Bone Marrow Mesenchymal Stromal Cells from Patients with Acute and Chronic Graft-versus-Host Disease Deploy Normal Phenotype, Differentiation Plasticity, and Immune-Suppressive Activity

Abstract

The success of allogeneic hematopoietic stem cell transplantation (allo-HSCT) is often limited by the development of acute and/or chronic graft-versus-host disease (GVHD). The lack of effective therapies to treat steroid-refractory GVHD patients has bolstered clinical evaluation of mesenchymal stromal cell (MSC) therapy for GVHD. Currently, testing of MSCs for the treatment of GVHD has exclusively used allogeneic MSCs despite emerging evidence that MSCs lose their immunoprivileged status in vivo. We hypothesized that autologous MSCs could be a viable alternative MSC source for treating active GVHD. MSCs were isolated and successfully expanded from the bone marrow of 12 volunteers (ages 2 to 55 years) who had allo-HSCT transplants and subsequently developed GVHD. MSCs from subjects with GVHD demonstrated an initial lag in growth compared with healthy control subjects; however, this lag disappeared with continued ex vivo expansion. Immunophenotype and mesodermal differentiation capacity of MSCs from GVHD patients were indistinguishable from that of healthy control MSCs. In vitro immunomodulatory functional analyses also demonstrated that GVHD MSCs were equivalent to healthy control MSCs with regards to dose dependently suppressing T cell proliferation and up-regulating indoleamine 2,3-dioxygenase expression when primed with IFN-γ. Single tandem repeat chimerism analyses further demonstrated that MSCs expanded from GVHD patients were exclusively recipient derived. Based on these data, we conclude that recipient-derived MSCs from patients with GVHD are analogous to MSCs from healthy volunteers and represent a viable option for clinical testing as an immunomodulatory option for symptomatic GVHD.

Immunomodulatory effects of OX40Ig gene-modified adipose tissue-derived mesenchymal stem cells on rat kidney transplantation

“…However, the cytogenetic stability of MSCs following viral transduction needs to be established to the allay safety issues of malignant transformation. The nucleofection technology is a safe non-viral electroporation-based transfection system (24,25). In this study, we modified ADSCs with the plasmid pcDNA3.1 that can expressed the OX40Ig fusion protein in eukaryotic expressiion systems by nucleofection.…”

Potency Analysis of Mesenchymal Stromal Cells Using a Combinatorial Assay Matrix Approach

“…Patients enrolled in this study had active GvHD or inflammatory bowel disease at the time of bone marrow harvest, as described previously (Copland et al, 2015; Dhere et al, 2016). Bone marrow aspirates were subjected to a Ficoll density gradient to obtain mononuclear cells, as described previously (Chinnadurai et al, 2014).…”

Morphological features of IFN-γ–stimulated mesenchymal stromal cells predict overall immunosuppressive capacity

“…Another significant challenge facing development of MSC-based therapies is the lack of well-defined, robust assays for assessing their immunosuppressive function. Current assays for assessing MSC immunosuppressive capacity exist; however, they often rely on quantification of only a few markers in a single culture condition (8,9,20). Based on these limitations, we developed a robust, quantitative method of assessing the immunosuppressive capacity of human bone marrow-derived MSCs and further demonstrated that morphological features of IFN-γ-stimulated MSCs can be used to predict their overall immunosuppressive capacity.…”

“…A major challenge in the development of consistently effective MSC-based immunosuppressive therapies is that MSC lines derived from different donors and manufacturing processes (i.e., cell expansion) can possess markedly dissimilar immunosuppressive function (3,5,6). Although methods exist to assess MSC immunosuppression in vitro, they are often based on only a few measured outcomes, assay culture conditions, and donor MSC samples (5,(7)(8)(9). To improve upon these methods, we developed an experimental and analytical approach to quantify MSC-mediated immune suppression using principal-component analysis (PCA) to integrate multiple measurements of T-cell activation assessed at a range of MSC densities.…”