桿狀病毒在基因傳遞應用於間葉幹細胞上為一新穎且具有遠景的載體，且已應用於骨組織工程上，然而桿狀病毒轉導間葉幹細胞後是否會改變其免疫特性卻尚不明瞭，針對此問題，我們利用即時偵測同步定量反轉錄聚合□連鎖反應(qRT-PCR)，探討間葉幹細胞經由桿狀病毒轉導後是否會引起發炎相關細胞激素的表現或分泌。我們發現桿狀病毒轉導間葉幹細胞會誘發IL-1β, IL-6, IFN-□這三種細胞激素的基因表現，但不會誘發TNF-□與IFN-□。但以酵素免疫分析法(ELISA)定量時，我們發現IL-1β與IFN-□被桿狀病毒誘發的蛋白質表現量低於ELISA套組偵測濃度，只有IL-6被誘發的表現量可被偵測到，其分泌量隨時間增加逐漸降低，並於三日後降至與未轉導細胞相同。我們以螢光抗體標定間葉幹細胞表面上主要組織相容性復合體major histocompatibility complex (人類細胞上稱為Human Leukocyte Antigen, HLA)，再配合流式細胞儀分析，也發現桿狀病毒轉導幹細胞會在初期輕微影響其免疫特性(HLA-I增量表現)，但不影響HLA-II的表現特徵。間葉幹細胞和淋巴球細胞共同培養的實驗則顯示桿狀病毒不影響間葉幹細胞抑制淋巴球細胞增生的特性。在體外實驗的評估之後，我們將轉導後的間葉幹細胞植入大鼠腿部四頭肌，藉由此動物模式探討活體內的實驗。在H&E染色以及NUMA染色結果中，我們發現在第1, 2週時植入部位有大量的免疫細胞侵入，且幹細胞在第三週的存活率也很低。螢光免疫染色結果也顯示巨噬細胞、CD8+ T cells在第1, 2週時大量侵入植入部位。而在大鼠血液中巨噬細胞的變化，以免疫螢光抗體標定後由流式細胞儀定量分析也有同樣的趨勢。以上所有體外及體內實驗皆以轉導後之間葉幹細胞當成實驗組，假性轉導當成對照組，在活體內實驗中實驗組和對照組皆引發嚴重的免疫反應，因此推測可能是因異種移植所造成，而不是桿狀病毒轉導之影響。本研究顯示桿狀病毒不會引起間葉幹細胞多餘的體外抗病毒反應及影響體內的免疫反應，這有助於未來在各方面的應用。

Abe T, Takahashi H, Hamazaki H, Miyano-Kurosaki N, Matsuura Y, Takaku H. 2003. Baculovirus induces an innate immune response and confers protection from lethal influenza virus infection in mice. J Immunol 171(3):1133-9.
Airenne KJ, Hiltunen MO, Turunen MP, Turunen AM, Laitinen OH, Kulomaa MS, Yla-Herttuala S. 2000. Baculovirus-mediated periadventitial gene transfer to rabbit carotid artery. Gene Ther 7(17):1499-504.
Beck NB, Sidhu JS, Omiecinski CJ. 2000. Baculovirus vectors repress phenobarbital-mediated gene induction and stimulate cytokine expression in primary cultures of rat hepatocytes. Gene Ther 7(15):1274-83.
Bilello JP, Cable EE, Myers RL, Isom HC. 2003. Role of paracellular junction complexes in baculovirus-mediated gene transfer to nondividing rat hepatocytes. Gene Ther 10(9):733-49.
Bruder SP, Kurth AA, Shea M, Hayes WC, Jaiswal N, Kadiyala S. 1998. Bone regeneration by implantation of purified, culture-expanded human mesenchymal stem cells. J Orthop Res 16(2):155-62.
Caplan AI, Bruder SP. 2001. Mesenchymal stem cells: building blocks for molecular medicine in the 21st century. Trends Mol Med 7(6):259-64.
Check E. 2002. Gene therapy: a tragic setback. Nature 420:116-118.
Chen D, Murphy B, Sung R, Bromberg JS. 2003. Adaptive and innate immune responses to gene transfer vectors: role of cytokines and chemokines in vector function. Gene Ther 10(11):991-8.
Chen X, Armstrong MA, Li G. 2006. Mesenchymal stem cells in immunoregulation. Immunol Cell Biol 84(5):413-21.
Cizkova D, Rosocha J, Vanicky I, Jergova S, Cizek M. 2006. Transplants of human mesenchymal stem cells improve functional recovery after spinal cord injury in the rat. Cell Mol Neurobiol 26(7-8):1167-80.
Devine SM, Cobbs C, Jennings M, Bartholomew A, Hoffman R. 2003. Mesenchymal stem cells distribute to a wide range of tissues following systemic infusion into nonhuman primates. Blood 101(8):2999-3001.
Di Nicola M, Carlo-Stella C, Magni M, Milanesi M, Longoni PD, Matteucci P, Grisanti S, Gianni AM. 2002. Human bone marrow stromal cells suppress T-lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli. Blood 99(10):3838-43.
English K, Barry FP, Mahon BP. 2008. Murine mesenchymal stem cells suppress dendritic cell migration, maturation and antigen presentation. Immunol Lett 115(1):50-8.
Erices A, Conget P, Minguell JJ. 2000. Mesenchymal progenitor cells in human umbilical cord blood. Br J Haematol 109(1):235-42.
Grinnemo KH, Mansson A, Dellgren G, Klingberg D, Wardell E, Drvota V, Tammik C, Holgersson J, Ringden O, Sylven C and others. 2004. Xenoreactivity and engraftment of human mesenchymal stem cells transplanted into infarcted rat myocardium. J Thorac Cardiovasc Surg 127(5):1293-300.
Gronowski AM, Hilbert DM, Sheehan KC, Garotta G, Schreiber RD. 1999. Baculovirus stimulates antiviral effects in mammalian cells. J Virol 73(12):9944-51.
Hofmann C, Sandig V, Jennings G, Rudolph M, Schlag P, Strauss M. 1995. Efficient gene transfer into human hepatocytes by baculovirus vectors. Proc Natl Acad Sci U S A 92(22):10099-103.
Hsu CS, Ho YC, Wang KC, Hu YC. 2004. Investigation of optimal transduction conditions for baculovirus-mediated gene delivery into mammalian cells. Biotechnol Bioeng 88(1):42-51.
Imanishi Y, Saito A, Komoda H, Kitagawa-Sakakida S, Miyagawa S, Kondoh H, Ichikawa H, Sawa Y. 2008. Allogenic mesenchymal stem cell transplantation has a therapeutic effect in acute myocardial infarction in rats. J Mol Cell Cardiol 44(4):662-71.
Jooss K, Chirmule N. 2003. Immunity to adenovirus and adeno-associated viral vectors: implications for gene therapy. Gene Ther 10(11):955-63.
Kafri T, Morgan D, Krahl T, Sarvetnick N, Sherman L, Verma I. 1998. Cellular immune response to adenoviral vector infected cells does not require de novo viral gene expression: implications for gene therapy. Proc Natl Acad Sci U S A 95(19):11377-82.
Krampera M, Cosmi L, Angeli R, Pasini A, Liotta F, Andreini A, Santarlasci V, Mazzinghi B, Pizzolo G, Vinante F and others. 2006. Role for interferon-gamma in the immunomodulatory activity of human bone marrow mesenchymal stem cells. Stem Cells 24(2):386-98.
Le Blanc K, Ringden O. 2005. Immunobiology of human mesenchymal stem cells and future use in hematopoietic stem cell transplantation. Biol Blood Marrow Transplant 11(5):321-34.
Le Blanc K, Tammik C, Rosendahl K, Zetterberg E, Ringden O. 2003. HLA expression and immunologic properties of differentiated and undifferentiated mesenchymal stem cells. Exp Hematol 31(10):890-6.
Lundstrom K. 2003. Latest development in viral vectors for gene therapy. Trends Biotechnol 21:117-122.
Marshall E. 1999. Gene therapy death prompts review of adenovirus vector. Science 286:2244-2245.
Pieroni L, La Monica N. 2001. Towards the use of baculovirus as a gene therapy vector. Curr Opin Mol Ther 3(5):464-7.
Pieroni L, Maione D, La Monica N. 2001. In vivo gene transfer in mouse skeletal muscle mediated by baculovirus vectors. Hum Gene Ther 12(8):871-81.
Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR. 1999. Multilineage potential of adult human mesenchymal stem cells. Science 284(5411):143-7.
Plotnikov AN, Shlapakova I, Szabolcs MJ, Danilo P, Jr., Lorell BH, Potapova IA, Lu Z, Rosen AB, Mathias RT, Brink PR and others. 2007. Xenografted adult human mesenchymal stem cells provide a platform for sustained biological pacemaker function in canine heart. Circulation 116(7):706-13.
Risbud MV, Sittinger M. 2002. Tissue engineering: advances in in vitro cartilage generation. Trends Biotechnol 20(8):351-6.
Ryan JM, Barry F, Murphy JM, Mahon BP. 2007. Interferon-gamma does not break, but promotes the immunosuppressive capacity of adult human mesenchymal stem cells. Clin Exp Immunol 149(2):353-63.
Ryan JM, Barry FP, Murphy JM, Mahon BP. 2005. Mesenchymal stem cells avoid allogeneic rejection. J Inflamm (Lond) 2:8.
Saito T, Kuang JQ, Bittira B, Al-Khaldi A, Chiu RC. 2002. Xenotransplant cardiac chimera: immune tolerance of adult stem cells. Ann Thorac Surg 74(1):19-24; discussion 24.
Smith GE, Summers MD, Fraser MJ. 1983. Production of human beta interferon in insect cells infected with a baculovirus expression vector. Mol Cell Biol 3(12):2156-65.
Thomas CE, Ehrhardt A, Kay MA. 2003. Progress and problems with the use of viral vectors for gene therapy. Nature Rev. Genet. 4(5):346-358.
Tian Q, Stepaniants SB, Mao M, Weng L, Feetham MC, Doyle MJ, Yi EC, Dai H, Thorsson V, Eng J and others. 2004. Integrated genomic and proteomic analyses of gene expression in Mammalian cells. Mol Cell Proteomics 3(10):960-9.
Tse WT, Pendleton JD, Beyer WM, Egalka MC, Guinan EC. 2003. Suppression of allogeneic T-cell proliferation by human marrow stromal cells: implications in transplantation. Transplantation 75(3):389-97.
Uccelli A, Moretta L, Pistoia V. 2006. Immunoregulatory function of mesenchymal stem cells. Eur J Immunol 36(10):2566-73.
Xu XL, Tang T, Dai K, Zhu Z, Guo XE, Yu C, Lou J. 2005. Immune response and effect of adenovirus-mediated human BMP-2 gene transfer on the repair of segmental tibial bone defects in goats. Acta Orthop 76(5):637-46.