慢病毒载体介导人CD1d基因转染PANC-1实验研究
|
摘要
目的为了寻找胰腺癌基因治疗新方法,利用基因克隆、慢病毒载体构建、细胞转染等方法使人CDld基因稳定转染胰腺癌细胞株PANC-1,为后期以CDld-NKT为桥梁,研究细胞免疫原性变化、免疫上下游通路奠定基础。
方法利用基因克隆技术克隆出人CDld基因,连接在pEASY-Tl simple cloning vector上,构建成pEASY-Tl simple-CDld质粒,再进行序列测定；利用基因重组技术将人CDld基因连接慢病毒载体PCDH-CMV-MCS-EFl-copGFP,进行病毒包装、滴度测定后浓缩纯化；通过慢病毒转染预实验确定MOI,进行人CDld重组慢病毒载体转染胰腺癌PANC-1细胞株。
结果克隆出人CD1d基因,成功构建pEASY-Tl simple-CDld质粒,测序结果显示与人类基因库中CD1d基因序列一致；成功构建慢病毒载体质粒PCDH-CMV-MCS-EFl-copGFP-CDld,经病毒包装后得到携带CD1d的重组慢病毒颗粒Lenti-hCDld,浓缩后病毒滴度达到4.4×108TU/ml；重组慢病毒颗粒Lenti-hCDld成功转染胰腺癌细胞株PANC-1。
结论可以通过基因克隆、慢病毒载体构建、细胞转染等方法稳定转染CD1d基因至胰腺癌细胞株PANC-1,这为后期以CDld-NKT为桥梁,进行细胞免疫原性变化、免疫上下游通路的研究和胰腺癌的基因治疗奠定了基础。
Objective Use of gene clone, lentiviral vector construction, cell transfection methods to make people CDld gene stability transfection pancreatic cancer cell lines PANC-1, provide a new perspective to explore the pancreatic cancer gene therapy, lay the foundation of studying the cells immunogenicity change, upstream and downstream immune pathways using the CDld-NKT as the bridge.
Methods Human CDld gene was cloned by cloning technology,which connected to the pEASY-Tl simple cloning vector,and then sequenced;By recombinant technology.human CDld gene was enclosed to lentiviral vector PCDH-CMV-MCS-EFl-copGFP,which was concentrated and purificated after determining the concentration;MOI was determined by pre-test of lentiviral transfection.PANC-1cell line was transfected by lentivirus-mediated hCDld.
Results Human CDld gene was successfully cloned and constructed into pEASY-Tl-simple-CDld plasmid.It was showed that the cloned hCDld gene is identical with the human genebank by sequencry;Lentivirus vector(PCDH-CMV-MCS-EFl-copGFP-CDld)was successfullyconstructed,and the mature Lenti-hCDld was packaged,and the virus titer of which concentrated to4.4xlO8TU/ml;Lenti-hCDld stably transfected pancreatic cancer cell line PANC-1.
Conclusion Human CDld gene can be transfected by gene clone, lentiviral vector construction, cell transfection methods to pancreatic cancer cell lines PANC-1, lay the foundation of studying the cells immunogenicity change, upstream and downstream immune pathways using the CDld-NKT as the bridge, provide a new perspective to explore the pancreatic cancer gene therapy.
方法利用基因克隆技术克隆出人CDld基因,连接在pEASY-Tl simple cloning vector上,构建成pEASY-Tl simple-CDld质粒,再进行序列测定；利用基因重组技术将人CDld基因连接慢病毒载体PCDH-CMV-MCS-EFl-copGFP,进行病毒包装、滴度测定后浓缩纯化；通过慢病毒转染预实验确定MOI,进行人CDld重组慢病毒载体转染胰腺癌PANC-1细胞株。
结果克隆出人CD1d基因,成功构建pEASY-Tl simple-CDld质粒,测序结果显示与人类基因库中CD1d基因序列一致；成功构建慢病毒载体质粒PCDH-CMV-MCS-EFl-copGFP-CDld,经病毒包装后得到携带CD1d的重组慢病毒颗粒Lenti-hCDld,浓缩后病毒滴度达到4.4×108TU/ml；重组慢病毒颗粒Lenti-hCDld成功转染胰腺癌细胞株PANC-1。
结论可以通过基因克隆、慢病毒载体构建、细胞转染等方法稳定转染CD1d基因至胰腺癌细胞株PANC-1,这为后期以CDld-NKT为桥梁,进行细胞免疫原性变化、免疫上下游通路的研究和胰腺癌的基因治疗奠定了基础。
Objective Use of gene clone, lentiviral vector construction, cell transfection methods to make people CDld gene stability transfection pancreatic cancer cell lines PANC-1, provide a new perspective to explore the pancreatic cancer gene therapy, lay the foundation of studying the cells immunogenicity change, upstream and downstream immune pathways using the CDld-NKT as the bridge.
Methods Human CDld gene was cloned by cloning technology,which connected to the pEASY-Tl simple cloning vector,and then sequenced;By recombinant technology.human CDld gene was enclosed to lentiviral vector PCDH-CMV-MCS-EFl-copGFP,which was concentrated and purificated after determining the concentration;MOI was determined by pre-test of lentiviral transfection.PANC-1cell line was transfected by lentivirus-mediated hCDld.
Results Human CDld gene was successfully cloned and constructed into pEASY-Tl-simple-CDld plasmid.It was showed that the cloned hCDld gene is identical with the human genebank by sequencry;Lentivirus vector(PCDH-CMV-MCS-EFl-copGFP-CDld)was successfullyconstructed,and the mature Lenti-hCDld was packaged,and the virus titer of which concentrated to4.4xlO8TU/ml;Lenti-hCDld stably transfected pancreatic cancer cell line PANC-1.
Conclusion Human CDld gene can be transfected by gene clone, lentiviral vector construction, cell transfection methods to pancreatic cancer cell lines PANC-1, lay the foundation of studying the cells immunogenicity change, upstream and downstream immune pathways using the CDld-NKT as the bridge, provide a new perspective to explore the pancreatic cancer gene therapy.
引文
[1]Koch M, Stronge VS, Shepherd D, et al. The crystal structure of human CDld with and witho- ut a-galactosylceramide [J]. Nat Immunol, 2005,6 (8):819-826.
[2]Lee A, Farrand KJ, Dickgreber N, et al. Novel synthesis of [alpha]-galactosyl-ceramides and c-onfirmation of their powerful NKT cell agonist activity [J]. Carbohydr Res, 2006, 341 (17):2785-2798.
[3]Ragin MJ, Sahu N, August A. Differential regulation of cytokine production by CD ld-restricted NKT cells in response to superantigen staphylococcal enterotoxin B exposure [J]. Infect Immun, 2006,74(1):282-288.
[4]Saito S, Sakai M, Sasaki Y, et al. Quantitative analysis of peripheral blood ThO, Thl, Th2 and the Thl:Th2 cell ratio during normal human pregnancy and preeclampsia[J]. Clinical and experi-mental immunology, 1999,117 (3):550-555.
[5]Morrow JF,Berg P.Cleavage of Simian virus 40 DNA at a unique site by a bacterial restriction enzyme[J]. Proc Natl Acad Sci USA.l972,69 (11):3365-9.
[6]Hassanin H, Serba S, Schmidt J, et al. Ex vivo expanded telomerase-specific T cells are effective in an orthotopic mouse model for pancreatic adenocarcinoma [J]. Clin Exp Immunol, 2009, 158(1): 125-132.
[7]Schrantz N, Sagiv Y, Liu Y, et al . The Niemann-Pick type C2 protein loads isoglobotrihexos-ylceramide onto CDld molecules and contributes to the thymic selection of NKT cells [J] . J Exp Med,2007,204 (7):841-852.
[8]陆田田,黄震,陈章权.CDld分子的结构与功能[J].生命的化学,2008,28(2)159-161.
[9]Zajonc DM, Elsliger MA, Teyton L, et al. Crystal structure of CDla in complex with a sulfatide self antigen at a resolution of 2.15A [J]. Nat Immunol, 2003,4 (8):808-815.
[10]Lawton AP, Prigozy TI, Brossay L, et al. The mouse CDld cytoplasmic tail mediates CDld trafficking and antigen presentation by adaptor protein 3-dependent and-independent mechanisms [J]. Immunol. 2005,174 (6):3179-3186.
[11]Kai C, Wenhua Z, Changliang W, et al. The CXCR4-CXCL12 pathway facilitates the progression of pancreatic cancer via Induction of angiogenesis and lymphangiogenesis [J]. Journal of Surgical Research,2010, 171(11): 143-150.
[12]Zhang D, Ma Q, Shen S, et al. Inhibition of pancreatic cancer cell proliferation by propranolol occurs through apoptosis Induction [J]. Pancreas.2009,38(1):94-100.
[13]Thomas RM, Ahmad SA. Current concepts in the surgical management of pancreatic cancer[J]. Surg Oncol Clin N Am.2010, 19(2):335-358.
[1]Delenda C.Lentiviral vectors:optimization of packaging.transduction and gene expression[J]. JGene Med,2004.6(Suppl1):S125-S138.
[2]Lois C,Refaeli Y,Qin XF,et al.Retroviruses as tools to study the immune system[J].Curr Opin Immunl,2001,13(4):496-504.
[3]Wong LF,Goodhead L.Prat C,et al.Lentivirus-mediated gene transfer to the central nervous system:therapeutic and research applications[J].Hum Gene Ther,2006,17(1):1-9.
[4]Dull T, Zuferey R, Kelly M, et al. A third-generation lentivirus vector with a conditional packaging system[J]. Journal of virology.1998,72(11):8463-8471
[5]Naldini L, Blomer U, Gallay P, et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector[J]. Science.1996,272:263-267
[6]Dougherty J P, Temin H M. A promoterless retroviral vector indicates that there are sequences in U3 required for 3'RNA processing[J]. Proceeding of the Natoinal Academy of Sciences of the USA.1987,84(5):1197-1201
[7]Yu SF, von Ruden T, Kantof PW, et al. Self-inactivating retroviral vectors designed for transfer of whole genes into mammalian cells[J]. Proceeding of the Natoinal Academy of Sciences of the USA. 1986,83(10):3194-3198
[8]Schraufstatter IU,Trieu K.Zhao M.et al.IL-8-mediated cell migration in endothelial cells depends on cathepsin B activity and transactivation of the epidermal growth factor receptor[J].J Immunol,2003,171(12):6714-6722.
[9]Heidemann J,Ogawa H.Dwinell MB,et al.Angiogenic effects of interleukin 8(CXCL8)in human intestinal microvascular endothelial cells are mediated by CXCR2[J].J Biol Chem,2003,278(10):8508-8515.
[10]Keane MP,Belperio JA,Xue YY,et al.Depletion of CXCR2 inhibits tumor growth and angiogenesis in a murine model of lung cancer[J].J Immunol,2004,172(5):2853-6280.
[11]Magda Kucia,Ryan Reca.Katarzyna Miekus,et al.Trafficking of Normal Stem Cells and Metastasis of Cancer Stem Cells Involve Similar Mechanisms:Pivotal Role of the SDF-1-CXCR4 Axis[J].Stem Cells,2005,23(7):8791-8794.
[12]Albert Zlotnik.Chemokines in neoplastic progression[J].Semin Cancer Biol,2004,14(3):181-185.
[13]Phillips RJ,Mestas J,Gharaee-Kermani M,et al.Epidermal growth factor and hypoxia-induced expression of CXC chemokine receptor 4 on non-small cell lung cancer cells is regulated by the phosphatidylinositol 3-kinase/PTEN/AKT/mammalian target of rapamycin signaling pathway andactivation of hypoxia inducible factor-1 alpha[J].J Biol Chem,2005,280(23):22473-22481.
[14]Joshi SK, Lang GA, Devera TS, et al. Differential contribution of dendritic cell CDld to NKT cell-enhanced humoral immunity and CD8+T cell activation[J]. Leukoc Biol 2012,91 (2):782-785.
[1]Koch M, Stronge VS, Shepherd D, et al. The crystal structure of human CD ld with and witho- ut a-galactosylceramide [J]. Nat Immunol, 2005,6 (8):819-826.
[2]Schrantz N, Sagiv Y, Liu Y, et al . The Niemann-Pick type C2 protein loads isoglobotrihexos-ylceramide onto CDld molecules and contributes to the thymic selection of NKT cells [J]. J Exp Med,2007,204 (7):841-852.
[3]陆田田,黄震,陈章权.CD1d分子的结构与功能[J].生命的化学,2008,28(2)159-161.
[4]Zajonc DM, Elsliger MA, Teyton L, et al. Crystal structure of CDla in complex with a sulfatide self antigen at a resolution of 2.15A [J]. Nat Immunol,2003,4 (8):808-815.
[5]Lawton AP, Prigozy TI, Brossay L, et al. The mouse CDld cytoplasmic tail mediates CDld tr-afficking and antigen presentation by adaptor protein 3-dependent and-independent mechanisms [J]. Immunol,2005,174 (6):3179-3186.
[6]Kang SJ, Cresswell P. Regulation of intracellular trafficking of human CD Id by association w- ith MHC class II molecules [J]. EMBO,2002,21 (7):1650-1660.
[7]Honey K, Benlagha K, Beers C, et al. Thymocyte expression of cathepsin L is essential for N- KT cell development [J]. Nat Immunol,2002,3(11):1069-1074.
[8]Thierry M, Andrew J, James P, et al. A molecular basis for NKT cell recognition of CDld-self--antigen [J]. immunity, 2011,3 (5):315-326.
[9]Sharif S, Arreaza GA, Zucker P, et al. Activation of natural killer T cells by alpha-galactosyl-ceramide treatment prevents the onset and recurrence of autoimmune Type 1 diabetes[J]. Nat Med, 2010,7(5):1057-1062.
[10]Lee A, Farrand KJ, Dickgreber N, et al. Novel synthesis of [alpha]-galactosyl-ceramides and c-onfirmation of their powerful NKT cell agonist activity [J] . Carbohydr Res, 2006, 341 (17):2785-2798.
[11]Prigozy TI, Naidenko O, Qasba P, et al. Glycolipid antigen processing for presentation by CD- 1d molecules [J] . Science, 2001,291 (5504):664-667.
[12]Dougan SK, Salas A, Rava P, et al. Microsomal triglyceride transfer protein lipidation and co-ntrol of CDld on antigen-presenting cells [J]. Exp Med, 2005,202 (4):529-539.
[13]刘景华,窦立萍,王莉莉等.CDld四聚体检测NKT细胞[J].细胞与分子免疫学杂志2009,25(1)：82-83.
[14]Ragin MJ, Sahu N, August A. Differential regulation of cytokine production by CD Id-restricted NKT cells in response to superantigen staphylococcal enterotoxin B exposure [J]. Infect Immun, 2006,74(1):282-288.
[15]Saito S, Sakai M, Sasaki Y, et al. Quantitative analysis of peripheral blood ThO, Thl, Th2 and the Thl:Th2 cell ratio during normal human pregnancy and preeclampsia[J] . Clinical and experi-mental immunology, 1999, 117 (3):550-555.
[16]Matsuda J L, Naidenko O V, Gapin L, et al . Tracking the response of natural kill-er T cells to a glycolipid antigen using CDld tetramers[J]. J Exp Med,2000 19 (2):7-41-754.
[17]Bendelac A, Savage PB, Teyton L. The biology of NKT cells [J]. Annu Rev Immunol,2007,25 (4):297-336.
[18]Ishikawa A, Motohashi S, Ishikawa E, et al. A phase I study of alpha-Galactosylceramide (KRN7000)-pulsed dendritic cells in patients with advanced and recurrent non-small cell lung ca-ncer [J]. Clin Cancer Res,2005,11 (5):1910 -1917.
[19]Chang D, Osman K, Connolly J, et al. Sustained expansion of NKT cells and antigen-specific T cells after injection of alpha-galactosylceramide loaded mature dendritic cells in cancer patients [J]. Exp Med, 2005,201 (9):1503 - 1517.
[20]Raghuraman G, Geng Y, Wang CR. IFN-b-Mediated Up-Regulation of CDld in Bacteria-Inf-ected APCs [J]. J Immunol,2006,177 (11):7841-7848.
[21]Kojo S, Tsutsumi A, Goto D, et al. Low expression levels of soluble CDld gene in patients w with rheumatoid arthritis [J]. J Rheumatol, 2003,30 (12):2524-2528.
[22]Yang JQ, Chun T, Liu HZ, et al. CDld deficiency exacerbates inflammatory dermatitis in M- RL-lpr/lpr mice [J]. Eur J Immunol, 2004, 34 (6):1723-1732.
[23]Chen N, McCarthy C, Drakesmith H, et al. HIV-1 down-regulates the expression of CDld via Nef [J]. Eur J Immunol, 2006,36 (2):278-286.
[2]Lee A, Farrand KJ, Dickgreber N, et al. Novel synthesis of [alpha]-galactosyl-ceramides and c-onfirmation of their powerful NKT cell agonist activity [J]. Carbohydr Res, 2006, 341 (17):2785-2798.
[3]Ragin MJ, Sahu N, August A. Differential regulation of cytokine production by CD ld-restricted NKT cells in response to superantigen staphylococcal enterotoxin B exposure [J]. Infect Immun, 2006,74(1):282-288.
[4]Saito S, Sakai M, Sasaki Y, et al. Quantitative analysis of peripheral blood ThO, Thl, Th2 and the Thl:Th2 cell ratio during normal human pregnancy and preeclampsia[J]. Clinical and experi-mental immunology, 1999,117 (3):550-555.
[5]Morrow JF,Berg P.Cleavage of Simian virus 40 DNA at a unique site by a bacterial restriction enzyme[J]. Proc Natl Acad Sci USA.l972,69 (11):3365-9.
[6]Hassanin H, Serba S, Schmidt J, et al. Ex vivo expanded telomerase-specific T cells are effective in an orthotopic mouse model for pancreatic adenocarcinoma [J]. Clin Exp Immunol, 2009, 158(1): 125-132.
[7]Schrantz N, Sagiv Y, Liu Y, et al . The Niemann-Pick type C2 protein loads isoglobotrihexos-ylceramide onto CDld molecules and contributes to the thymic selection of NKT cells [J] . J Exp Med,2007,204 (7):841-852.
[8]陆田田,黄震,陈章权.CDld分子的结构与功能[J].生命的化学,2008,28(2)159-161.
[9]Zajonc DM, Elsliger MA, Teyton L, et al. Crystal structure of CDla in complex with a sulfatide self antigen at a resolution of 2.15A [J]. Nat Immunol, 2003,4 (8):808-815.
[10]Lawton AP, Prigozy TI, Brossay L, et al. The mouse CDld cytoplasmic tail mediates CDld trafficking and antigen presentation by adaptor protein 3-dependent and-independent mechanisms [J]. Immunol. 2005,174 (6):3179-3186.
[11]Kai C, Wenhua Z, Changliang W, et al. The CXCR4-CXCL12 pathway facilitates the progression of pancreatic cancer via Induction of angiogenesis and lymphangiogenesis [J]. Journal of Surgical Research,2010, 171(11): 143-150.
[12]Zhang D, Ma Q, Shen S, et al. Inhibition of pancreatic cancer cell proliferation by propranolol occurs through apoptosis Induction [J]. Pancreas.2009,38(1):94-100.
[13]Thomas RM, Ahmad SA. Current concepts in the surgical management of pancreatic cancer[J]. Surg Oncol Clin N Am.2010, 19(2):335-358.
[1]Delenda C.Lentiviral vectors:optimization of packaging.transduction and gene expression[J]. JGene Med,2004.6(Suppl1):S125-S138.
[2]Lois C,Refaeli Y,Qin XF,et al.Retroviruses as tools to study the immune system[J].Curr Opin Immunl,2001,13(4):496-504.
[3]Wong LF,Goodhead L.Prat C,et al.Lentivirus-mediated gene transfer to the central nervous system:therapeutic and research applications[J].Hum Gene Ther,2006,17(1):1-9.
[4]Dull T, Zuferey R, Kelly M, et al. A third-generation lentivirus vector with a conditional packaging system[J]. Journal of virology.1998,72(11):8463-8471
[5]Naldini L, Blomer U, Gallay P, et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector[J]. Science.1996,272:263-267
[6]Dougherty J P, Temin H M. A promoterless retroviral vector indicates that there are sequences in U3 required for 3'RNA processing[J]. Proceeding of the Natoinal Academy of Sciences of the USA.1987,84(5):1197-1201
[7]Yu SF, von Ruden T, Kantof PW, et al. Self-inactivating retroviral vectors designed for transfer of whole genes into mammalian cells[J]. Proceeding of the Natoinal Academy of Sciences of the USA. 1986,83(10):3194-3198
[8]Schraufstatter IU,Trieu K.Zhao M.et al.IL-8-mediated cell migration in endothelial cells depends on cathepsin B activity and transactivation of the epidermal growth factor receptor[J].J Immunol,2003,171(12):6714-6722.
[9]Heidemann J,Ogawa H.Dwinell MB,et al.Angiogenic effects of interleukin 8(CXCL8)in human intestinal microvascular endothelial cells are mediated by CXCR2[J].J Biol Chem,2003,278(10):8508-8515.
[10]Keane MP,Belperio JA,Xue YY,et al.Depletion of CXCR2 inhibits tumor growth and angiogenesis in a murine model of lung cancer[J].J Immunol,2004,172(5):2853-6280.
[11]Magda Kucia,Ryan Reca.Katarzyna Miekus,et al.Trafficking of Normal Stem Cells and Metastasis of Cancer Stem Cells Involve Similar Mechanisms:Pivotal Role of the SDF-1-CXCR4 Axis[J].Stem Cells,2005,23(7):8791-8794.
[12]Albert Zlotnik.Chemokines in neoplastic progression[J].Semin Cancer Biol,2004,14(3):181-185.
[13]Phillips RJ,Mestas J,Gharaee-Kermani M,et al.Epidermal growth factor and hypoxia-induced expression of CXC chemokine receptor 4 on non-small cell lung cancer cells is regulated by the phosphatidylinositol 3-kinase/PTEN/AKT/mammalian target of rapamycin signaling pathway andactivation of hypoxia inducible factor-1 alpha[J].J Biol Chem,2005,280(23):22473-22481.
[14]Joshi SK, Lang GA, Devera TS, et al. Differential contribution of dendritic cell CDld to NKT cell-enhanced humoral immunity and CD8+T cell activation[J]. Leukoc Biol 2012,91 (2):782-785.
[1]Koch M, Stronge VS, Shepherd D, et al. The crystal structure of human CD ld with and witho- ut a-galactosylceramide [J]. Nat Immunol, 2005,6 (8):819-826.
[2]Schrantz N, Sagiv Y, Liu Y, et al . The Niemann-Pick type C2 protein loads isoglobotrihexos-ylceramide onto CDld molecules and contributes to the thymic selection of NKT cells [J]. J Exp Med,2007,204 (7):841-852.
[3]陆田田,黄震,陈章权.CD1d分子的结构与功能[J].生命的化学,2008,28(2)159-161.
[4]Zajonc DM, Elsliger MA, Teyton L, et al. Crystal structure of CDla in complex with a sulfatide self antigen at a resolution of 2.15A [J]. Nat Immunol,2003,4 (8):808-815.
[5]Lawton AP, Prigozy TI, Brossay L, et al. The mouse CDld cytoplasmic tail mediates CDld tr-afficking and antigen presentation by adaptor protein 3-dependent and-independent mechanisms [J]. Immunol,2005,174 (6):3179-3186.
[6]Kang SJ, Cresswell P. Regulation of intracellular trafficking of human CD Id by association w- ith MHC class II molecules [J]. EMBO,2002,21 (7):1650-1660.
[7]Honey K, Benlagha K, Beers C, et al. Thymocyte expression of cathepsin L is essential for N- KT cell development [J]. Nat Immunol,2002,3(11):1069-1074.
[8]Thierry M, Andrew J, James P, et al. A molecular basis for NKT cell recognition of CDld-self--antigen [J]. immunity, 2011,3 (5):315-326.
[9]Sharif S, Arreaza GA, Zucker P, et al. Activation of natural killer T cells by alpha-galactosyl-ceramide treatment prevents the onset and recurrence of autoimmune Type 1 diabetes[J]. Nat Med, 2010,7(5):1057-1062.
[10]Lee A, Farrand KJ, Dickgreber N, et al. Novel synthesis of [alpha]-galactosyl-ceramides and c-onfirmation of their powerful NKT cell agonist activity [J] . Carbohydr Res, 2006, 341 (17):2785-2798.
[11]Prigozy TI, Naidenko O, Qasba P, et al. Glycolipid antigen processing for presentation by CD- 1d molecules [J] . Science, 2001,291 (5504):664-667.
[12]Dougan SK, Salas A, Rava P, et al. Microsomal triglyceride transfer protein lipidation and co-ntrol of CDld on antigen-presenting cells [J]. Exp Med, 2005,202 (4):529-539.
[13]刘景华,窦立萍,王莉莉等.CDld四聚体检测NKT细胞[J].细胞与分子免疫学杂志2009,25(1)：82-83.
[14]Ragin MJ, Sahu N, August A. Differential regulation of cytokine production by CD Id-restricted NKT cells in response to superantigen staphylococcal enterotoxin B exposure [J]. Infect Immun, 2006,74(1):282-288.
[15]Saito S, Sakai M, Sasaki Y, et al. Quantitative analysis of peripheral blood ThO, Thl, Th2 and the Thl:Th2 cell ratio during normal human pregnancy and preeclampsia[J] . Clinical and experi-mental immunology, 1999, 117 (3):550-555.
[16]Matsuda J L, Naidenko O V, Gapin L, et al . Tracking the response of natural kill-er T cells to a glycolipid antigen using CDld tetramers[J]. J Exp Med,2000 19 (2):7-41-754.
[17]Bendelac A, Savage PB, Teyton L. The biology of NKT cells [J]. Annu Rev Immunol,2007,25 (4):297-336.
[18]Ishikawa A, Motohashi S, Ishikawa E, et al. A phase I study of alpha-Galactosylceramide (KRN7000)-pulsed dendritic cells in patients with advanced and recurrent non-small cell lung ca-ncer [J]. Clin Cancer Res,2005,11 (5):1910 -1917.
[19]Chang D, Osman K, Connolly J, et al. Sustained expansion of NKT cells and antigen-specific T cells after injection of alpha-galactosylceramide loaded mature dendritic cells in cancer patients [J]. Exp Med, 2005,201 (9):1503 - 1517.
[20]Raghuraman G, Geng Y, Wang CR. IFN-b-Mediated Up-Regulation of CDld in Bacteria-Inf-ected APCs [J]. J Immunol,2006,177 (11):7841-7848.
[21]Kojo S, Tsutsumi A, Goto D, et al. Low expression levels of soluble CDld gene in patients w with rheumatoid arthritis [J]. J Rheumatol, 2003,30 (12):2524-2528.
[22]Yang JQ, Chun T, Liu HZ, et al. CDld deficiency exacerbates inflammatory dermatitis in M- RL-lpr/lpr mice [J]. Eur J Immunol, 2004, 34 (6):1723-1732.
[23]Chen N, McCarthy C, Drakesmith H, et al. HIV-1 down-regulates the expression of CDld via Nef [J]. Eur J Immunol, 2006,36 (2):278-286.