. 15,16 In contrast to MSCs, MAPCs have an average telomere length that remained constant for up to 100 population doublings in vitro. 13 Based upon their differential potential and reduced senescence, MAPCs have been considered as a potentially desirable nonhematopoietic stem cell source for use in allogeneic BMT. In fact, a multicenter phase 1 open label clinical trial of MultiStem, based upon MAPC technology, was initiated in 2008.In this study, we sought to determine whether MAPCs might be useful for GVHD prevention. We demonstrate that murine MAPCs are potently immune suppressive in vitro and can reduce GVHD lethality in vivo when present in the spleen, a site of initial allopriming, early post-BMT. Furthermore, we identify the mechanism of action MAPCs use to elicit T-cell inhibition and reduce GVHD-induced tissue injury in vivo. Methods MiceBALB/c (H2 d ), C57BL/6 (H2 b ; termed B6), and B10.BR (H2 k ) mice were purchased from The Jackson Laboratory or the National Institutes of Health (NIH). All mice were housed in a specific pathogen-free facility in microisolator cages and used at 8 to 12 weeks of age in protocols approved by the Institutional Animal Care and Use Committee of the University of Minnesota. MAPC isolation and cultureMAPCs were isolated from B6 and BALB/c BM, as described. 13 Briefly, BM was plated in Dulbecco modified Eagle medium/MCDB containing 10 ng/mL epidermal growth factor (Sigma-Aldrich), platelet-derived growth factor-BB (R&D Systems), leukemia inhibitory factor (Chemicon International), 2% fetal calf serum (HyClone), 1ϫ selenium-insulin-transferrinethanolamine, 0.2 mg/mL linoleic acid-bovine serum albumin (BSA), 0.8 mg/mL BSA, 1ϫ chemically defined lipid concentrate, and 1ϫ ␣-mercaptoethanol (all from Sigma-Aldrich). Cells...

Clinical-Grade Multipotent Adult Progenitor Cells Durably Control Pathogenic T Cell Responses in Human Models of Transplantation and Autoimmunity

“…This result aligns with several reports highlighting that MSC-mediated suppression can occur via the IDO pathway downstream of licensing signals (e.g., IFN-g) and supports the notion that MAPCs would be most suppressive in the context of an inflammatory environment/ongoing immune response (41). Consistent with this, rodent MAPCs were shown to exhibit efficacy in the context of graft-versus-host disease only when present at sites of allopriming (36). Although implicated in MSC-mediated T cell suppression, we did not observe a role for PGE 2 in MAPC-mediated T cell suppression under any stimulation conditions.…”

“…Human MAPCs can be distinguished from human MSCs on the basis of cell surface phenotype, cell size, transcriptional profile, cytokine production, differentiation potential and replicative capacity (6, 10, 34, 35). Previous work showed that murine MAPCs exert therapeutic immunomodulatory effects in the context of graft-versus-host disease prophylaxis, in MLR systems in vitro, and that human MAPCs prevented inflammation in a murine model of stroke, supporting a role for these cells in treating human immune-mediated diseases (36)(37)(38).…”

“…Our results suggest that suppression of CD4 T cell proliferation by MAPCs imparts a cell-intrinsic long-term effect that may be bypassed when using artificial stimulation and, therefore, likely is dependent on the process of physiological Ag presentation or costimulation. Of relevance, rodent MAPCs were shown to induce CTLA-4 expression on responding T cells in MLR assays, which abrogates APCmediated costimulation but is circumvented using immobilized Abs (36). An equally plausible explanation is that long-term effects may be overcome by signal strength, either directly through enhanced signaling or indirectly by inducing de novo synthesis of endogenous IL-2.…”

Fasting and meal-stimulated residual beta cell function is positively associated with serum concentrations of proinflammatory cytokines and negatively associated with anti-inflammatory and regulatory cytokines in patients with longer term type 1 diabetes

“…We measured the proinflammatory cytokines IL-6 and TNF-α and the regulatory cytokines IL-10, TGF-β 1 , TGF-β 2 and TGF-β 3 using multiplex bead-based technology in commercially available kits (Fluorokine MAP; R&D Systems, Wiesbaden, Germany) [21][22][23]. To measure systemic concentrations of the anti-inflammatory cytokine IL-1RA, we used commercially available ELISA kits (Quantikine ELISA kits; R&D Systems).…”

Cell-Based Vascularization Strategies for Skin Tissue Engineering

“…Nevertheless, similar to MSC, MAPC also seem to have immunesuppressive capacity, which may be favorable in an allogeneic setting. 112,113 Despite their large expansion capacity, no karyotypic instabilities have been reported for human MAPC. Our recent data reveal that MAPC transplantation improves wound vascularization as well as reepithelialization in a full-thickness wound model in immunedeficient mice (unpublished results) ( Table 1).…”