Na Zhao

Cell fusion between gastric epithelial cells and mesenchymal stem cells results in epithelial-to-mesenchymal transition and malignant transformation

BackgroundThe discovery of cancer stem cells and tumor heterogeneity prompted the exploration of additional mechanisms aside from genetic mutations for carcinogenesis and cancer progression. The aim of the present study was to investigate the effect of cell fusion between mesenchymal stem cells and the gastric epithelial cells in tumorigenesis.MethodsCell fusion between cord blood mesenchymal stem cells and human gastric epithelial cells was performed in vitro. Cell scratch and transwell assays were performed to determine migration and invasion abilities of the hybrids. The expressions of epithelial-mesenchymal transition-related proteins and genes were analyzed by immunocytochemistry and real time quantitative PCR. Tumorigenesis of the hybrids was evaluated through in vivo inoculation in nude mice.ResultsHybrids expressed the phenotypes of both donor cells. Aneuploidy was observed in 84.1% of cells. The hybrids showed increased proliferation, migration and invasion abilities compared with the parental cells. In addition, the expression of N-cadherin and vimentin in the hybrids was significantly higher than that of the epithelial cells, and the mRNA expression of the epithelial-mesenchymal transition-related genes, Twist and Slug, in the hybrids was also increased compared with that of the parental epithelial cells. Furthermore, the hybrids formed masses of epithelial origin with glandular structures in BALB/c nude mice.ConclusionsThese findings suggest that cell fusion between gastric epithelial cells and mesenchymal stem cells may result in epithelial to mesenchymal transition and malignant transformation.

Inhibiting cardiac allograft rejection with interleukin-35 therapy combined with decitabine treatment in mice ☆ ☆☆

BACKGROUNDnRegulatory T (Treg) cells play a pivotal role in the maintenance of transplantation tolerance. It is of great interest to induce allograft tolerance mediated by regulatory CD4(+)CD25(+) T cells.nnnMETHODSnHere we investigated the effect of hydrodynamic IL-35-expressing plasmid injection in combination with a methyltransferase inhibitor (decitabine) on immune function and transplantation tolerance in mice.nnnRESULTSnWe showed that IL-35 and decitabine stimulated the proliferation of CD4(+)CD25(+) Tregs and suppressed CD8(+) T cell proliferation in an allogenic mixed lymphocyte culture in vitro. IL-35 gene therapy and decitabine administration prolonged the survival of the transplanted heart in the heterotopic abdominal heart transplantation model in mice.nnnCONCLUSIONSnThe possible mechanism through which IL-35 and decitabine treatment increased the survival of graft tissues is to enhance the proliferation of CD4(+)CD25(+) Treg cells and suppress the generation and function of effector T cells. Thus, IL-35 gene therapy combined with decitabine provides a novel approach to induce transplantation tolerance.

Interleukin-15 gene therapy and the mammalian target of rapamycin inhibitor everolimus inhibit the growth of metastatic breast cancer.

Novel methods to control and treat metastatic breast cancer are needed. Interleukin (IL)‐15 is a promising cytokine for cancer immunotherapy, and everolimus is an orally administered mammalian target of rapamycin (mTOR) inhibitor, which is already approved for cancer treatment. In the present study, we investigated the efficacy of IL‐15 gene therapy and explored the possibility of combining IL‐15 therapy with everolimus to treat metastatic breast cancer.

Mesenchymal Stem Cells Overexpressing Interleukin-35 Propagate Immunosuppressive Effects in Mice

To explore generation of interleukin (IL)‐35‐expressing mouse adipocyte‐derived mesenchymal stem cells (Ad‐MSCs) using lentiviral vector and their potential immunosuppressive effects in mice. Ad‐MSCs were isolated and cultured in vitro and transfected with a lentivirus vector for overexpression of the therapeutic murine IL‐35 gene. IL‐35 expression in transfected MSCs (IL‐35‐MSCs) was quantified by enzyme‐linked immunosorbent assay (ELISA). The lymphocytes subsets after one‐way mixed lymphocyte culture and in vivo intravenous transplantation were analysed by flow cytometry to evaluate the immunosuppressive effects of IL‐35‐MSCs. ELISA was performed to examine IL‐10, IL‐17A and IL‐35 expression in lymphocyte culture. Mouse Ad‐MSCs were isolated and cultured. IL‐35 was expressed in the MSC supernatant and serum after IL‐35 transduction into Ad‐MSCs by lentiviral vector transfection in vitro and in vivo. The percentage of CD4+ CD25+ T regulatory (Treg) cells in mice treated with IL‐35‐MSCs significantly increased. IL‐35‐MSCs upregulated the CD4+ CD25+ Treg cells in the allogeneic mixed lymphocyte reaction system, and lowered the percentage of CD4+ T cells compared with the other two control groups (P < 0.01). IL‐17A expression significantly decreased and IL‐10 expression significantly increased in IL‐35‐MSCs and MSCs when compared by ELISA to the control groups (P < 0.01). IL‐35‐transduced Ad‐MSCs in vivo can enhance proliferation of CD4+ CD25+ Treg cells and suppress the function of effector T cells such as T helper (Th) 1, Th2 and Th17 cells and may reduce the development of allograft rejection. Our data suggest that transduced Ad‐MSCs overexpressing IL‐35 may provide a useful approach for basic research on cell‐based immunotolerance therapy for inducing transplantation tolerance.

Interleukin-35 Gene-Modified Mesenchymal Stem Cells Protect Concanavalin A–Induced Fulminant Hepatitis by Decreasing the Interferon Gamma Level

Interleukin 35 (IL-35) is a relatively newly identified cytokine required for the regulatory and suppressive functions of regulatory T cells (Treg), playing an important role in the prevention of autoimmune diseases. This study used mesenchymal stem cells (MSCs) as the gene-delivery vehicles for IL-35 gene therapy and investigated their protective effects in Concanavalin A (Con A)-induced autoimmune hepatitis. Results showed that IL-35 gene modified MSCs (IL-35-MSCs) can specifically migrate to the injured liver tissues and significantly narrow the necrosis areas of injured livers. IL-35-MSCs prevented hepatocyte apoptosis by reducing the FASL expression by mononuclear cells. Although MSC treatment can alleviate liver injury to some extent, IL-35-MSCs showed a stronger protective effect, which means some novel mechanisms exist. The results show that IL-35-MSCs could decrease the level of interferon gamma secreted by liver mononuclear cells through the JAK1-STAT1/STAT4 signal pathway. In summary, this study thus demonstrates a novel and efficient treatment for Con A-induced fulminant hepatitis through negatively regulating the secretion of interferon gamma, thus providing a novel therapeutic approach for this devastating liver disease.

Mesenchymal stem cell expression of interleukin-35 protects against ulcerative colitis by suppressing mucosal immune responses

BACKGROUNDnInterleukin-35 (IL-35) has recently been identified as an immunosuppressive cytokine that has been used as a potential therapy for chronic inflammatory and autoimmune diseases. However, there remains a paucity of data regarding its potential benefits after integration into mesenchymal stem cells (MSCs).nnnMETHODSnWe used a dextran sulfate sodium (DSS)-induced colitis mice model and treated them with IL-35-MSCs, MSCs or saline. The body weight was recorded daily and inflammatory processes were determined. Cytokine secretion by lamina propria lymphocytes (LPLs) and percentage of regulatory T cells (Tregs) were also measured.nnnRESULTSnThe data showed that mice in the two treated groups recovered their body weight more rapidly than mice treated with saline in the later stage of colitis. The colon lengths of IL-35-MSC-treated mice were markedly longer than those in the other two groups and the inflammation reduced significantly. Furthermore, the percentage of Foxp3u2009+u2009Tregs increased significantly and the level of proinflammatory cytokines produced by LPLs decreased significantly in the IL-35-MSC-treated group.nnnDISCUSSIONnThe results demonstrate that IL-35-MSCs could ameliorate ulcerative colitis by down-regulating the expression of pro-inflammatory cytokines.

Cytokine profiles in papillary thyroid carcinoma, with or without Hashimoto’s thyroiditis:

The phenomenon that papillary thyroid carcinoma (PTC) is often accompanied by Hashimoto’s thyroiditis (HT) raised interest in further study of immune changes caused by thyroid autoimmunity. We aimed to characterize cytokine profiles in the peripheral blood of patients with PTC with or without HT, compared with nodular goiter (NG) and healthy control (HC) patients, in order to determine the autoimmunity-related differences among these groups. A total of 50 PTC patients were divided into two groups, according to whether or not concurrent HT existed. A total of 20 NG patients and 20 HC subjects were also included as controls. All PTC patients and NG patients who underwent surgical thyroidectomy and pathology examination were included. The serum levels of four cytokines, including interferon gamma (IFN-γ), interleukin (IL)-17, IL-10, and IL-35 were measured using AimPlex bead–based immunoassays. Peripheral IFN-γ and IL-35 increased significantly in PTC patients with concurrent HT. IL-10 and IL-35 increased significantly in three groups (PTC with or without HT and NG patients), while IFN-γ increased only in the two PTC groups. IL-17 was highest in the HC group, but significantly lower in the other three groups. IL-17, IL-10, and IL-35 may be involved in tumor onset and progression. Moreover, IFN-γ and IL-35 may play vital roles in the concurrence of PTC and HT, which could warrant further exploration.

Mesenchymal stem cells overexpressing IL-35: a novel immunosuppressive strategy and therapeutic target for inducing transplant tolerance

Inducing donor-specific immunological tolerance, which avoids the complications of long-term immunosuppression, is an important goal in organ transplantation. Interleukin-35 (IL-35), a cytokine identified in 2007, is mainly secreted by regulatory T cells (Tregs) and is essential for Tregs to exert their maximal immunoregulatory activity in vitro and in vivo. A growing number of studies show that IL-35 plays an important role in autoimmune diseases and infectious diseases. Recent research has shown that IL-35 could effectively alleviate allograft rejection and has the potential to be a novel therapeutic strategy for graft rejection. With increasing study of immunoregulation, cell-based therapy has become a novel approach to attenuate rejection after transplantation. Mesenchymal stem cells (MSCs), which exhibit important properties of multilineage differentiation, tissue repair, and immunoregulation, have recently emerged as attractive candidates for cell-based therapeutics, especially in transplantation. Accumulating evidence demonstrates that the therapeutic abilities of MSCs can be amplified by gene modification. Therefore, researchers have constructed IL-35 gene-modified MSCs and explored their functions and mechanisms in some disease models. In this review, we discuss the potential tolerance-inducing effects of MSCs in transplantation and briefly introduce the immunoregulatory functions of the IL-35 gene-modified MSCs.

IL-35 Gene Modified Mesenchymal Stem Cells Protect the Con A Induced Fulminant Hepatitis Through Decreasing the IFN-g level

Interleukin 35 (IL-35) is a relatively newly identified cytokine required for the regulatory and suppressive functions of regulatory T cells (Treg), playing an important role in the prevention of autoimmune diseases. This study used mesenchymal stem cells (MSCs) as the gene-delivery vehicles for IL-35 gene therapy and investigated their protective effects in Concanavalin A (Con A)-induced autoimmune hepatitis. Results showed that IL-35 gene modified MSCs (IL-35-MSCs) can specifically migrate to the injured liver tissues and significantly narrow the necrosis areas of injured livers. IL-35-MSCs prevented hepatocyte apoptosis by reducing the FASL expression by mononuclear cells. Although MSC treatment can alleviate liver injury to some extent, IL-35-MSCs showed a stronger protective effect, which means some novel mechanisms exist. The results show that IL-35-MSCs could decrease the level of interferon gamma secreted by liver mononuclear cells through the JAK1-STAT1/STAT4 signal pathway. In summary, this study thus demonstrates a novel and efficient treatment for Con A-induced fulminant hepatitis through negatively regulating the secretion of interferon gamma, thus providing a novel therapeutic approach for this devastating liver disease.

Effects of Platelet-Derived Endothelial Cell Growth Factor and Doppler Perfusion Index in Patients with Colorectal Hepatic Metastases.

Background: We aimed to find out if there is a correlation between Doppler perfusion index (DPI) and platelet- derived endothelial cell growth factor (PD-ECGF), which is an angiogenic factor with angiopoietic function, in patients with colorectal carcinoma. Methods: 50 colorectal carcinoma patients (22 cases with liver metastases, 28 cases without liver metastases) and 50 healthy controls were assessed with Doppler ultrasound as a preoperative evaluation. PD-ECGF expression in postoperative specimens of the 50 cases with colorectal carcinoma was assayed by immunohistochemistry and real-time polymerase chain reaction methods. Results: The mean DPI value was 0.29 ± 0.05 in patients suffering from colon cancer with hepatic metastases and 0.12 ± 0.03 in the healthy control group. The DPI value was significantly higher in patients with liver metastases (p < 0.05). PD-ECGF expression in patients with colorectal liver metastases was significantly higher than that in the group without liver metastases (p < 0.05). A positive correlation was found between DPI value and PD-ECGF expression in patients with liver metastases (p < 0.05). Conclusions: DPI and PD-ECGF may be valuable factors when screening hepatic metastases in patients with colorectal cancer and serve as practical measurements in postoperative follow-up.