動脈粥狀硬化的病程是很多變的，有許多的因子參與其中。目前已經有很多的報導指出病原菌的感染也和動脈粥狀硬化有相關性。為了要更進一步研究疱疹病毒在動脈粥狀硬化中扮演的角色，我們建立了一個利用第四型牛疱疹病毒感染小鼠的動物模式，在這個模式中，我們將Apo-E缺陷的小鼠經鼻腔感染病毒。利用這個模式我們研究疱疹病毒在動脈粥狀硬化發展的過程中所產生的影響，結果顯示第四型牛疱疹病毒的感染會加速動脈粥狀硬化的形成，而且我們證實利用streptozotocin (STZ)這種藥物誘導Apo-E缺陷的老鼠產生糖尿病，或者是在Apo-E缺陷的老鼠施打cyclosporine A (CsA)抑制免疫功能，也都會促進動脈粥狀硬化更嚴重。因此，在這個實驗中，我們想要更進一步研究免疫力低下對於第四型牛疱疹病毒感染引發動脈粥狀硬化過程中的影響，我們的結果顯示感染病毒後又加以誘導糖尿病或施打CsA並不會影響血漿中膽固醇的含量。然而在這兩種狀況下，都可以使動脈粥狀硬化的形成加速，以及降低淋巴細胞增生的能力。可見免疫系統也會影響動脈粥狀硬化的形成，所以我們利用了流式細胞儀和免疫組織染色的方法偵測免疫細胞的分布，細胞激素的分泌情形，以及小鼠之第一型輔助性T細胞和第二型輔助性T細胞之間的比值。結果感染病毒後又加以誘導糖尿病或施打CsA會對第一型輔助性T細胞/第二型輔助性T細胞之間的平衡，產生輕微調整的趨勢。總結，免疫力低下在Apo-E缺陷的小鼠身上會去促進第四型牛疱疹病毒感染後引發的動脈粥狀硬化形成的程度，而第一型輔助性T細胞/第二型輔助性T細胞之間的轉換可能會影響動脈粥狀硬化形成中的發炎和免疫反應。

The pathogenesis of atherosclerosis is diverse and involves many factors. Involvement of infectious agents in the pathogenesis of atherosclerosis has been suggested in different reports. To facilitate the investigation of the role of herpesvirus in atherosclerosis, we established an animal model in murine by subclinical infection with bovine herpes virus type 4 (BHV-4). In the model, the apoE (-/-) mice were infected with virus by intranasal inoculation. With this model we studied the effects of herpesviral infection on the development of atherosclerosis. The result demonstrated that infection of BHV-4 could accelerate atherogenic process. Furthermore, we demonstrated that severer atherosclerosis occured in Streptozotocin (STZ)-induced DM or cyclosporine A (CsA)-treated apoE(-/-) mice. Therefore, in this study, we aim to investigate the effect of immunocompromise in BHV-4 infection induced atherosclerosis. Our results showed there is no significant difference in the cholesterol levels of mice treated with BHV-4 in combination with STZ-induced DM or CsA-treatment. However, accelerated atherosclerotic lesions and reduced lymphocyte proliferation were observed in both treated groups. Since immune system may affect the development of atherosclerosis, we utilized flow cytometry and immunohistochemistry to analyze the immune cell subtyping, cytokine release, and the ratios of Th1/Th2 in the mice. We found that BHV-4 infection in combination with STZ-induced DM or CsA-treated may slightly modulate Th1/Th2 balance. In conclusion, immunocompromise might promote the formation of atherosclerotic lesions in apoE(-/-) mice with BHV-4 infection. The switch of Th1 and Th2 balance may influence the inflammatory and immune processes occur in atherosclerosis plaque.

Abbas, A. K., Murphy, K. M., and Sher, A.: Functional diversity of helper T lymphocytes. Nature 383: 787-93, 1996
Adam, E., Melnick, J. L., Probtsfield, J. L., Petrie, B. L., Burek, J., Bailey, K. R., McCollum, C. H., and DeBakey, M. E.: High levels of cytomegalovirus antibody in patients requiring vascular surgery for atherosclerosis. Lancet 2: 291-3, 1987
Alber, D. G., Powell, K. L., Vallance, P., Goodwin, D. A., and Grahame-Clarke, C.: Herpesvirus infection accelerates atherosclerosis in the apolipoprotein E-deficient mouse. Circulation 102: 779-85, 2000
Alber, D. G., Vallance, P., and Powell, K. L.: Enhanced atherogenesis is not an obligatory response to systemic herpesvirus infection in the apoE-deficient mouse: comparison of murine gamma-herpesvirus-68 and herpes simplex virus-1. Arterioscler Thromb Vasc Biol 22: 793-8, 2002
Amos, A. F., McCarty, D. J., and Zimmet, P.: The rising global burden of diabetes and its complications: estimates and projections to the year 2010. Diabet Med 14 Suppl 5: S1-85, 1997
Assenmacher, M., Schmitz, J., and Radbruch, A.: Flow cytometric determination of cytokines in activated murine T helper lymphocytes: expression of interleukin-10 in interferon-gamma and in interleukin-4-expressing cells. Eur J Immunol 24: 1097-101, 1994
Ballantyne, C. M., Podet, E. J., Patsch, W. P., Harati, Y., Appel, V., Gotto, A. M., Jr., and Young, J. B.: Effects of cyclosporine therapy on plasma lipoprotein levels. Jama 262: 53-6, 1989
Bruggeman, C. A.: Cytomegalovirus and latency: an overview. Virchows Arch B Cell Pathol Incl Mol Pathol 64: 325-33, 1993
Bublot, M., Lomonte, P., Lequarre, A. S., Albrecht, J. C., Nicholas, J., Fleckenstein, B., Pastoret, P. P., and Thiry, E.: Genetic relationships between bovine herpesvirus 4 and the gammaherpesviruses Epstein-Barr virus and herpesvirus saimiri. Virology 190: 654-65, 1992
Carr, M. E.: Diabetes mellitus: a hypercoagulable state. J Diabetes Complications 15: 44-54, 2001
Ceriello, A., Giacomello, R., Stel, G., Motz, E., Taboga, C., Tonutti, L., Pirisi, M., Falleti, E., and Bartoli, E.: Hyperglycemia-induced thrombin formation in diabetes. The possible role of oxidative stress. Diabetes 44: 924-8, 1995
Ceriello, A., Giugliano, D., Quatraro, A., Dello Russo, P., and Torella, R.: Blood glucose may condition factor VII levels in diabetic and normal subjects. Diabetologia 31: 889-91, 1988
Ceriello, A., Giugliano, D., Quatraro, A., Marchi, E., Barbanti, M., and Lefebvre, P.: Evidence for a hyperglycaemia-dependent decrease of antithrombin III-thrombin complex formation in humans. Diabetologia 33: 163-7, 1990
Citterio, F. and Kahan, B. D.: Effects of cyclosporin on nuclear function of transplant recipient lymphocytes. Transplant Proc 21: 1465-6, 1989
Creager, M. A., Luscher, T. F., Cosentino, F., and Beckman, J. A.: Diabetes and vascular disease: pathophysiology, clinical consequences, and medical therapy: Part I. Circulation 108: 1527-32, 2003
Dansky, H. M., Charlton, S. A., Harper, M. M., and Smith, J. D.: T and B lymphocytes play a minor role in atherosclerotic plaque formation in the apolipoprotein E-deficient mouse. Proc Natl Acad Sci U S A 94: 4642-6, 1997
de Boer, O. J., van der Wal, A. C., and Becker, A. E.: Atherosclerosis, inflammation, and infection. J Pathol 190: 237-43, 2000
Diepersloot, R. J., Bouter, K. P., Beyer, W. E., Hoekstra, J. B., and Masurel, N.: Humoral immune response and delayed type hypersensitivity to influenza vaccine in patients with diabetes mellitus. Diabetologia 30: 397-401, 1987
Diepersloot, R. J., Bouter, K. P., van Beek, R., Lucas, C. J., Masurel, N., and Erkelens, D. W.: Cytotoxic T-cell response to influenza A subunit vaccine in patients with type 1 diabetes mellitus. Neth J Med 35: 68-75, 1989
Egyed, L., Ballagi-Pordany, A., Bartha, A., and Belak, S.: Studies of in vivo distribution of bovine herpesvirus type 4 in the natural host. J Clin Microbiol 34: 1091-5, 1996
Emeson, E. E. and Shen, M. L.: Accelerated atherosclerosis in hyperlipidemic C57BL/6 mice treated with cyclosporin A. Am J Pathol 142: 1906-15, 1993
Fabricant, C. G.: Atherosclerosis: the consequence of infection with a herpesvirus. Adv Vet Sci Comp Med 30: 39-66, 1985
Fabricant, C. G., Fabricant, J., Minick, C. R., and Litrenta, M. M.: Herpesvirus-induced atherosclerosis in chickens. Fed Proc 42: 2476-9, 1983
Fernandes, G., Handwerger, B. S., Yunis, E. J., and Brown, D. M.: Immune response in the mutant diabetic C57BL/Ks-dt+ mouse. Discrepancies between in vitro and in vivo immunological assays. J Clin Invest 61: 243-50, 1978
Ferns, G., Reidy, M., and Ross, R.: Vascular effects of cyclosporine A in vivo and in vitro. Am J Pathol 137: 403-13, 1990
Friedman, H. M., Macarak, E. J., MacGregor, R. R., Wolfe, J., and Kefalides, N. A.: Virus infection of endothelial cells. J Infect Dis 143: 266-73, 1981
Frostegard, J., Ulfgren, A. K., Nyberg, P., Hedin, U., Swedenborg, J., Andersson, U., and Hansson, G. K.: Cytokine expression in advanced human atherosclerotic plaques: dominance of pro-inflammatory (Th1) and macrophage-stimulating cytokines. Atherosclerosis 145: 33-43, 1999
Granelli-Piperno, A.: Lymphokine gene expression in vivo is inhibited by cyclosporin A. J Exp Med 171: 533-44, 1990
Grattan, M. T., Moreno-Cabral, C. E., Starnes, V. A., Oyer, P. E., Stinson, E. B., and Shumway, N. E.: Cytomegalovirus infection is associated with cardiac allograft rejection and atherosclerosis. Jama 261: 3561-6, 1989
Gupta, S., Pablo, A. M., Jiang, X., Wang, N., Tall, A. R., and Schindler, C.: IFN-gamma potentiates atherosclerosis in ApoE knock-out mice. J Clin Invest 99: 2752-61, 1997
Haffner, S. M., Lehto, S., Ronnemaa, T., Pyorala, K., and Laakso, M.: Mortality from coronary heart disease in subjects with type 2 diabetes and in nondiabetic subjects with and without prior myocardial infarction. N Engl J Med 339: 229-34, 1998
Hansson, G. K.: Cell-mediated immunity in atherosclerosis. Curr Opin Lipidol 8: 301-11, 1997
Hendrix, M. G., Dormans, P. H., Kitslaar, P., Bosman, F., and Bruggeman, C. A.: The presence of cytomegalovirus nucleic acids in arterial walls of atherosclerotic and nonatherosclerotic patients. Am J Pathol 134: 1151-7, 1989
Hsich, E., Zhou, Y. F., Paigen, B., Johnson, T. M., Burnett, M. S., and Epstein, S. E.: Cytomegalovirus infection increases development of atherosclerosis in Apolipoprotein-E knockout mice. Atherosclerosis 156: 23-8, 2001
Huber, S. A., Sakkinen, P., David, C., Newell, M. K., and Tracy, R. P.: T helper-cell phenotype regulates atherosclerosis in mice under conditions of mild hypercholesterolemia. Circulation 103: 2610-6, 2001
Jonasson, L., Holm, J., and Hansson, G. K.: Cyclosporin A inhibits smooth muscle proliferation in the vascular response to injury. Proc Natl Acad Sci U S A 85: 2303-6, 1988
Kunjathoor, V. V., Wilson, D. L., and LeBoeuf, R. C.: Increased atherosclerosis in streptozotocin-induced diabetic mice. J Clin Invest 97: 1767-73, 1996
Laurat, E., Poirier, B., Tupin, E., Caligiuri, G., Hansson, G. K., Bariety, J., and Nicoletti, A.: In vivo downregulation of T helper cell 1 immune responses reduces atherogenesis in apolipoprotein E-knockout mice. Circulation 104: 197-202, 2001
Libby, P.: Changing concepts of atherogenesis. J Intern Med 247: 349-58, 2000
Lin, T. M., Chen, W. J., Chen, H. Y., Wang, P. W., and Eng, H. L.: Increased incidence of cytomegalovirus but not Chlamydia pneumoniae in atherosclerotic lesions of arteries of lower extremities from patients with diabetes mellitus undergoing amputation. J Clin Pathol 56: 429-32, 2003
Lin, T. M., Jiang, M. J., Eng, H. L., Shi, G. Y., Lai, L. C., Huang, B. J., Huang, K. Y., and Wu, H. L.: Experimental infection with bovine herpesvirus-4 enhances atherosclerotic process in rabbits. Lab Invest 80: 3-11, 2000
Lin, T. M., Shi, G. Y., Tsai, C. F., Su, H. J., Guo, Y. L., and Wu, H. L.: Susceptibility of endothelial cells to bovine herpesvirus type 4 (BHV-4). J Virol Methods 63: 219-25, 1997
Lustig, S., Stern, N., Lee, D. B., Berger, M. E., Eggena, P., Balabanian, M., and Golub, M. S.: Mechanisms of cyclosporine A hypertension. Transplant Proc 19: 1262-4, 1987
Maggi, E., Parronchi, P., Manetti, R., Simonelli, C., Piccinni, M. P., Rugiu, F. S., De Carli, M., Ricci, M., and Romagnani, S.: Reciprocal regulatory effects of IFN-gamma and IL-4 on the in vitro development of human Th1 and Th2 clones. J Immunol 148: 2142-7, 1992
Muller, A., Schott-Ohly, P., Dohle, C., and Gleichmann, H.: Differential regulation of Th1-type and Th2-type cytokine profiles in pancreatic islets of C57BL/6 and BALB/c mice by multiple low doses of streptozotocin. Immunobiology 205: 35-50, 2002
Nguyen, D. T., Eskandai, M. K., DeForge, L. E., Raiford, C. L., Strieter, R. M., Kunkel, S. L., and Remick, D. G.: Cyclosporin a modulation of tumor necrosis factor gene expression and effects in vitro and in vivo. J Immunol 144: 3822-8, 1990
Nieto, F. J.: Viruses and atherosclerosis: a critical review of the epidemiologic evidence. Am Heart J 138: S453-60, 1999
Nieto, F. J., Adam, E., Sorlie, P., Farzadegan, H., Melnick, J. L., Comstock, G. W., and Szklo, M.: Cohort study of cytomegalovirus infection as a risk factor for carotid intimal-medial thickening, a measure of subclinical atherosclerosis. Circulation 94: 922-7, 1996
O'Garra, A.: Cytokines induce the development of functionally heterogeneous T helper cell subsets. Immunity 8: 275-83, 1998
Osorio, F. A., Galeota-Wheeler, J. A., Reed, D. E., and Lopez, O.: About the cellular tropism of the gammaherpesvirus bovine herpesvirus type 4. J Clin Microbiol 34: 3253-4, 1996
Osorio, F. A. and Reed, D. E.: Experimental inoculation of cattle with bovine herpesvirus-4: evidence for a lymphoid-associated persistent infection. Am J Vet Res 44: 975-80, 1983
Paludan, S. R.: Interleukin-4 and interferon-gamma: the quintessence of a mutual antagonistic relationship. Scand J Immunol 48: 459-68, 1998
Petrie, B. L., Melnick, J. L., Adam, E., Burek, J., McCollum, C. H., and DeBakey, M. E.: Nucleic acid sequences of cytomegalovirus in cells cultured from human arterial tissue. J Infect Dis 155: 158-9, 1987
Ross, R.: The pathogenesis of atherosclerosis--an update. N Engl J Med 314: 488-500, 1986
Ross, R.: The pathogenesis of atherosclerosis: a perspective for the 1990s. Nature 362: 801-9, 1993
Ross, R.: Atherosclerosis--an inflammatory disease. N Engl J Med 340: 115-26, 1999
Ross, R. and Glomset, J. A.: The pathogenesis of atherosclerosis (first of two parts). N Engl J Med 295: 369-77, 1976
Salameh, A., Zinn, M., and Dhein, S.: High D-glucose induces alterations of endothelial cell structure in a cell-culture model. J Cardiovasc Pharmacol 30: 182-90, 1997
Song, L., Leung, C., and Schindler, C.: Lymphocytes are important in early atherosclerosis. J Clin Invest 108: 251-9, 2001
Tesfamariam, B., Brown, M. L., and Cohen, R. A.: Elevated glucose impairs endothelium-dependent relaxation by activating protein kinase C. J Clin Invest 87: 1643-8, 1991
Vercellotti, G. M.: Overview of infections and cardiovascular diseases. J Allergy Clin Immunol 108: S117-20, 2001
Vliegen, I., Stassen, F., Grauls, G., Blok, R., and Bruggeman, C.: MCMV infection increases early T-lymphocyte influx in atherosclerotic lesions in apoE knockout mice. J Clin Virol 25 Suppl 2: S159-71, 2002
Yamashiroya, H. M., Ghosh, L., Yang, R., and Robertson, A. L., Jr.: Herpesviridae in the coronary arteries and aorta of young trauma victims. Am J Pathol 130: 71-9, 1988
Zhou, X., Paulsson, G., Stemme, S., and Hansson, G. K.: Hypercholesterolemia is associated with a T helper (Th) 1/Th2 switch of the autoimmune response in atherosclerotic apo E-knockout mice. J Clin Invest 101: 1717-25, 1998
Zoja, C., Furci, L., Ghilardi, F., Zilio, P., Benigni, A., and Remuzzi, G.: Cyclosporin-induced endothelial cell injury. Lab Invest 55: 455-62, 1986
王昱竺 利用動物模式探討宿主免疫力低下與病毒感染加速動脈硬化的關係 國立成功大學醫事技術學系學士論文集 3:1-26 2002