人间充质的效果干从狼疮易感MRL隔绝在异种T细胞和B细胞.Fas^LPR老鼠

抽象

系统性红斑狼疮（SLE）是一种自身免疫性疾病，其特征是T和B细胞的过度活化。人间充质干细胞（hMSCs），使用狼疮易感MRL改善SLE的进展的临床前研究.Fas^LPR老鼠。然而，无论的hMSCs抑制异种小鼠T和B细胞的功能尚不清楚。为了解决这个问题，我们研究了in vitro对T的hMSC细胞和B细胞的作用从分离MRL.Fas^LPR老鼠。幼稚的hMSCs抑制T细胞的功能，但不B细胞。hMSCs的预充IFN-γγ（一世）inhibited the proliferation of and IgM production by B cells, (ii) attracted B cells for cell–cell interactions in a CXCL10-dependent manner, and (iii) inhibited B cells by producing indoleamine 2,3-dioxygenase. In summary, our data demonstrate that hMSCs exert therapeutic activity in mice in three steps: first, naïve hMSCs inhibit the functions of T cells, hMSCs are then activated by IFN-γ，最后，它们抑制B细胞。

1。Introduction

Mesenchymal stem cells (MSCs) are multipotent progenitor cells and have broad immunoregulatory properties on T cells (including Tregs), B cells, and dendritic cells [1]。的MSC是公知的通过产生可溶性因子，如肿瘤生长因子抑制T细胞的功能（TGF-β）β，前列腺素E₂（PGE₂），和吲哚胺2,3-双加氧酶（IDO），并通过表达FasL和PD-L1为接触依赖性抑制，以诱导T细胞凋亡[2，3]。However, the effect of MSCs on B cells is controversial: MSCs inhibited proliferation, antibody production, and migration of B cells in some studies [4-6]，但都没有效果，甚至在其他国家增加了这些功能[7-11]。It was also reported that MSCs indirectly inhibit B cell proliferation by suppressing helper T cell functions [7，12]。Generally, the inhibitory mechanisms of MSCs have been studiedin vitroby using species-matched autologous and allogeneic target immune cells, but not species-mismatched xenogeneic immune cells.

系统性红斑狼疮（SLE）的特征在于，产生自身抗体，以普遍存在的自身抗原[的13]。In preclinical studies, the transfer of human MSCs (hMSCs) to lupus-prone MRL/MpJ-FAS^LPR（称为MRL。FAS^LPR下文）小鼠增加了它们的生存和降低抗dsDNA抗体水平和肾炎[14]。However, it is still unclear whether hMSCs inhibit mouse T and B cells in this xenogeneic animal model. Here, we addressed this issue and investigated the interaction between hMSCs and mouse B cells in detail, since B cells play a critical role in SLE pathogenesis [13]。We found that naïve hMSCs inhibited mouse T cells only, whereas priming of hMSCs with IFN-γrendered them capable of inhibiting mouse B cells in a CXCL10- and IDO-dependent manner.

2。Materials and Methods

2。1。Mesenchymal Stem Cells

Human bone marrow-derived MSCs were obtained from Corestem Inc. (Seoul, Korea) [1五]。简言之，骨髓从健康供体的后髂嵴吸出，并通过密度梯度离心收集单核细胞。These cells were cultured in CSMB-A06 medium (Corestem Inc.) containing 10% fetal bovine serum (BD Biosciences, Franklin Lakes, NJ, USA), 2.5 mM大号-glutamine, and penicillin/streptomycin (WelGene, Gyeonggi, Korea) in a 5% CO₂incubator at 37°C for 3–5 passages. After washing out nonadherent cells, the adherent cells retained the canonical phenotype of MSCs (CD29⁺CD44⁺CD73⁺CD105⁺CD90⁺CD34⁻CD45⁻HLA-DR⁻）and were used in the experiments [16]。All human MSC studies were approved by the Institutional Review Board of Hanyang University Hospital and were carried out in accordance with the approved guidelines; all participants provided written informed consents.

2.2。狼疮易感MRL。FAS^LPR老鼠

女性MRL.Fas^LPRmice were purchased from the Jackson Laboratory (Bar Harbor, MA, USA). Mice were housed in specific pathogen-free conditions at 21–24°C and 40–60% relative humidity under a 12 h light/dark cycle. Female MRL.Fas^LPR小鼠用PBS（载体静脉内注射， ）or hMSCs/mouse ( ）once at the age of 12 weeks [14]。Survival rate and body weight were examined every week. Serum was collected every 3 weeks and stored at −70°C until use. The levels of anti-dsDNA IgG and total IgG in serum were measured by using ELISA kits purchased from Alpha Diagnostic International (San Antonio, TX, USA) and eBiosciences (San Diego, CA, USA), respectively, according to the manufacturers’ instructions. All animal studies were approved by the Chungbuk National University Animal Experimentation Ethics Committee and were carried out in accordance with the approved guidelines.

2.3。RNA干扰

siRNAs targeting mouse chemokines were purchased from Bioneer (Daejeon, Korea). The following sequences were used: CCL2 (GenBank accession number NM 002982.3), 5 -CUC CGA AGA CUU GAA CAC UdTdT-3 ，五 -GAA ACA UCC AAU UCU CAA AdTdT-3 ，五 -GCU CGC GAG CUA UAG AAG AdTdT-3 ;CXCL10 (GenBank accession number NM 001565.3), 5 -GGU CAC CAA AUC AGC UGC UdTdT-3 ，五 -GAG AUC AUU GCU ACA AUG AdTdT-3 ，五 -CAU GAA UCA AAC UGC CAU UdTdT-3 ;CXCL12 (GenBank accession number NM 199168.4), 5 -GAU UCU UCG AAA GCC AUG UdTdT-3 ，五 -CCA GAG CCA ACG UCA AGC AdTdT-3 ，五 -CAA CAG ACA AGU GUG CAU UdTdT-3 。下面负对照siRNA使用：5 -CCU ACG CCA CCA AUU UCG UdTdT-3 。MSCs were transfected with 100 nM siRNAs using Lipofectamine RNAiMAX reagent (Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer’s protocol. Cells were incubated at 37°C in a 5% CO₂incubator for 48 h [17]。

2。4。Transwell Assay and Time-Lapse Imaging

在转孔试验中，B细胞（100 μ升）加入到Transwell小板的上层孔用5 μ米插入物（Corning公司，Tewksbury的，MA，USA）。趋化因子或MSC的各种浓度加入到在600的低孔 μ升的完全RPMI 1640培养基中。The number of B cells that had migrated to the lower well over 1.5 h was counted using a flow cytometer [14]。

对于时间推移成像，的MSC（70 μl of cells/ml) were seeded into the left chamber and B cells (70 μl of 细胞/ ml）到培养嵌件的右腔室μ-Dish^3五mm培养皿（ibidi有限公司，马丁斯里德，德国）。时间推移成像用在保持在37℃和5％CO的环境室配备有10倍的放大倍数的物镜（数值孔径0.5）一个BioStation的IM-Q显微镜进行₂（尼康公司，梅尔维尔，NY，USA）。Dishes were preincubated for 3 h in the chamber, and then, inserts were carefully removed. Images were acquired every 2 min for 12 h [18]。

2.5。B和T细胞功能

B cells were purified from spleen cells of MRL.Fas^LPRmice by a negative depletion method using a B cell isolation kit (Miltenyi Biotec, Auburn, CA, USA). T cells were purified using a T cell isolation kit (Miltenyi Biotec). Purity of both cell types was 。To measure B cell proliferation, purified B cells ( 细胞/孔）和间充质干（ 细胞/孔）在96孔板中混合。在一些实验中，间充质干在下部孔transwell板（Corning）中，以避免细胞 - 细胞接触。的上孔和B细胞进行培养14]。B cells were activated with lipopolysaccharide (LPS, 1 μg/ml) on day 0 [19]。³H-Thymidine (113 Ci/nmol; NEN, Boston, MA, USA) was added to the culture at a concentration of 1 μCi/well 54 h after LPS stimulation, and B cells were harvested using an automated cell harvester (Inotech, Dottikon, Switzerland) 72 h after LPS stimulation. The amount of³掺入细胞内H-胸苷使用Wallac Microbeta闪烁计数器（Wallac，图尔库，芬兰）测量。The levels of IgM accumulated in culture medium for 72 h were determined by using an ELISA kit (Thermo Fisher Scientific). To measure T cell proliferation, concanavalin A (Con A, 1 μg/ml) was used to specifically activate T cells [19]。The levels of T cell-derived IFN-γwere determined by using an ELISA kit (Bio-Techne, Minneapolis, MN, USA).

2。6。Western Blotting, RT-PCR, Quantitative PCR (qPCR), and ELISA

对于蛋白质印迹，如先前所述制备细胞裂解物[20]。除去洗涤剂溶性物质，并将等量的蛋白通过10％SDS-PAGE分级分离并转移至纯硝酸纤维素膜上。Membranes were blocked with TBS/Tween 20 containing 5% bovine serum albumin for 1 h and then incubated with an appropriate dilution of primary antibody in the same buffer for 2 h. Blots were incubated with biotinylated secondary antibody for 1 h and then with HRP-conjugated streptavidin for 1 h. Signals were detected by enhanced chemiluminescence (Amersham Pharmacia Biotech, Piscataway, NJ, USA). Anti-mouse or anti-human antibodies against STAT1, phospho-STAT1, IDO, and GAPDH were purchased from Cell Signaling Technology (Danvers, MA, USA).

用于RT-PCR总RNA，使用TRIZOL试剂（赛默飞世尔科技）的MSC分离。RNA was quantified using a spectrophotometer and stored at –80°C at a concentration of 1 mg/ml. cDNA was synthesized from 3 μg total RNA using an RT kit (Bioneer). PCR was used to examine the expression levels of the following mRNAs: COX-2, TGF-β，IDO, CCL2, CCL3, CCL5, CXCL10, and CXCL12 [21]。PCR产物在1％的分离的琼脂糖凝胶和0.5染色 μ克/ ml溴化乙锭[21]。

Quantitative PCR was performed with a SYBR Green Master Mix (Elpis Biotech., Daejeon, Korea) in a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). Relative mRNA levels in a sample were based on its threshold cycle in comparison with that of the housekeeping geneβ-actin [22]。

For ELISA, MSCs were cultured for 24 h, culture supernatants were collected, and the concentrations of PGE₂，TGF-β，CCL2, CXCL10, and CXCL12 were measured by using ELISA kits according to the manufacturer’s instructions (Bio-Techne).

2。7。Statistical Analysis

数据表示为 of at least three independentin vitroexperiments performed in triplicates or six mice. To determine statistical significance,values were calculated using one-way ANOVA (GraphPad Software, San Diego, CA, USA).

3.结果

3.1。hMSCs的改善狼疮进展的MRL。FAS^LPR老鼠

We examined the therapeutic activity of hMSCs in MRL.Fas^LPR老鼠。hMSC-treated mice survived at least 24 weeks of age, which was much longer than the control mice (Figure图1（a））。的hMSCs不影响体重（图图1（b））。抗dsDNA的血清水平（图图1（c））and total IgG antibodies (Figure1（d））and proteinuria level (Figure1（e）中）在被hMSC治疗的小鼠中显著低于对照小鼠。正如预期的那样，免疫抑制剂，环磷酰胺，用作阳性对照改善的疾病（图1）。总的来说，我们的数据表明，hMSCs的改善SLE的MRL中的进展.Fas^LPR老鼠。

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图1

hMSCs的改善在MRL SLE发展。FAS ^LPR老鼠。MRL.Fas^LPR小鼠静脉用载体（PBS，对照注射; ），的hMSC（ 细胞/小鼠， ），or cyclophosphamide (50 mg/kg, ）曾经在12周的年龄。存活（a）和体重（B）是每星期测量。抗dsDNA IgG的（c）和总IgG（d）的血清中和水平的蛋白尿（e）中的电平进行了测定。 versus control.

3。2。Effect of hMSCs on Cytokine Gene Expression in the Kidney of MRL.FAS^LPR老鼠

我们分离的总RNA从MRL的肾脏。FAS^LPRmice at 16 weeks of age (4 weeks after hMSC injection or vehicle injection). As additional control, we isolated total RNA from the kidney of MRL.FAS^LPRmice at 9 weeks of age (before hMSC injection). Compared to 9 weeks of age, the expression levels of all the cytokines examined increased at 16 weeks and were decreased by hMSC injection (Figure2）。这些数据表明，hMSCs的可能抑制炎症的免疫细胞，包括T细胞的功能。

图2

的hMSC降低MRL的肾脏细胞因子基因表达。FAS ^LPR老鼠。的hMSC（ 细胞/小鼠， ）were injected once at the age of 12 weeks. Total RNAs were isolated from the kidney at the age of 9 or 16 weeks ( ）。细胞因子的表达水平通过qPCR测量。 versus 9 weeks.^＃ 对的hMSC-未处理的对照（16周）。

3.3。效果从MRL隔离对T的hMSC和B细胞。FAS^LPR老鼠

To examine whether hMSCs directly inhibit mouse T and B cells, we set up a xenogeneic assay system by coculturing hMSCs and immune cells isolated from MRL.Fas^LPR老鼠。hMSCs did not affect the proliferation of or IgM production by LPS-activated B cells (Figure3（a)）。However, hMSCs inhibited the proliferation of or IFN-γproduction by ConA-activated T cells (Figure3（b)）。Nephritic kidney expresses high levels of inflammatory cytokines, including IFN-γ，which could improve the inhibitory capacity of MSCs [2]。Thus, we treated hMSCs with IFN-γin dose- (Figures3（C）and3（e）中）and time-dependent manners (Figures3（d）and3（F））。IFN-γγ-activated hMSCs (IFN-hMSCs) inhibited the IgM production by LPS-activated mouse B cells (Figures3（C）and3（d））and IFN-γproduction by ConA-activated T cells (Figures3（e）中and3（F））。这些数据表明，天真的hMSCs仅抑制T细胞，如果他们被激活干扰素γ，they can inhibit both T and B cells.

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图3

IFN-γγmakes hMSCs capable of inhibiting MRL.FAS ^LPRB cells. (a, b) hMSCs ( cells/well) and MRL.Fas ^LPRmouse B (a) or T (b) cells ( cells/well) were cocultured for 72 h. B cells with lipopolysaccharide (LPS, 1 μg/ml) and T cells were activated with concanavalin A (ConA, 1 μg/ml). Proliferation of B and T cells was measured by the³H-thymidine uptake assay (upper), and the levels of IgM (a) and IFN-γ（B）在培养培养基中积累，通过ELISA（降低）来测定。（C-F）的hMSC用IFN-α预处理γ（3-30 ng/ml) for 7 days (c, e) or with IFN-γ（10 ng/ml) for 1–7 days (d, f). hMSCs ( 细胞/孔）和B细胞（ cells/well) were cocultured for 72 h, and the levels of IgM were measured by ELISA (c, d). hMSCs ( cells/well) and T cells ( cells/well) were cocultured for 72 h, and the levels of IFN-γaccumulated in culture medium were measured by ELISA (e, f). versus control.

3.4。在IDO依赖性方式IFN-的hMSCs抑制小鼠B细胞

Much has been learned about the mechanisms of T cell inhibition by MSCs, whereas little is known about the effects of MSCs on B cells. Thus, we examined the effect of IFN-hMSCs on mouse B cells in greater detail. To assess whether IFN-hMSCs inhibit mouse B cells in a soluble factor- or contact-dependent manner, we used a transwell assay. When we added IFN-hMSCs and mouse B cells to the lower wells, thereby allowing cell–cell contact, IFN-hMSCs strongly inhibited IgM production by B cells (Figure4（a)）。当我们加入IFN-的hMSCs到上部井和小鼠B细胞下层孔，从而防止直接的细胞 - 细胞接触，抑制较弱（图4（a)）。These data imply that IFN-hMSCs inhibit mouse B cell functions in both contact- and soluble factor-dependent manners. Naïve hMSCs expressed the immunosuppressive soluble factors cyclooxygenase-2 (COX-2), PGE₂和TGF-ββ和IFN-的hMSCs另外表达IDO（图4（b)-4（d）），这表明IFN-的hMSCs可能使用IDO B细胞的抑制作用。IFN-γγincreased IDO expression by phosphorylating STAT1 in a time-dependent manner (Figure4（e）中）;STAT1抑制剂氟达拉滨抑制STAT1磷酸化和IDO表达（图4（F））。The ability of IFN-hMSCs to inhibit B cells was abolished by the transfection of IFN-hMSCs with IDO siRNA (Figure4（G））and treatment with IDO inhibitor 1MT (Figure4（H）），氟达拉滨或（图4（I））。These data suggest that IFN-hMSCs inhibit mouse B cell functions in an IDO-dependent manner.

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图4

IFN-的hMSCs抑制MRL。FAS ^LPRB cells in an indoleamine 2,3-dioxygenase-dependent manner. (a) IFN-γ活化的hMSC（IFN-的hMSCs; 细胞/孔）加入到上部（U）或降低（L）的transwell板和MRL的孔.Fas ^LPR小鼠的B细胞（ cells/well) to the L wells. After incubation with LPS for 72 h, the level of IgM accumulated in culture medium was measured by ELISA. (b) Total RNA was isolated from IFN-hMSCs, and the expression levels of COX-2, TGF-β，and IDO were analyzed by RT-PCR. (c) IDO protein level in IFN-hMSCs analyzed by western blotting. (d) Levels of PGE₂and TGF-βaccumulated in IFN-hMSC culture medium over 24 h measured by ELISA (d). (e, f) hMSCs were treated with IFN-γ（10 ng/ml) for 7 days and the levels of IDO, STAT1, and p-STAT1 were analyzed by western blotting (e). In (f), the cells were treated with the STAT1 inhibitor fludarabine (10 nM) for 7 days. (g–i) IFN-hMSCs were transfected with IDO siRNA for 48 h (g) or treated with the IDO inhibitor 1-methyltryptophan (1MT, 100 μM) for 7 days (h) or fludarabine (10 nM) for 7 days (i). The level of IgM accumulated in culture medium was measured by ELISA. （ ）。NS: not significant. (b, c, e, and f) Band areas were analyzed by ImageJ software (NIH, Bethesda, MD, USA), and data were presented as ratios versus UN (b, c), day 0 (e), and IFN/fludarabine-untreated group (f) ( ）。

3.5。IFN-的hMSCs抑制小鼠B细胞在CXCL10依赖性方式

Next, we examined how IFN-hMSCs contact mouse B cells. IFN-hMSCs adhere to dishes and mouse B cells grow in suspension, suggesting that B cells might migrate toward IFN-hMSCs after sensing IFN-hMSC-derived chemokines. Thus, we assessed the profiles of chemokines expressed by IFN-hMSCs and chemokine receptors in mouse B cells. Naïve hMSCs expressed CCL2 and CXCL12, and IFN-hMSCs additionally expressed CXCL10, suggesting a potential role of CXCL10 in MSC–B cell interactions (Figures五（a)and五（b)）。为了证明这一假设，我们撞倒CCL2，CXCL10，CXCL12或用各自的siRNA（图五（C））。IFN-γhMSCs transfected with CCL2 or CXCL12 siRNA, but not IFN-hMSCs transfected with CXCL10 siRNA, induced mouse B cell migration in transwell assay (Figure五（d））。我们进行了延时成像来评估在单细胞水平的小鼠B细胞向IFN-的hMSC迁移。We placed IFN-hMSCs on the left side of an imaging chamber and mouse B cells on the right side and acquired images every 2 min for 12 h (Supplementary Video1）。Representative images and the number of mouse B cells passing through a selected area (white box) are shown in Figure五（e）中。IFN-γhMSCs transfected with negative-control or CCL2 siRNA induced B cell migration, whereas CXCL10 siRNA-transfected IFN-hMSCs did not (Figure五（e）中）。总体而言，这些数据表明，IFN-的hMSCs吸引用于在CXCL10依赖性方式接触依赖性抑制小鼠B细胞。

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图5

IFN-的hMSCs抑制MRL。FAS ^LPRB cells in a CXCL10-dependent manner. (a, b) Expression levels of chemokines in MSCs measured by RT-PCR (a) and ELISA (b). UN: untreated hMSCs. IFN: IFN-treated hMSCs. (c–e) IFN-γ活化的hMSC（IFN-的hMSCs）与阴性对照，CCL2，CXCL10，CXCL12或siRNA的转染。表达水平分析通过RT-PCR（c）中。IFN-的hMSCs（ 细胞/孔）加入到下层孔和MRL.Fas ^LPR小鼠的B细胞（ cells/well) to the upper wells of transwell plates with a 5 μ米插入。After 1.5 h, the number of B cells migrating to the lower well was determined (d). For time-lapse imaging, IFN-hMSCs (70 μl of cells/ml) were seeded into the left chamber and B cells (70 μl of 细胞/ ml）到培养嵌件的右腔室μ-Dish^3五mmculture dishes. Images were acquired every 2 min for 12 h ( ）。Representative photos are shown. The number of B cells passing through the white boxes is shown on the right (e). 。

4。Discussion

MRL。FAS^LPR小鼠已被广泛用于评估对SLE的hMSCs的临床前功效。这些小鼠缺乏FASgene and spontaneously develop SLE-like symptoms, including an increase in anti-dsDNA antibodies in blood, and develop severe nephritis [23]。Similar to SLE patients, the symptom severity in MRL.Fas^LPR小鼠取决于性别：雌性小鼠17周的平均年龄死亡，22周男性。一些临床前研究已经证实，hMSCs的改善在MRL SLE进展.Fas^LPR老鼠。Lee等人。[14] showed that the transfer of hMSCs increased MRL.FAS^LPR小鼠存活，在肾脏降低T细胞浸润，和降低的T细胞细胞因子的表达。Lee等人。[24]还表明，与泼尼松或霉酚酸酯组合的hMSC具有比在MRL单一疗法更好的治疗效果。FAS^LPR老鼠;下面读数是在本研究中使用：生存期的延长，降低抗dsDNA和血清总IgG水平，降低细胞因子基因表达中的脾细胞，并减少炎性细胞浸润入肾。刘等人。[2五] showed that hMSCs reduced proteinuria level, serum anti-dsDNA antibody level, and renal damage in MRL.FAS^LPR老鼠。In agreement with these previous data, our data confirm that xenogeneic hMSCs ameliorate SLE progression in MRL.FAS^LPR老鼠。显示的hMSC注射的小鼠抗体产生和降低细胞因子的表达，这表明免疫细胞，包括T和B细胞的抑制降低。总体而言，临床前研究上面引用和我们目前的研究表明，在MRL是有效的hMSCs改善狼疮进展。FAS^LPR老鼠。

Although human or mouse MSCs have been reported to inhibit species-matched T and B cells [26-28]，它一直是不清楚的hMSC是否将直接影响异种T和B细胞的功能中MRL。FAS^LPR老鼠。我们证明这天真hMSCs抑制use T cells but not mouse B cells. However, hMSCs might be able to inhibit B cells in mice as the disease progresses. The previous studies [26-28] and our data demonstrate that the nephritic kidney of MRL.FAS^LPRmice expresses high levels of many inflammatory cytokines, including IFN-γ。IFN-γγ上调IDO的通过的hMSC [表达式2，29，三十]。IDO是一个功能强大的免疫抑制酶催化在L-色氨酸分解代谢的第一步[31]。IDO耗尽L-色氨酸和犬尿氨酸产生，这可能会抑制B和T细胞[31]。IFN-的hMSCs抑制T的B细胞增殖和B细胞和IgM产生在IDO依赖性方式[29，三十]。Our data confirm that IFN-hMSCs inhibit mouse B cell functions in an IDO-dependent manner. Here, we also show that IFN-γ通过增加的hMSCs CXCL10生产。虽然IFN-的hMSCs产生若干趋化因子，包括CCL2，CXCL10，CXCL12和，它们仅使用CXCL10吸引小鼠的B细胞。正如我们先前报道，hMSCs的使用CCL2吸引小鼠T细胞14]。总之，这些数据表明，吸引的hMSCs小鼠T和B细胞接触抑制通过使用不同的趋化因子。总之，它是有据可查的，干细胞抑制T细胞功能在两个非接触和可溶性因子依赖性方式[14]。我们目前的研究表明，IFN-的hMSCs抑制B细胞以同样的方式。

五。Conclusion

Our study has an inevitable limitation: our data might not be clinically meaningful, since we used hMSCs as effector cells and mouse T and B cells as target cells. However, our data provide important preclinical information on how hMSCs ameliorate SLE progression in lupus-prone MRL.FAS^LPR老鼠。在注射到小鼠体内后的早期阶段，幼稚的hMSCs抑制小鼠T细胞，而不是小鼠的B细胞。以后，如果的hMSCs被激活干扰素γin mice, they can inhibit mouse B cells in a CXCL10- and IDO-dependent manner. Our data might help to understand how hMSCs ameliorate SLE progression in MRL.FAS^LPRmice, which are widely used to evaluate the preclinical efficacy of hMSCs against SLE.

Data Availability

The data used to support the findings of this study are included within the article.

Conflicts of Interest

被披露没有潜在的利益冲突。

作者的贡献

香港庆利，恩扬金，和亨淑金同等贡献这项工作。

致谢

This study was supported by grants funded by the Korea Government (NRF-2017R1A5A2015541 and NRF-2017M3A9B4050336).

Supplementary Materials

Supplementary Video 1: migration of mouse B cells to IFN-hMSCs. IFN-hMSCs transfected with negative control siRNA (a) and CXCL10 siRNA (b) were loaded in the left chamber. Mouse B cells were loaded in the right chamber. B cell migration was imaged every 2 min for 12 h. Related to Figure五（e）中。（Supplementary Materials）

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Copyright

Copyright © 2020 Hong Kyung Lee et al. This is an open access article distributed under the知识共享署名许可,它允许无限制的使用、分配,一个d reproduction in any medium, provided the original work is properly cited.