人胎盘间充质干细胞CD200~+亚群细胞的免疫特性
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摘要
背景:在不同的研究中,间充质干细胞表现出相悖的免疫特性,这可能是因为间充质干细胞是一群复合细胞,不同的亚群细胞表现出不同的免疫特性。目的:从人胎盘间充质干细胞中分选获得CD200+亚群细胞,探讨该亚群细胞的免疫特性。方法:采用流式细胞方法从人胎盘间充质干细胞中分选出CD200~+亚群细胞,对其进行细胞表型、多向分化潜能鉴定;ELISA方法检测CD200~+亚群细胞的白细胞介素6、白细胞介素8因子分泌量;混合淋巴细胞共培养体系检测CD200~+亚群细胞的免疫调节功能。结果与结论:(1)流式细胞术成功从人胎盘间充质干细胞中分选获得CD200~+亚群细胞,该亚群细胞高表达CD44、CD73、CD105和CD90,而CD34、CD45、CD80和CD86为阴性,具有典型的间充质干细胞表面标志,可诱导分化为脂肪细胞和成骨细胞,分泌高水平的白细胞介素6和白细胞介素8;(2)淋巴细胞共培养体系中,与人胎盘间充质干细胞和CD200-亚群细胞相比,CD200~+亚群细胞能够显著抑制被活化的淋巴细胞增殖,下调干扰素γ、白细胞介素2水平,而白细胞介素10水平明显上调;(3)该研究发现,从人胎盘间充质干细胞中分离获得的CD200~+亚群细胞具有优于胎盘间充质干细胞和CD200~-亚群细胞的免疫调节功能,为其临床应用提供了一定的理论依据。
BACKGROUND: In different studies, mesenchymal stem cells exhibit contradictory immune characteristics. This may be because mesenchymal stem cells are a group of complex cells and different subgroups exhibit different immune characteristics.OBJECTIVE: To isolate and identify a sub-population of placenta-derived mesenchymal stem cells with high positive surface antigen of CD200, and to investigate the immune characteristics of this sub-population. METHODS: Flow cytometry was utilized to sort CD200~+ placenta-derived mesenchymal stem cells. Phenotypic characterization and multilineage differentiation of the sorted cells were determined. Cytokine secretion, involving interleukin-6 and interleukin-8, was measured by ELISA. Mixed lymphocyte proliferation assay was used to determine the immune characteristics of CD200~+ placenta-derived mesenchymal stem cells. RESULTS AND CONCLUSION: The CD200~+ placenta-derived mesenchymal stem cells exhibited homogeneous spindle shape, and highly expressed CD44, CD73, CD90 and CD105, negatively expressed CD34, CD45, CD80 and CD86. After induced differentiation, the cells were positively stained by oil red O staining and alizarlin red staining, which were the markers of adipocytes and osteoblasts. The results of ELISA indicated that the expression level of interleukin-6 and interleukin-8 in the CD200~+ placenta-derived mesenchymal stem cells was strikingly higher than placenta-derived mesenchymal stem cells and CD200~-placenta-derived mesenchymal stem cells. Compared with normal and CD200~-placenta-derived mesenchymal stem cells, CD200~+ placenta-derived mesenchymal stem cells inhibited T cell proliferation significantly, lowered the levels of interferon-γ and interleukin-2, and upregulated the level of interleukin-10. We found that CD200~+ placenta-derived mesenchymal stem cells have better immunoregulatory capability compared with normal and CD200~-placenta-derived mesenchymal stem cells, which may provide an ideal theoretical basis for clinical use.
BACKGROUND: In different studies, mesenchymal stem cells exhibit contradictory immune characteristics. This may be because mesenchymal stem cells are a group of complex cells and different subgroups exhibit different immune characteristics.OBJECTIVE: To isolate and identify a sub-population of placenta-derived mesenchymal stem cells with high positive surface antigen of CD200, and to investigate the immune characteristics of this sub-population. METHODS: Flow cytometry was utilized to sort CD200~+ placenta-derived mesenchymal stem cells. Phenotypic characterization and multilineage differentiation of the sorted cells were determined. Cytokine secretion, involving interleukin-6 and interleukin-8, was measured by ELISA. Mixed lymphocyte proliferation assay was used to determine the immune characteristics of CD200~+ placenta-derived mesenchymal stem cells. RESULTS AND CONCLUSION: The CD200~+ placenta-derived mesenchymal stem cells exhibited homogeneous spindle shape, and highly expressed CD44, CD73, CD90 and CD105, negatively expressed CD34, CD45, CD80 and CD86. After induced differentiation, the cells were positively stained by oil red O staining and alizarlin red staining, which were the markers of adipocytes and osteoblasts. The results of ELISA indicated that the expression level of interleukin-6 and interleukin-8 in the CD200~+ placenta-derived mesenchymal stem cells was strikingly higher than placenta-derived mesenchymal stem cells and CD200~-placenta-derived mesenchymal stem cells. Compared with normal and CD200~-placenta-derived mesenchymal stem cells, CD200~+ placenta-derived mesenchymal stem cells inhibited T cell proliferation significantly, lowered the levels of interferon-γ and interleukin-2, and upregulated the level of interleukin-10. We found that CD200~+ placenta-derived mesenchymal stem cells have better immunoregulatory capability compared with normal and CD200~-placenta-derived mesenchymal stem cells, which may provide an ideal theoretical basis for clinical use.
引文
[1]Schuleri KH, Boyle AJ, Hare JM.Mesenchymal stem cells for cardiac regenerative therapy.Handb Exp Pharmacol. 2007;(180):195-218.
[2]Deda H, Inci MC, Kürek?i AE, et al. Treatment of amyotrophic lateral sclerosis patients by autologous bone marrow-derived hematopoietic stem cell transplantation:a 1-year follow-up.Cytotherapy. 2009;11(1):18-25.
[3]Martinez HR, Gonzalez-Garza MT, Moreno-Cuevas JE, et al.Stem-cell transplantation into the frontal motor cortex in amyotrophic lateral sclerosis patients.Cytotherapy. 2009;11(1):26-34..
[4]Taupin P.OTI-010 Osiris Therapeutics/JCR Pharmaceuticals.Curr OpinInvestig Drugs. 2006;7(5):473-481.
[5]Abdi R, Fiorina P, Adra CN, et al. Immunomodulation by mesenchymal stem cells:a potential therapeutic strategy for type 1 diabetes.Diabetes. 2008;57(7):1759-1767.
[6]Yoshikawa T, Mitsuno H, Nonaka I, et al. Wound therapy by marrow mesenchymal cell transplantation.Plast Reconstr Surg. 2008;121(3):860-877.
[7]Le Blanc K, Tammik L, Sundberg B, et al. Mesenchymal stem cells inhibit and stimulate mixed lymphocyte cultures and mitogenic responses independently of the major histocompatibility complex.Scand J Immunol. 2003;57(1):11-20.
[8]Aggarwal S, Pittenger MF.Human mesenchymal stem cells modulate allogeneic immune cell responses.Blood. 2005;105(4):1815-1822.
[9]Jiang XX, Zhang Y, Liu B, et al. Human mesenchymal stem cells inhibit differentiation and function of monocyte-derived dendritic cells.Blood. 2005;105(10):4120-4126.
[10]Beyth S, Borovsky Z, Mevorach D, et al. Human mesenchymal stem cells alter antigen-presenting cell maturation and induce T-cell unresponsiveness.Blood.2005;105(5):2214-2219.
[11]Corcione A, Benvenuto F, Ferretti E, et al. Human mesenchymal stem cells modulate B-cell functions.Blood.2006;107(1):367-372.
[12]Chan JL, Tang KC, Patel AP, et al. Antigen-presenting property of mesenchymal stem cells occurs during a narrow window at low levels of interferon-gamma.Blood. 2006;107(12):4817-4824.
[13]Stagg J, Pommey S, Eliopoulos N, et al. Interferon-gammastimulated marrow stromal cells:a new type of nonhematopoietic antigen-presenting cell.Blood. 2006;107(6):2570-2577.
[14]Stagg J.Immune regulation by mesenchymal stem cells:two sides to the coin.Tissue Antigens. 2007;69(1):1-9.
[15]Nauta AJ, Westerhuis G, Kruisselbrink AB, et al.Donor-derived mesenchymal stem cells are immunogenic in an allogeneic host and stimulate donor graft rejection in a nonmyeloablative setting.Blood. 2006;108(6):2114-2120.
[16]郝贵亮,王立斌,陈冬梅,等.母体和胎儿来源的人胎盘间充质干细胞抑制小鼠皮肤移植免疫排斥作用的比较[J].细胞与分子免疫学杂志,2015,31(5):609-614.
[17]马晓娜,刘婷,金毅然,等.母体来源人胎盘间充质细胞的分离培养及其分化能力的研究[J].中国医药指南,2013,11(34):3-5.
[18]SykováE, JendelováP, UrdzíkováL, et al. Bone marrow stem cells and polymer hydrogels--two strategies for spinal cord injury repair.Cell Mol Neurobiol. 2006;26(7-8):1113-1129.
[19]Satija NK, Singh VK, Verma YK, et al. Mesenchymal stem cell-based therapy:a new paradigm in regenerative medicine.J Cell Mol Med. 2009;13(11-12):4385-4402.
[20]Patel DM, Shah J, Srivastava AS.Therapeutic potential of mesenchymal stem cells in regenerative medicine.Stem Cells Int. 2013;2013:496218.
[21]Parolini O, Alviano F, Bagnara GP, et al. Concise review:isolation and characterization of cells from human term placenta:outcome of the first international Workshop on Placenta Derived Stem Cells.Stem Cells. 2008;26(2):300-311.
[22]Kadam S, Muthyala S, Nair P, et al. Human placenta-derived mesenchymal stem cells and islet-like cell clusters generated from these cells as a novel source for stem cell therapy in diabetes.Rev Diabet Stud. 2010;7(2):168-182.
[23]Lu GH, Zhang SZ, Chen Q, et al. Isolation and multipotent differentiation of human decidua basalis-derived mesenchymal stem cells. Nan Fang Yi Ke Da Xue Xue Bao.2011;31(2):262-265.
[24]Miao Z, Jin J, Chen L, et al. Isolation of mesenchymal stem cells from human placenta:comparison with human bone marrow mesenchymal stem cells.Cell Biol Int. 2006;30(9):681-687.
[25]张红艳,杨乃龙.胎盘来源间充质干细胞的生物学特征[J].中国组织工程研究,2010,14(40):7535-7538.
[26]Alviano F, Fossati V, Marchionni C, et al. Term Amniotic membrane is a high throughput source for multipotent Mesenchymal Stem Cells with the ability to differentiate into endothelial cells in vitro.BMC Dev Biol. 2007;7:11.
[27]Wolbank S, Peterbauer A, Fahrner M, et al. Dose-dependent immunomodulatory effect of human stem cells from amniotic membrane:a comparison with human mesenchymal stem cells from adipose tissue.Tissue Eng. 2007;13(6):1173-1183.
[28]Portmann-Lanz CB, Schoeberlein A, Portmann R, et al.Turning placenta into brain:placental mesenchymal stem cells differentiate into neurons and oligodendrocytes.Am J Obstet Gynecol. 2010;202(3):294. e1-294. e11.
[29]Ventura C, Cantoni S, Bianchi F, et al. Hyaluronan mixed esters of butyric and retinoic Acid drive cardiac and endothelial fate in term placenta human mesenchymal stem cells and enhance cardiac repair in infarcted rat hearts.J Biol Chem. 2007;282(19):14243-14252.
[30]李治,赵伟,刘伟,等.玻璃酸钠及胎盘间充质干细胞和诱导的软骨细胞膝关节腔内注射:修复膝骨关节炎[J].中国组织工程研究,2014,18(50):8140-8146.
[31]Li Z, Qin H, Feng Z, et al. Human umbilical cord mesenchymal stem cell-loaded amniotic membrane for the repair of radial nerve injury.Neural Regen Res. 2013; 8(36):3441-3448.
[32]Ercal P, Pekozer GG, Gumru OZ, et al. Influence of STRO-1selection on osteogenic potential of human tooth germ derived mesenchymal stem cells.Arch Oral Biol. 2017;82:293-301.
[33]Najar M, Crompot E, van Grunsven LA, et al. Aldehyde Dehydrogenase Activity in Adipose Tissue:Isolation and Gene Expression Profile of Distinct Sub-population of Mesenchymal Stromal Cells.Stem Cell Rev. 2018; 14(4):599-611.
[34]Lee HT, Chang HT, Lee S, et al. Role of IGF1R(+)MSCs in modulating neuroplasticity via CXCR4 cross-interaction.Sci Rep. 2016;6:32595.
[35]Cao H, Yang J, Yu J, et al. Therapeutic potential of transplanted placental mesenchymal stem cells in treating Chinese miniature pigs with acute liver failure.BMC Med.2012;10:56.
[36]Geffner LF, Santacruz P, Izurieta M, et al. Administration of autologous bone marrow stem cells into spinal cord injury patients via multiple routes is safe and improves their quality of life:comprehensive case studies.Cell Transplant. 2008;17(12):1277-1293.
[37]Hoek RM, Ruuls SR, Murphy CA, et al. Down-regulation of the macrophage lineage through interaction with OX2(CD200).Science. 2000;290(5497):1768-1771.
[38]Jenmalm MC, Cherwinski H, Bowman EP, et al. Regulation of myeloid cell function through the CD200 receptor.J Immunol.2006;176(1):191-199.
[39]Gorczynski RM, Lee L, Boudakov I.Augmented Induction of CD4+CD25+Treg using monoclonal antibodies to CD200R.Transplantation. 2005;79(9):1180-1183.
[40]Kretz-Rommel A, Qin F, Dakappagari N, et al. CD200expression on tumor cells suppresses antitumor immunity:new approaches to cancer immunotherapy.J Immunol. 2007;178(9):5595-5605.
[41]Coles SJ, Hills RK, Wang EC, et al. Increased CD200expression in acute myeloid leukemia is linked with an increased frequency of FoxP3+regulatory T cells.Leukemia.2012;26(9):2146-2148.
[42]Guo Y, Xu F, Lu T, et al. Interleukin-6 signaling pathway in targeted therapy for cancer.Cancer Treat Rev. 2012; 38(7):904-910.
[43]Ha H, Debnath B, Neamati N.Role of the CXCL8-CXCR1/2Axis in Cancer and Inflammatory Diseases.Theranostics.2017;7(6):1543-1588.
[2]Deda H, Inci MC, Kürek?i AE, et al. Treatment of amyotrophic lateral sclerosis patients by autologous bone marrow-derived hematopoietic stem cell transplantation:a 1-year follow-up.Cytotherapy. 2009;11(1):18-25.
[3]Martinez HR, Gonzalez-Garza MT, Moreno-Cuevas JE, et al.Stem-cell transplantation into the frontal motor cortex in amyotrophic lateral sclerosis patients.Cytotherapy. 2009;11(1):26-34..
[4]Taupin P.OTI-010 Osiris Therapeutics/JCR Pharmaceuticals.Curr OpinInvestig Drugs. 2006;7(5):473-481.
[5]Abdi R, Fiorina P, Adra CN, et al. Immunomodulation by mesenchymal stem cells:a potential therapeutic strategy for type 1 diabetes.Diabetes. 2008;57(7):1759-1767.
[6]Yoshikawa T, Mitsuno H, Nonaka I, et al. Wound therapy by marrow mesenchymal cell transplantation.Plast Reconstr Surg. 2008;121(3):860-877.
[7]Le Blanc K, Tammik L, Sundberg B, et al. Mesenchymal stem cells inhibit and stimulate mixed lymphocyte cultures and mitogenic responses independently of the major histocompatibility complex.Scand J Immunol. 2003;57(1):11-20.
[8]Aggarwal S, Pittenger MF.Human mesenchymal stem cells modulate allogeneic immune cell responses.Blood. 2005;105(4):1815-1822.
[9]Jiang XX, Zhang Y, Liu B, et al. Human mesenchymal stem cells inhibit differentiation and function of monocyte-derived dendritic cells.Blood. 2005;105(10):4120-4126.
[10]Beyth S, Borovsky Z, Mevorach D, et al. Human mesenchymal stem cells alter antigen-presenting cell maturation and induce T-cell unresponsiveness.Blood.2005;105(5):2214-2219.
[11]Corcione A, Benvenuto F, Ferretti E, et al. Human mesenchymal stem cells modulate B-cell functions.Blood.2006;107(1):367-372.
[12]Chan JL, Tang KC, Patel AP, et al. Antigen-presenting property of mesenchymal stem cells occurs during a narrow window at low levels of interferon-gamma.Blood. 2006;107(12):4817-4824.
[13]Stagg J, Pommey S, Eliopoulos N, et al. Interferon-gammastimulated marrow stromal cells:a new type of nonhematopoietic antigen-presenting cell.Blood. 2006;107(6):2570-2577.
[14]Stagg J.Immune regulation by mesenchymal stem cells:two sides to the coin.Tissue Antigens. 2007;69(1):1-9.
[15]Nauta AJ, Westerhuis G, Kruisselbrink AB, et al.Donor-derived mesenchymal stem cells are immunogenic in an allogeneic host and stimulate donor graft rejection in a nonmyeloablative setting.Blood. 2006;108(6):2114-2120.
[16]郝贵亮,王立斌,陈冬梅,等.母体和胎儿来源的人胎盘间充质干细胞抑制小鼠皮肤移植免疫排斥作用的比较[J].细胞与分子免疫学杂志,2015,31(5):609-614.
[17]马晓娜,刘婷,金毅然,等.母体来源人胎盘间充质细胞的分离培养及其分化能力的研究[J].中国医药指南,2013,11(34):3-5.
[18]SykováE, JendelováP, UrdzíkováL, et al. Bone marrow stem cells and polymer hydrogels--two strategies for spinal cord injury repair.Cell Mol Neurobiol. 2006;26(7-8):1113-1129.
[19]Satija NK, Singh VK, Verma YK, et al. Mesenchymal stem cell-based therapy:a new paradigm in regenerative medicine.J Cell Mol Med. 2009;13(11-12):4385-4402.
[20]Patel DM, Shah J, Srivastava AS.Therapeutic potential of mesenchymal stem cells in regenerative medicine.Stem Cells Int. 2013;2013:496218.
[21]Parolini O, Alviano F, Bagnara GP, et al. Concise review:isolation and characterization of cells from human term placenta:outcome of the first international Workshop on Placenta Derived Stem Cells.Stem Cells. 2008;26(2):300-311.
[22]Kadam S, Muthyala S, Nair P, et al. Human placenta-derived mesenchymal stem cells and islet-like cell clusters generated from these cells as a novel source for stem cell therapy in diabetes.Rev Diabet Stud. 2010;7(2):168-182.
[23]Lu GH, Zhang SZ, Chen Q, et al. Isolation and multipotent differentiation of human decidua basalis-derived mesenchymal stem cells. Nan Fang Yi Ke Da Xue Xue Bao.2011;31(2):262-265.
[24]Miao Z, Jin J, Chen L, et al. Isolation of mesenchymal stem cells from human placenta:comparison with human bone marrow mesenchymal stem cells.Cell Biol Int. 2006;30(9):681-687.
[25]张红艳,杨乃龙.胎盘来源间充质干细胞的生物学特征[J].中国组织工程研究,2010,14(40):7535-7538.
[26]Alviano F, Fossati V, Marchionni C, et al. Term Amniotic membrane is a high throughput source for multipotent Mesenchymal Stem Cells with the ability to differentiate into endothelial cells in vitro.BMC Dev Biol. 2007;7:11.
[27]Wolbank S, Peterbauer A, Fahrner M, et al. Dose-dependent immunomodulatory effect of human stem cells from amniotic membrane:a comparison with human mesenchymal stem cells from adipose tissue.Tissue Eng. 2007;13(6):1173-1183.
[28]Portmann-Lanz CB, Schoeberlein A, Portmann R, et al.Turning placenta into brain:placental mesenchymal stem cells differentiate into neurons and oligodendrocytes.Am J Obstet Gynecol. 2010;202(3):294. e1-294. e11.
[29]Ventura C, Cantoni S, Bianchi F, et al. Hyaluronan mixed esters of butyric and retinoic Acid drive cardiac and endothelial fate in term placenta human mesenchymal stem cells and enhance cardiac repair in infarcted rat hearts.J Biol Chem. 2007;282(19):14243-14252.
[30]李治,赵伟,刘伟,等.玻璃酸钠及胎盘间充质干细胞和诱导的软骨细胞膝关节腔内注射:修复膝骨关节炎[J].中国组织工程研究,2014,18(50):8140-8146.
[31]Li Z, Qin H, Feng Z, et al. Human umbilical cord mesenchymal stem cell-loaded amniotic membrane for the repair of radial nerve injury.Neural Regen Res. 2013; 8(36):3441-3448.
[32]Ercal P, Pekozer GG, Gumru OZ, et al. Influence of STRO-1selection on osteogenic potential of human tooth germ derived mesenchymal stem cells.Arch Oral Biol. 2017;82:293-301.
[33]Najar M, Crompot E, van Grunsven LA, et al. Aldehyde Dehydrogenase Activity in Adipose Tissue:Isolation and Gene Expression Profile of Distinct Sub-population of Mesenchymal Stromal Cells.Stem Cell Rev. 2018; 14(4):599-611.
[34]Lee HT, Chang HT, Lee S, et al. Role of IGF1R(+)MSCs in modulating neuroplasticity via CXCR4 cross-interaction.Sci Rep. 2016;6:32595.
[35]Cao H, Yang J, Yu J, et al. Therapeutic potential of transplanted placental mesenchymal stem cells in treating Chinese miniature pigs with acute liver failure.BMC Med.2012;10:56.
[36]Geffner LF, Santacruz P, Izurieta M, et al. Administration of autologous bone marrow stem cells into spinal cord injury patients via multiple routes is safe and improves their quality of life:comprehensive case studies.Cell Transplant. 2008;17(12):1277-1293.
[37]Hoek RM, Ruuls SR, Murphy CA, et al. Down-regulation of the macrophage lineage through interaction with OX2(CD200).Science. 2000;290(5497):1768-1771.
[38]Jenmalm MC, Cherwinski H, Bowman EP, et al. Regulation of myeloid cell function through the CD200 receptor.J Immunol.2006;176(1):191-199.
[39]Gorczynski RM, Lee L, Boudakov I.Augmented Induction of CD4+CD25+Treg using monoclonal antibodies to CD200R.Transplantation. 2005;79(9):1180-1183.
[40]Kretz-Rommel A, Qin F, Dakappagari N, et al. CD200expression on tumor cells suppresses antitumor immunity:new approaches to cancer immunotherapy.J Immunol. 2007;178(9):5595-5605.
[41]Coles SJ, Hills RK, Wang EC, et al. Increased CD200expression in acute myeloid leukemia is linked with an increased frequency of FoxP3+regulatory T cells.Leukemia.2012;26(9):2146-2148.
[42]Guo Y, Xu F, Lu T, et al. Interleukin-6 signaling pathway in targeted therapy for cancer.Cancer Treat Rev. 2012; 38(7):904-910.
[43]Ha H, Debnath B, Neamati N.Role of the CXCL8-CXCR1/2Axis in Cancer and Inflammatory Diseases.Theranostics.2017;7(6):1543-1588.