人脐带间充质干细胞分化为胰岛素分泌细胞的免疫原性变化
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摘要
背景:人脐带间充质干细胞具有低免疫原性,而人脐带间充质干细胞诱导分化的胰岛素分泌细胞是否保持低免疫原性尚不清楚。目的:探讨人脐带间充质干细胞向胰岛素分泌细胞诱导分化后在体外的免疫原性变化及移植到宿主后微环境对免疫原性的影响。方法:(1)采用经典改良方案诱导人脐带间充质干细胞分化为胰岛素分泌细胞,流式细胞术检测胰岛素分泌细胞免疫表型及细胞毒性实验中胰岛素分泌细胞的凋亡率;(2)CCK-8检测单向混合淋巴细胞实验中外周血单个核细胞的增殖能力;(3)小鼠腹腔及左肾包膜移植胰岛素分泌细胞后,流式细胞术及免疫组织化学检测移植部位的免疫细胞浸润情况。结果与结论:(1)人脐带间充质干细胞分化的胰岛素分泌细胞高表达HLA-ABC,低表达HLA-DR,CD40和CD80;(2)细胞毒性实验中胰岛素分泌细胞的凋亡率随预致敏脾细胞的增多而升高;(3)单向混合淋巴细胞实验中效靶比为10︰1,50︰1时胰岛素分泌细胞能抑制外周血单个核细胞增殖;(4)腹腔及左肾包膜移植胰岛素分泌细胞后移植部位淋巴细胞增多;(5)结果表明,人脐带间充质干细胞诱导分化的胰岛素分泌细胞在体外保持低免疫原性,但移植到体内后由于微环境的变化而具有一定免疫原性。
BACKGROUND: Human umbilical cord mesenchymal stem cells(h UC-MSCs) have low immunogenicity and it is unclear whether insulin producing cells(IPCs) that differentiate from hUC-MSCs have low immunogenicity. OBJECTIVE: To investigate the immunogenicity of IPCs differentiating from hUC-MSCs in vitro and after IPCs transplantation into the host. METHODS:(1) The hUC-MSCs were induced to differentiate into IPCs according to the modified scheme. Flow cytometry assay was used to detect the immunophenotype and apoptotic rate of IPCs in a cytotoxicity test.(2) Cell counting kit-8 was used to detect the proliferative capacity of human peripheral blood mononuclear cells in the one-way mixed lymphocyte assay.(3) The IPCs were then transplanted into the abdominal cavity and left renal capsule of mice, and then the infiltration of immune cells was detected by flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: The IPCs highly expressed HLA-ABC and lowly expressed HLA-DR, CD40 and CD80. The apoptosis rate of IPCs increased with the increase of pre-sensitized splenocytes in the cytotoxicity test. In the one-way mixed lymphocyte assay, IPCs inhibited the proliferation of human peripheral blood mononuclear cells when the target ratio was 10:1 and 50:1. After IPCs transplantation, the number of lymphocytes was increased in the transplanted site. In summary, our results show that IPCs that differentiate from hUC-MSCs maintain low immunogenicity in vitro, but have some immunogenicity after transplantation into the host due to microenvironment changes.
BACKGROUND: Human umbilical cord mesenchymal stem cells(h UC-MSCs) have low immunogenicity and it is unclear whether insulin producing cells(IPCs) that differentiate from hUC-MSCs have low immunogenicity. OBJECTIVE: To investigate the immunogenicity of IPCs differentiating from hUC-MSCs in vitro and after IPCs transplantation into the host. METHODS:(1) The hUC-MSCs were induced to differentiate into IPCs according to the modified scheme. Flow cytometry assay was used to detect the immunophenotype and apoptotic rate of IPCs in a cytotoxicity test.(2) Cell counting kit-8 was used to detect the proliferative capacity of human peripheral blood mononuclear cells in the one-way mixed lymphocyte assay.(3) The IPCs were then transplanted into the abdominal cavity and left renal capsule of mice, and then the infiltration of immune cells was detected by flow cytometry and immunohistochemistry. RESULTS AND CONCLUSION: The IPCs highly expressed HLA-ABC and lowly expressed HLA-DR, CD40 and CD80. The apoptosis rate of IPCs increased with the increase of pre-sensitized splenocytes in the cytotoxicity test. In the one-way mixed lymphocyte assay, IPCs inhibited the proliferation of human peripheral blood mononuclear cells when the target ratio was 10:1 and 50:1. After IPCs transplantation, the number of lymphocytes was increased in the transplanted site. In summary, our results show that IPCs that differentiate from hUC-MSCs maintain low immunogenicity in vitro, but have some immunogenicity after transplantation into the host due to microenvironment changes.
引文
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[23]Yang D,Lin H,Liu G.Immunogenicity of human umbilical cord blood derived mesenchymal stem cells after osteogenic induction.Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi.2014;28(6):752-757.
[24]Li Q,Zhang H,Yu L,et al.Down-regulation of Notch signaling pathway reverses the Th1/Th2 imbalance in tuberculosis patients.Int Immunopharmacol.2018;54:24-32.
[25]Qian LJ,Kang SM,Xie JL,et al.Early-life gut microbial colonization shapes Th1/Th2 balance in asthma model in BALB/c mice.BMC Microbiol.2017;17(1):135.
[26]Takahashi N,Saitoh T,Gotoh N,et al.The cytokine polymorphisms affecting Th1/Th2 increase the susceptibility to,and severity of,chronic ITP.BMC Immunol.2017;18(1):26.
[2]Ahearn AJ,Parekh JR,Posselt AM.Islet transplantation for Type 1 diabetes:where are we now.Expert Rev Clin Immunol.2015;11(1):59-68.
[3]Zhao K,Lou R,Huang F,et al.Immunomodulation effects of mesenchymal stromal cells on acute graft-versus-host disease after hematopoietic stem cell transplantation.Biol Blood Marrow Transplant.2015;21(1):97-104.
[4]Berglund AK,Fisher MB,Cameron KA,et al.Transforming Growth Factor-β2 Downregulates Major Histocompatibility Complex(MHC)I and MHC II Surface Expression on Equine Bone Marrow-Derived Mesenchymal Stem Cells Without Altering Other Phenotypic Cell Surface Markers.Front Vet Sci.2017;4:84.
[5]Tan K,Zheng K,Li D,et al.Impact of adipose tissue or umbilical cord derived mesenchymal stem cells on the immunogenicity of human cord blood derived endothelial progenitor cells.PLo S One.2017;12(5):e0178624.
[6]Maria AT,Maumus M,Le Quellec A,et al.Adipose-Derived Mesenchymal Stem Cells in Autoimmune Disorders:State of the Art and Perspectives for Systemic Sclerosis.Clin Rev Allergy Immunol.2017;52(2):234-259.
[7]Gu LH,Zhang TT,Li Y,et al.Immunogenicity of allogeneic mesenchymal stem cells transplanted via different routes in diabetic rats.Cell Mol Immunol.2015;12(4):444-455.
[8]Mohammadi Ayenehdeh J,Niknam B,Rasouli S,et al.Immunomodulatory and protective effects of adipose tissue-derived mesenchymal stem cells in an allograft islet composite transplantation for experimental autoimmune type1 diabetes.Immunol Lett.2017;188:21-31.
[9]Lee HJ,Kang KS,Kang SY,et al.Immunologic properties of differentiated and undifferentiated mesenchymal stem cells derived from umbilical cord blood.J Vet Sci.2016;17(3):289-297.
[10]Ciapetti G,Granchi D,Fotia C,et al.Effects of hypoxia on osteogenic differentiation of mesenchymal stromal cells used as a cell therapy for avascular necrosis of the femoral head.Cytotherapy.2016;18(9):1087-1099.
[11]Helal MA,Shaheen NE,Abu Zahra FA.Immunomodulatory capacity of the local mesenchymal stem cells transplantation after severe skeletal muscle injury in female rats.Immunopharmacol Immunotoxicol.2016:1-9.[Epub ahead of print]
[12]Fu X,Yang H,Zhang H,et al.Improved osteogenesis and upregulated immunogenicity in human placenta-derived mesenchymal stem cells primed with osteogenic induction medium.Stem Cell Res Ther.2016;7(1):138.
[13]Yang R,Yu T,Zhou Y.Interplay between craniofacial stem cells and immune stimulus.Stem Cell Res Ther.2017;8(1):147.
[14]Dhingra S,Li P,Huang XP,et al.Preserving prostaglandin E2level prevents rejection of implanted allogeneic mesenchymal stem cells and restores postinfarction ventricular function.Circulation.2013;128(11 Suppl 1):S69-78.
[15]Chen T,Wang Y,Gu L,et al.Role of INGAP-pp in the differentiation of h UCMSCs into insulin producing cells.Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi.2013;29(2):141-145.
[16]Foroutan T,Farhadi A,Abroun S,et al.Adipose Derived Stem Cells Affect mi R-145 and p53 Expressions of Co-Cultured Hematopoietic Stem Cells.Cell J.2018;19(4):654-659.
[17]Sivanathan KN,Gronthos S,Grey ST,et al.Immunodepletion and Hypoxia Preconditioning of Mouse Compact Bone Cells as a Novel Protocol to Isolate Highly Immunosuppressive Mesenchymal Stem Cells.Stem Cells Dev.2017;26(7):512-527.
[18]Lee S,Jeong S,Lee C,et al.Mesenchymal Stem Cells Derived from Human Exocrine Pancreas Spontaneously Express Pancreas Progenitor-Cell Markers in a Cell-Passage-Dependent Manner.Stem Cells Int.2016;2016:2142646.
[19]Modiano JF,Lindborg BA,Mc Elmurry RT,et al.Mesenchymal stromal cells inhibit murine syngeneic anti-tumor immune responses by attenuating inflammation and reorganizing the tumor microenvironment.Cancer Immunol Immunother.2015;64(11):1449-1460.
[20]Hoogduijn MJ.Are mesenchymal stromal cells immune cells.Arthritis Res Ther.2015;17:88.
[21]Glenn JD,Whartenby KA.Mesenchymal stem cells:Emerging mechanisms of immunomodulation and therapy.World J Stem Cells.2014;6(5):526-539.
[22]Yang JF,Cao HC,Pan QL,et al.Mesenchymal stem cells from the human umbilical cord ameliorate fulminant hepatic failure and increase survival in mice.Hepatobiliary Pancreat Dis Int.2015;14(2):186-193.
[23]Yang D,Lin H,Liu G.Immunogenicity of human umbilical cord blood derived mesenchymal stem cells after osteogenic induction.Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi.2014;28(6):752-757.
[24]Li Q,Zhang H,Yu L,et al.Down-regulation of Notch signaling pathway reverses the Th1/Th2 imbalance in tuberculosis patients.Int Immunopharmacol.2018;54:24-32.
[25]Qian LJ,Kang SM,Xie JL,et al.Early-life gut microbial colonization shapes Th1/Th2 balance in asthma model in BALB/c mice.BMC Microbiol.2017;17(1):135.
[26]Takahashi N,Saitoh T,Gotoh N,et al.The cytokine polymorphisms affecting Th1/Th2 increase the susceptibility to,and severity of,chronic ITP.BMC Immunol.2017;18(1):26.