Stem Cells Int. 2017; 2017: 9198328. Published online 2017 Jun 21. doi: 10.1155/2017/9198328

Intra-Articular Injection of Human Synovial Membrane-Derived Mesenchymal Stem Cells in Murine Collagen-Induced Arthritis: Assessment of Immunomodulatory Capacity In Vivo

Minglu Yan, Xin Liu, Qiujie Dang, He Huang, Fan Yang, Yang Li

Department of Rheumatology and Immunology, The Second Affiliated Hospital of Harbin Medical University, Harbin 150000, China

Correspondence should be addressed to Yang Li; liyang@hrbmu.edu.cn

 

Abstract:

The aim of this study was to evaluate the efficacy of human synovial membrane-derived MSCs (SM-MSCs) in murine collagen-induced arthritis (CIA). Male mice (age 7–9 weeks) were injected intra-articularly with SM-MSCs obtained from patients with osteoarthritis, on days 28, 32, and 38 after bovine type II collagen immunization. The efficacy of SM-MSCs in CIA was evaluated clinically and histologically. Cytokine profile analyses were performed by real-time polymerase chain reaction and multiplex analyses. Splenic helper T (Th) cell and regulatory B cell subsets were analyzed by flow cytometry. Intra-articular SM-MSC injection ameliorated the clinical and histological severity of arthritis. Decrease in tumor necrosis factor-α, interferon-γ, and interleukin- (IL-) 17A and increase in IL-10 production were observed after SM-MSC treatment. Flow cytometry showed that Th1 and Th17 cells decreased, whereas Th2, regulatory T (Treg), and PD-1+ CXCR5+ FoxP3+ follicular Treg cells increased in the spleens of SM-MSC-treated mice. Regulatory B cell analysis showed that CD21hiCD23hi transitional 2 cells, CD23lowCD21hi marginal zone cells, and CD19+ CD5+ CD1d+ IL-10+ regulatory B cells increased following SM-MSC treatment. Our results demonstrated that SM-MSCs injected in inflamed joints in CIA had a therapeutic effect and could prevent arthritis development and suppress immune responses via immunoregulatory cell expansion.

PMCID: PMC5497673

 

Supplement:

MSCs are adult multipotent cells that are present in the bone marrow (BM), adipose tissue, synovial membrane, synovial fluid, and perinatal tissues. These cells have been characterized with respect to colony-forming unit fibroblast (CFU-F), surface marker expression, and in vitro multi-differentiation potential, according to the International Society for Cellular Therapy (ISCT) criteria [1]. During the last several decades, MSCs have been largely investigated for their potent immunomodulatory and anti-inflammatory capacities, emerging as a promising therapy for autoimmune diseases such as rheumatoid arthritis (RA) . Loss of self-tolerance leads to imbalance of effector and regulatory cells, which plays a crucial role in the onset and pathogenesis of RA. Hence, the recovery of immune tolerance by expansion of regulatory cells may be a rational approach for RA treatment.

 

Synovial membrane-derived MSCs (SM-MSCs) were first identified in 2001; it was reported that the synovial membrane from the knee joints of human donors could give rise to a fibroblast-like cell population possessing great expansion potential, typical antigen expression, and multidifferentiation capability [2]. In our study, we evaluate the therapeutic effect of SM-MSCs following repeated intra-articular injection in murine collagen-induced arthritis (CIA) model.

 

In this study, human SM-MSCs were selected from among numerous cell sources because of their strong immunomodulatory properties during coculture with T lymphocytes in vitro and high proliferation capability with limited senescence. Furthermore, the “off-the-shelf” property of MSCs makes the allogeneic human-derived MSCs available for murine CIA. In addition, human synovial membranes are an accessible source of MSCs. They are routinely removed in patients with OA, during arthroscopy and knee replacement surgery, which offers the advantage of excellent supply for future clinical applications. SM-MSCs would be a promising option for the treatment of destructive diseases of the bone and cartilage.

 

Our study first showed the therapeutic effect of SM-MSCs in CIA. CIA is the most commonly studied murine model of RA and generally thought to be dependent on collagen-specific CD4+ T cells during the initial phase of autoimmune responses in the joints. Repeated administration of SM-MSCs into inflamed joints could attenuate arthritis severity with reduction in inflammatory cytokines and increase in IL-10 production in serum, suggesting that local MSC treatment could exert a systemic therapeutic effect in mice. Recent studies reported that repeated intra-articular administration of allogeneic MSCs could be a safe strategy, leading to enhanced MSC availability. It has been well documented that MSCs exert immunoregulatory effects via locally cell-cell contact or secretion of soluble modulatory mediators, where in particular MSC-derived indoleamine 2,3-dioxygenase (IDO) in human and inducible nitric oxide synthase (iNOS) in mouse [3]. Besides, the modulatory effects of MSCs on immune responses, especially by means of secreting soluble factors, are critically linked to the “license” by inflammatory signals occurred in which MSCs are applied to. We assumed that the interplay between exogenous MSCs and resident synoviocytes may trigger a series of biological processes participating in the paracrine actions in MSCs, which is most likely responsible for the therapeutic effect of MSCs in CIA.

 

Our data presented here suggest that the therapeutic effect of SM-MSCs in CIA mice was paralleled by increase in regulatory FoxP3+ T cells (Treg and Tfr) and induction of regulatory B cells (T2, MZ, and B10), both of which are essential for inhibiting dysregulated immune responses to self-antigens and are involved in self-tolerance mechanisms. Previous research documented that infusion of human gingiva-derived MSCs significantly ameliorated CIA via suppression of Th1 and Th17 responses and increase in FoxP3-expressing CD4+ T cells in the spleen [4], which is in agreement with our findings. In addition, SM-MSCs appeared to hamper the maturation and differentiation of B cells and induce the IL-10-competent regulatory B cells in our study. This was supported by the increase in immature transitional stage B cells such as CD21hiCD23hi transitional 2 (T2) cells and CD23lowCD21hi MZ cells, as well as CD5+ CD1d+ IL-10+ cells, in the spleens of SM-MSC-treated mice. Transfer of immature T2 cells from mice with arthritis at the remission stage could help recover the balance between Treg and Th1/Th17 responses in IL-10−/− hosts. Similarly, adoptive transfer of CD5+ CD1d+ IL-10+ regulatory B cells prevented CIA development in mice with suppression of Th17 cells in the spleen and draining lymph nodes. Clinical data also revealed that patients with new onset RA had lesser IL-10-competent B cells, comprising CD19+ CD5+ CD1d+ cells and CD19+ TIM1+ cells, than healthy controls and that this decrease was positively correlated with the number of CD4+ CD25+ FoxP3+ Treg cells in peripheral blood [5]. Given the above and previous data, we hypothesize that, not only FoxP3 Treg cells but also T2-, MZ-, and IL-10-expressing regulatory B cells were responsible for the suppression of inflammatory responses in mice with CIA, suggesting that the cellular interactions between regulatory B cells and Treg cells were pivotal in determining the beneficial outcome of SM-MSCs in CIA. A functional feed-forward loop between immunoregulatory T cells and B cells during SM-MSC treatment is of great interest and requires further study.

 

These findings endorse the fact that intra-articular injection of SM-MSCs could prevent arthritis development and suppress immune responses via expansion of FoxP3+ Treg cells and T2, MZ, and IL-10-competent regulatory B cells, thus recovering peripheral tolerance in mice with CIA. Our findings established the in vivo effect of SM-MSCs in CIA mice, indicating that intra-articular administration of SM-MSCs may constitute a potential approach for RA cell therapy.

 

 

Fig 1. Intra-articular injection of SM-MSCs could prevent arthritis development and suppress immune responses. SM-MSCs were able to differentiate toward osteocytes revealed by Alizarin Red S staining. We injected the prepared cells intraarticularly into the CIA mice. Repeated intra-articular injection of SM-MSCs in the right knee efficiently attenuated the arthritis symptoms, decreased the mean arthritis scores and histological damages. In addition, decrease in TNF-α, and IL-17A and increase in IL-10 production were observed after SM-MSC treatment. Th1 and Th17 cells decreased, Treg cells increased in the spleens of SM-MSC-treated mice. Regulatory B cell also increased following SM-MSC treatment. It is showed the bone regeneration and immunomodulation potential of SM-MSCs. Local MSCs therapy targeting inflamed lesion appears a profound systemic effect.

 

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