臺灣博碩士論文加值系統

腸病毒71型感染通常與手足口病有關，並可能導致嚴重的神經系統疾病甚至死亡。雖然之前的研究建立了許多動物模式來研究腸病毒71型的致病機轉，但是目前卻缺乏一種能藉口服感染腸病毒71型臨床株的動物模式。因此，我們開發了一種能表達人類清道夫受體B類成員2的轉基因小鼠。我們將帶有FLAG標記的人類清道夫受體B類成員2第三外顯子到第12外顯子互補脫氧核醣核酸片段插入含有小鼠清道夫受體B類成員2基因的細菌人工染色體克隆中，並將得到的轉基因用於建立表達嵌合受體的轉基因小鼠。藉由口餵感染腸病毒71型臨床株的轉基因小鼠，顯示出如共濟失調和麻痺的神經症狀，嚴重者則會死亡。受感染的轉基因小鼠的病理學特徵，則類似於人類患者的腦脊髓炎，並且伴有小膠質細胞的活化。剔除小膠質細胞的轉基因小鼠腦切片會增強腸病毒複製的效果，而預先活化小膠質細胞的轉基因小鼠腦切片則會抑制病毒的複製。實驗結果表明，小膠質細胞具有抗腸病毒71型感染的作用。該轉基因小鼠鼠提供了類似於天然感染的模型，適用於研究腸病毒71型的發病機理和評估疫苗或其他抗病毒藥物的功效。

Enterovirus 71 (EV71) has caused several outbreaks of hand foot and mouth disease (HFMD) in regions such as Japan, Taiwan, and China. Infection with EV71 may also cause severe neurologic diseases, such as acute encephalitis and acute flaccid paralysis, and it may even be fatal. To study the neuropathogenicity of EV71, experiments using animal model are essential. We therefore developed a transgenic (Tg) mouse model expressing the EV71 receptor, human scavenger receptor class B2 (hSCARB2), which is highly similar to murine endogenous mSCARB2 protein. We generated a chimeric receptor-expressing Tg mouse model, susceptible to oral infection with EV71, by inserting a FLAG-tagged SCARB2 complementary DNA fragment composed of exons 3–12 in a murine SCARB2 gene-containing bacterial artificial chromosome clone. The Tg mice, oral-infected with clinical isolates of EV71, showed neurologic symptoms, such as ataxia, paralysis, and fatality, and exhibited age-dependent susceptibility similar to that in humans. Viral infection was accompanied by the activation of microglia. Clodronate treatment of brain slices from the Tg mice enhanced viral replication, while lipopolysaccharide treatment significantly inhibited it, thus suggesting an antiviral role for microglia during EV71 infection. In summary, the Tg mouse model mimics natural infections and can be used to study the pathogenesis of EV71 and to assess the efficacy of vaccines or other antiviral drugs.

Table of contents
Recommendation Letter from the Thesis Advisor
Dissertation Oral Defense Committee Certification
Chinese Abstract.....iii
English Abstract.....iv
Table of Contents.....v
List of figures .....vii
List of table.....ix
Chapter 1. Introduction .....1
1.1 General features of EV71.....1
1.2 Clinical features of EV71 infection in humans.....2
1.3 Host Receptors for EV71 .....3
1.4 In vivo animal models used for EV71 infection.....7
1.5 Role of microglia in CNS infections.....10
Chapter 2. Materials and methods.....13
Chapter 3. Results.....22
3.1 Generation of a Tg murine model that expresses mSCARB2/hSCARB2 chimeric receptors.....22
3.2 Expression profile of mSCARB2/hSCARB2 chimeric protein in a Tg murine model.....22
3.3 Increased susceptibility of Tg mice to EV71 infection via the intragastric route.....24
3.4 EV71 replication in various tissues in Tg mice..... 26
3.5 Increased cytokines and chemokines in the brains of EV71-infected Tg mice.....28
3.6 Microglial activation in the brains of EV71-infected Tg mice.....29
Chapter 4. Discussion.....33
Reference.....39
Figures.....48
Table.....80

List of figures
Figure 1. Generation of Scarb2-SCARB2 BAC Tg mice......48
Figure 2. Tissue distribution of mSCARB2/hSCARB2 chimeric protein in the Tg and non-Tg mice.....49
Figure 3. Expression profiles of hSCARB2 in the central nervous system of Tg mouse......50
Figure 4. Expression profiles of hSCARB2 in the digestive system of Tg mouse.....52
Figure 5. Expression profiles of hSCARB2 in the Tg mouse.....54
Figure 6. Mortality and neurological symptoms observed in EV71 intragastrically infected Tg mice with different viral doses.....56
Figure 7. Mortality and neurological symptoms observed in EV71 intragastrically infected Tg mice with different mouse ages.....57
Figure 8. Mortality and neurological symptoms observed in EV71 intragastrically infected Tg mice with different clinical isolate strains.....58
Figure 9. The Tg mice are susceptible to EV71 infection under different inoculation routes.....59
Figure 10. The Tg mice are susceptible to oral CVA16 infection.....60
Figure 11. Replication of EV71 in the nerves system of orally infected Tg mice.....61
Figure 12. Replication of EV71 in the hind legs of orally infected Tg mice......64
Figure 13. Viral titers in the CNS of the infected mice.....66
Figure 14. Viral titers in the gastrointestinal tract of the infected mice.....67
Figure 15. Viral titers in skeletal muscle of the infected mice 68
Figure 16. Expression of pro-inflammatory cytokines in the brains of EV71-infected mice.....69
Figure 17. Expression of chemokine and EV71 genomic RNA in the brains of EV71-infected mice.....70
Figure 18. Microglial activation in the brain of the infected mice.....71
Figure 19. Microglial activation in the spinal cord of the infected mice.....72
Figure 20. Increases in microglia number in infected Tg mice with symptoms......73
Figure 21. Antiviral response of the activated microglia.....74
Figure 22. Expression of pro-inflammatory cytokines in microglia from the EV71-infected Tg mice.....76
Figure 23. Expression of chemokine and EV71 genomic RNA in the microglia of EV71-infected Tg mice.....78

List of table
Table 1. A list of primers used in this study......80

1. Chung PW, Huang YC, Chang LY, Lin TY, Ning HC. 2001. Duration of enterovirus shedding in stool. Journal of Microbiology, Immunology and Infection 34:167-170.
2. He Y, Ong KC, Gao Z, Zhao X, Anderson VM, McNutt MA, Wong KT, Lu M. 2014. Tonsillar crypt epithelium is an important extra-central nervous system site for viral replication in EV71 encephalomyelitis. The American Journal of Pathology 184:714-720.
3. Cheng HY, Huang YC, Yen TY, Hsia SH, Hsieh YC, Li CC, Chang LY, Huang LM. 2014. The correlation between the presence of viremia and clinical severity in patients with enterovirus 71 infection: a multi-center cohort study. BMC Infectious Diseases 14:417.
4. Solomon T, Lewthwaite P, Perera D, Cardosa MJ, McMinn P, Ooi MH. 2010. Virology, epidemiology, pathogenesis, and control of enterovirus 71. The Lancet Infectious Diseases 10:778-790.
5. Weng KF, Chen LL, Huang PN, Shih SR. 2010. Neural pathogenesis of enterovirus 71 infection. Microbes and Infection 12:505-510.
6. Yang F, Ren L, Xiong Z, Li J, Xiao Y, Zhao R, He Y, Bu G, Zhou S, Wang J, Qi J. 2009. Enterovirus 71 outbreak in the People's Republic of China in 2008. Journal of Clinical Microbiology 47:2351-2352.
7. Yang F, Zhang T, Hu Y, Wang X, Du J, Li Y, Sun S, Sun X, Li Z, Jin Q. 2011. Survey of enterovirus infections from hand, foot and mouth disease outbreak in China, 2009. Virology journal 8:508.
8. Mao LX, Wu B, Bao WX, Han FA, Xu L, Ge QJ, Yang J, Yuan ZH, Miao CH, Huang XX, Zhang C, Xu H. 2010. Epidemiology of hand, foot, and mouth disease and genotype characterization of Enterovirus 71 in Jiangsu, China. Journal of Clinical Virology 49:100-104.
9. Shieh WJ, Jung SM, Hsueh C, Kuo TT, Mounts A, Parashar U, Yang CF, Guarner J, Ksiazek TG, Dawson J, Goldsmith C, Chang GJ, Oberste SM, Pallansch MA, Anderson LJ, Zaki SR. 2001. Pathologic studies of fatal cases in outbreak of hand, foot, and mouth disease, Taiwan. Emerging Infectious Diseases 7:146-148.
10. Yang Y, Wang H, Gong E, Du J, Zhao X, McNutt MA, Wang S, Zhong Y, Gao Z, Zheng J. 2009. Neuropathology in 2 cases of fatal enterovirus type 71 infection from a recent epidemic in the People's Republic of China: a histopathologic, immunohistochemical, and reverse transcription polymerase chain reaction study. Human Pathology 40:1288-1295.
11. Lin TY, Hsia SH, Huang YC, Wu CT, Chang LY. 2003. Proinflammatory cytokine reactions in enterovirus 71 infections of the central nervous system. Clinical Infectious Diseases 36:269-274.
12. Lum LC, Wong KT, Lam SK, Chua KB, Goh AY, Lim WL, Ong BB, Paul G, AbuBakar S, Lambert M. 1998. Fatal enterovirus 71 encephalomyelitis. The Journal of Pediatrics 133:795-798.
13. Lin TY, Chang LY, Huang YC, Hsu KH, Chiu CH, Yang KD. 2002. Different proinflammatory reactions in fatal and non-fatal enterovirus 71 infections: implications for early recognition and therapy. Acta Paediatrica 91:632-635.
14. Ye N, Gong X, Pang LL, Gao WJ, Zhang YT, Li XL, Liu N, Li DD, Jin Y, Duan ZJ. 2015. Cytokine responses and correlations thereof with clinical profiles in children with enterovirus 71 infections. BMC Infectious Diseases 15:225.
15. Griffiths MJ, Ooi MH, Wong SC, Mohan A, Podin Y, Perera D, Chieng CH, Tio PH, Cardosa MJ, Solomon T. 2012. In enterovirus 71 encephalitis with cardio-respiratory compromise, elevated interleukin 1beta, interleukin 1 receptor antagonist, and granulocyte colony-stimulating factor levels are markers of poor prognosis. The Journal of Infectious Diseases 206:881-892.
16. Yamayoshi S, Yamashita Y, Li J, Hanagata N, Minowa T, Takemura T, Koike S. 2009. Scavenger receptor B2 is a cellular receptor for enterovirus 71. Nature Medicine 15:798-801.
17. Nishimura Y, Shimojima M, Tano Y, Miyamura T, Wakita T, Shimizu H. 2009. Human P-selectin glycoprotein ligand-1 is a functional receptor for enterovirus 71. Nature Medicine 15:794-797.
18. Yang SL, Chou YT, Wu CN, Ho MS. 2011. Annexin II binds to capsid protein VP1 of enterovirus 71 and enhances viral infectivity. Journal of virology 85:11809-11820.
19. Yang B, Chuang H, Yang KD. 2009. Sialylated glycans as receptor and inhibitor of enterovirus 71 infection to DLD-1 intestinal cells. Virology journal 6:141.
20. Tan CW, Poh CL, Sam IC, Chan YF. 2013. Enterovirus 71 uses cell surface heparan sulfate glycosaminoglycan as an attachment receptor. Journal of virology 87:611-620.
21. Du N, Cong H, Tian H, Zhang H, Zhang W, Song L, Tien P. 2014. Cell surface vimentin is an attachment receptor for enterovirus 71. Journal of virology 88:5816-5833.
22. Su PY, Wang YF, Huang SW, Lo YC, Wang YH, Wu SR, Shieh DB, Chen SH, Wang JR, Lai MD, Chang CF. 2015. Cell surface nucleolin facilitates enterovirus 71 binding and infection. Journal of virology 89:4527-4538.
23. Srivastava M, Pollard HB. 1999. Molecular dissection of nucleolin's role in growth and cell proliferation: new insights. The FASEB Journal 13:1911-1922.
24. Alvarez Losada S, Canto-Nogues C, Munoz-Fernandez MA. 2002. A new possible mechanism of human immunodeficiency virus type 1 infection of neural cells. Neurobiology of Disease 11:469-478.
25. Tayyari F, Marchant D, Moraes TJ, Duan W, Mastrangelo P, Hegele RG. 2011. Identification of nucleolin as a cellular receptor for human respiratory syncytial virus. Nature Medicine 17:1132-1135.
26. Krust B, Vienet R, Cardona A, Rougeot C, Jacotot E, Callebaut C, Guichard G, Briand JP, Grognet JM, Hovanessian AG, Edelman L. 2001. The anti-HIV pentameric pseudopeptide HB-19 is preferentially taken up in vivo by lymphoid organs where it forms a complex with nucleolin. Proceedings of the National Academy of Sciences of the United States of America 98:14090-14095.
27. Alete DE, Weeks ME, Hovanession AG, Hawadle M, Stoker AW. 2006. Cell surface nucleolin on developing muscle is a potential ligand for the axonal receptor protein tyrosine phosphatase-sigma. The FEBS Journal 273:4668-4681.
28. Varki NM, Varki A. 2007. Diversity in cell surface sialic acid presentations: implications for biology and disease. Laboratory investigation 87:851-857.
29. Nilsson EC, Jamshidi F, Johansson SM, Oberste MS, Arnberg N. 2008. Sialic acid is a cellular receptor for coxsackievirus A24 variant, an emerging virus with pandemic potential. Journal of virology 82:3061-3068.
30. Suzuki Y. 2009. The highly pathogenic avian influenza H5N1 - initial molecular signals for the next influenza pandemic. Chang Gung Medical Journal 32:258-263.
31. Su PY, Liu YT, Chang HY, Huang SW, Wang YF, Yu CK, Wang JR, Chang CF. 2012. Cell surface sialylation affects binding of enterovirus 71 to rhabdomyosarcoma and neuroblastoma cells. BMC Microbiology 12:162.
32. Merilahti P, Karelehto E, Susi P. 2016. Role of Heparan Sulfate in Cellular Infection of Integrin-Binding Coxsackievirus A9 and Human Parechovirus 1 Isolates. PLoS One 11:e0147168.
33. Zhang X, Shi J, Ye X, Ku Z, Zhang C, Liu Q, Huang Z. 2017. Coxsackievirus A16 utilizes cell surface heparan sulfate glycosaminoglycans as its attachment receptor. Emerging Microbes & Infections 6:e65.
34. Tseligka ED, Sobo K, Stoppini L, Cagno V, Abdul F, Piuz I, Meylan P, Huang S, Constant S, Tapparel C. 2018. A VP1 mutation acquired during an enterovirus 71 disseminated infection confers heparan sulfate binding ability and modulates ex vivo tropism. PLoS Pathog 14:e1007190.
35. Donato R, Russo-Marie F. 1999. The annexins: structure and functions. Cell Calcium 26:85-89.
36. Kim J, Hajjar KA. 2002. Annexin II: a plasminogen-plasminogen activator co-receptor. Frontiers in Bioscience 7:d341-348.
37. Das S, Ravi V, Desai A. 2011. Japanese encephalitis virus interacts with vimentin to facilitate its entry into porcine kidney cell line. Virus research 160:404-408.
38. Nishimura Y, Wakita T, Shimizu H. 2010. Tyrosine sulfation of the amino terminus of PSGL-1 is critical for enterovirus 71 infection. PLoS Pathog 6:e1001174.
39. Nishimura Y, Lee H, Hafenstein S, Kataoka C, Wakita T, Bergelson JM, Shimizu H. 2013. Enterovirus 71 binding to PSGL-1 on leukocytes: VP1-145 acts as a molecular switch to control receptor interaction. PLoS Pathog 9:e1003511.
40. Laszik Z, Jansen PJ, Cummings RD, Tedder TF, McEver RP, Moore KL. 1996. P-selectin glycoprotein ligand-1 is broadly expressed in cells of myeloid, lymphoid, and dendritic lineage and in some nonhematopoietic cells. Blood 88:3010-3021.
41. Eskelinen EL, Tanaka Y, Saftig P. 2003. At the acidic edge: emerging functions for lysosomal membrane proteins. Trends in cell biology 13:137-145.
42. Yamayoshi S, Iizuka S, Yamashita T, Minagawa H, Mizuta K, Okamoto M, Nishimura H, Sanjoh K, Katsushima N, Itagaki T, Nagai Y, Fujii K, Koike S. 2012. Human SCARB2-dependent infection by coxsackievirus A7, A14, and A16 and enterovirus 71. Journal of virology 86:5686-5696.
43. Yamayoshi S, Koike S. 2011. Identification of a human SCARB2 region that is important for enterovirus 71 binding and infection. Journal of virology 85:4937-4946.
44. Hashimoto I, Hagiwara A. 1982. Pathogenicity of a poliomyelitis-like disease in monkeys infected orally with enterovirus 71: a model for human infection. Neuropathology and applied neurobiology 8:149-156.
45. Chumakov M, Voroshilova M, Shindarov L, Lavrova I, Gracheva L, Koroleva G, Vasilenko S, Brodvarova I, Nikolova M, Gyurova S, Gacheva M, Mitov G, Ninov N, Tsylka E, Robinson I, Frolova M, Bashkirtsev V, Martiyanova L, Rodin V. 1979. Enterovirus 71 isolated from cases of epidemic poliomyelitis-like disease in Bulgaria. Archives of virology 60:329-340.
46. Hashimoto I, Hagiwara A, Kodama H. 1978. Neurovirulence in cynomolgus monkeys of enterovirus 71 isolated from a patient with hand, foot and mouth disease. Archives of virology 56:257-261.
47. Arita M, Shimizu H, Nagata N, Ami Y, Suzaki Y, Sata T, Iwasaki T, Miyamura T. 2005. Temperature-sensitive mutants of enterovirus 71 show attenuation in cynomolgus monkeys. Journal of General Virology 86:1391-1401.
48. Arita M, Nagata N, Iwata N, Ami Y, Suzaki Y, Mizuta K, Iwasaki T, Sata T, Wakita T, Shimizu H. 2007. An attenuated strain of enterovirus 71 belonging to genotype a showed a broad spectrum of antigenicity with attenuated neurovirulence in cynomolgus monkeys. Journal of virology 81:9386-9395.
49. Yu CK, Chen CC, Chen CL, Wang JR, Liu CC, Yan JJ, Su IJ. 2000. Neutralizing antibody provided protection against enterovirus type 71 lethal challenge in neonatal mice. Journal of Biomedical Science 7:523-528.
50. Wang YF, Chou CT, Lei HY, Liu CC, Wang SM, Yan JJ, Su IJ, Wang JR, Yeh TM, Chen SH, Yu CK. 2004. A mouse-adapted enterovirus 71 strain causes neurological disease in mice after oral infection. Journal of virology 78:7916-7924.
51. Ong KC, Badmanathan M, Devi S, Leong KL, Cardosa MJ, Wong KT. 2008. Pathologic characterization of a murine model of human enterovirus 71 encephalomyelitis. Journal of Neuropathology & Experimental Neurology 67:532-542.
52. Chua BH, Phuektes P, Sanders SA, Nicholls PK, McMinn PC. 2008. The molecular basis of mouse adaptation by human enterovirus 71. The Journal of general virology 89:1622-1632.
53. Liao CC, Liou AT, Chang YS, Wu SY, Chang CS, Lee CK, Kung JT, Tu PH, Yu YY, Lin CY, Lin JS, Shih C. 2014. Immunodeficient mouse models with different disease profiles by in vivo infection with the same clinical isolate of enterovirus 71. Journal of virology 88:12485-12499.
54. Arita M, Ami Y, Wakita T, Shimizu H. 2008. Cooperative effect of the attenuation determinants derived from poliovirus sabin 1 strain is essential for attenuation of enterovirus 71 in the NOD/SCID mouse infection model. Journal of virology 82:1787-1797.
55. Khong WX, Yan B, Yeo H, Tan EL, Lee JJ, Ng JK, Chow VT, Alonso S. 2012. A non-mouse-adapted enterovirus 71 (EV71) strain exhibits neurotropism, causing neurological manifestations in a novel mouse model of EV71 infection. Journal of virology 86:2121-2131.
56. Lin YW, Yu SL, Shao HY, Lin HY, Liu CC, Hsiao KN, Chitra E, Tsou YL, Chang HW, Sia C, Chong P, Chow YH. 2013. Human SCARB2 transgenic mice as an infectious animal model for enterovirus 71. PLoS One 8:e57591.
57. Fujii K, Nagata N, Sato Y, Ong KC, Wong KT, Yamayoshi S, Shimanuki M, Shitara H, Taya C, Koike S. 2013. Transgenic mouse model for the study of enterovirus 71 neuropathogenesis. Proceedings of the National Academy of Sciences of the United States of America 110:14753-14758.
58. Yu P, Gao Z, Zong Y, Bao L, Xu L, Deng W, Li F, Lv Q, Gao Z, Xu Y, Yao Y, Qin C. 2014. Histopathological features and distribution of EV71 antigens and SCARB2 in human fatal cases and a mouse model of enterovirus 71 infection. Virus research 189:121-132.
59. Gomez Perdiguero E, Klapproth K, Schulz C, Busch K, Azzoni E, Crozet L, Garner H, Trouillet C, de Bruijn MF, Geissmann F, Rodewald HR. 2015. Tissue-resident macrophages originate from yolk-sac-derived erythro-myeloid progenitors. Nature 518:547-551.
60. Kalkman HO, Feuerbach D. 2016. Antidepressant therapies inhibit inflammation and microglial M1-polarization. Pharmacology & Therapeutics 163:82-93.
61. Varin A, Gordon S. 2009. Alternative activation of macrophages: immune function and cellular biology. Immunobiology 214:630-641.
62. Chen SG, Leu YL, Cheng ML, Ting SC, Liu CC, Wang SD, Yang CH, Hung CY, Sakurai H, Chen KH, Ho HY. 2017. Anti-enterovirus 71 activities of Melissa officinalis extract and its biologically active constituent rosmarinic acid. Scientific reports 7:12264.
63. Ho HY, Cheng ML, Weng SF, Leu YL, Chiu DT. 2009. Antiviral effect of epigallocatechin gallate on enterovirus 71. Journal of Agricultural and Food Chemistry 57:6140-6147.
64. Chen SG, Cheng ML, Chen KH, Horng JT, Liu CC, Wang SM, Sakurai H, Leu YL, Wang SD, Ho HY. 2017. Antiviral activities of Schizonepeta tenuifolia Briq. against enterovirus 71 in vitro and in vivo. Scientific reports 7:935.
65. Liang CT, Chueh LL, Pang VF, Zhuo YX, Liang SC, Yu CK, Chiang H, Lee CC, Liu CH. 2007. A non-biotin polymerized horseradish-peroxidase method for the immunohistochemical diagnosis of canine distemper. Journal of Comparative Pathology 136:57-64.
66. Lee JK, Tansey MG. 2013. Microglia isolation from adult mouse brain. Methods in Molecular Biology 1041:17-23.
67. Dionne KR, Tyler KL. 2013. Slice culture modeling of central nervous system (CNS) viral infection. Methods in Molecular Biology 1078:97-117.
68. Wang SM, Lei HY, Liu CC. 2012. Cytokine immunopathogenesis of enterovirus 71 brain stem encephalitis. Clinical & Developmental Immunology 2012:876241.
69. Graeber MB, Li W, Rodriguez ML. 2011. Role of microglia in CNS inflammation. FEBS Letters 585:3798-3805.
70. Imai Y, Kohsaka S. 2002. Intracellular signaling in M-CSF-induced microglia activation: role of Iba1. Glia 40:164-174.
71. Franco R, Fernandez-Suarez D. 2015. Alternatively activated microglia and macrophages in the central nervous system. Progress in neurobiology 131:65-86.
72. Conrady CD, Zheng M, van Rooijen N, Drevets DA, Royer D, Alleman A, Carr DJ. 2013. Microglia and a functional type I IFN pathway are required to counter HSV-1-driven brain lateral ventricle enlargement and encephalitis. Journal of Immunology 190:2807-2817.
73. Chhor V, Le Charpentier T, Lebon S, Ore MV, Celador IL, Josserand J, Degos V, Jacotot E, Hagberg H, Savman K, Mallard C, Gressens P, Fleiss B. 2013. Characterization of phenotype markers and neuronotoxic potential of polarised primary microglia in vitro. Brain, Behavior, and Immunity 32:70-85.
74. Fu X, Zunich SM, O'Connor JC, Kavelaars A, Dantzer R, Kelley KW. 2010. Central administration of lipopolysaccharide induces depressive-like behavior in vivo and activates brain indoleamine 2,3 dioxygenase in murine organotypic hippocampal slice cultures. Journal of Neuroinflammation 7:43.
75. Zhang Y, Cui W, Liu L, Wang J, Zhao H, Liao Y, Na R, Dong C, Wang L, Xie Z, Gao J, Cui P, Zhang X, Li Q. 2011. Pathogenesis study of enterovirus 71 infection in rhesus monkeys. Laboratory investigation 91:1337-1350.
76. Drummond CG, Bolock AM, Ma C, Luke CJ, Good M, Coyne CB. 2017. Enteroviruses infect human enteroids and induce antiviral signaling in a cell lineage-specific manner. Proceedings of the National Academy of Sciences of the United States of America 114:1672-1677.
77. Chen CS, Yao YC, Lin SC, Lee YP, Wang YF, Wang JR, Liu CC, Lei HY, Yu CK. 2007. Retrograde axonal transport: a major transmission route of enterovirus 71 in mice. Journal of virology 81:8996-9003.
78. Tan SH, Ong KC, Wong KT. 2014. Enterovirus 71 can directly infect the brainstem via cranial nerves and infection can be ameliorated by passive immunization. Journal of neuropathology and experimental neurology 73:999-1008.
79. Lin P, Gao L, Huang Y, Chen Q, Shen H. 2015. An enterovirus 71 strain causes skeletal muscle damage in infected mice. International journal of clinical and experimental pathology 8:3460-3468.
80. Liou AT, Wu SY, Liao CC, Chang YS, Chang CS, Shih C. 2016. A new animal model containing human SCARB2 and lacking stat-1 is highly susceptible to EV71. Scientific reports 6:31151.
81. Lin JY, Shih SR. 2014. Cell and tissue tropism of enterovirus 71 and other enteroviruses infections. Journal of biomedical science 21:18.
82. Robinson CM, Pfeiffer JK. 2014. Viruses and the Microbiota. Annual review of virology 1:55-69.
83. Ooi MH, Wong SC, Lewthwaite P, Cardosa MJ, Solomon T. 2010. Clinical features, diagnosis, and management of enterovirus 71. The Lancet Neurology 9:1097-1105.
84. Tourneur E, Chassin C. 2013. Neonatal immune adaptation of the gut and its role during infections. Clinical & Developmental Immunology 2013:270301.
85. Dzidic M, Boix-Amoros A, Selma-Royo M, Mira A, Collado MC. 2018. Gut Microbiota and Mucosal Immunity in the Neonate. Medical sciences 6.
86. Goto K, Sanefuji M, Kusuhara K, Nishimura Y, Shimizu H, Kira R, Torisu H, Hara T. 2009. Rhombencephalitis and coxsackievirus A16. Emerging Infectious Diseases 15:1689-1691.
87. Wright HT, Jr., Landing BH, Lennette EH, Mc AR. 1963. Fatal infection in an infant associated with Coxsackie virus group A, type 16. The New England journal of medicine 268:1041-1044.
88. Xu W, Liu CF, Yan L, Li JJ, Wang LJ, Qi Y, Cheng RB, Xiong XY. 2012. Distribution of enteroviruses in hospitalized children with hand, foot and mouth disease and relationship between pathogens and nervous system complications. Virology journal 9:8.
89. Kettenmann H, Hanisch UK, Noda M, Verkhratsky A. 2011. Physiology of microglia. Physiological reviews 91:461-553.
90. Hurgin V, Novick D, Werman A, Dinarello CA, Rubinstein M. 2007. Antiviral and immunoregulatory activities of IFN-gamma depend on constitutively expressed IL-1alpha. Proceedings of the National Academy of Sciences of the United States of America 104:5044-5049.
91. Liddelow SA, Barres BA. 2017. Reactive Astrocytes: Production, Function, and Therapeutic Potential. Immunity 46:957-967.
92. Liddelow SA, Guttenplan KA, Clarke LE, Bennett FC, Bohlen CJ, Schirmer L, Bennett ML, Munch AE, Chung WS, Peterson TC, Wilton DK, Frouin A, Napier BA, Panicker N, Kumar M, Buckwalter MS, Rowitch DH, Dawson VL, Dawson TM, Stevens B, Barres BA. 2017. Neurotoxic reactive astrocytes are induced by activated microglia. Nature 541:481-487.
93. Sofroniew MV, Vinters HV. 2010. Astrocytes: biology and pathology. Acta neuropathologica 119:7-35.
94. Spandidos A, Wang X, Wang H, Seed B. 2010. PrimerBank: a resource of human and mouse PCR primer pairs for gene expression detection and quantification. Nucleic acids research 38:D792-799.
95. Vinet J, Weering HR, Heinrich A, Kalin RE, Wegner A, Brouwer N, Heppner FL, Rooijen N, Boddeke HW, Biber K. 2012. Neuroprotective function for ramified microglia in hippocampal excitotoxicity. Journal of neuroinflammation 9:27.