建立表达犬NECTIN4的牛源细胞株用于培养犬瘟热病毒
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摘要
NECTIN4是犬瘟热病毒的受体之一。通过在牛肾MDBK细胞株中表达NECTIN4,建立MDBK-N4细胞株用于培养犬瘟热病毒。接种犬瘟热病毒后,MDBK-N4细胞能够扩增病毒并产生细胞病变,这说明病毒受体的表达增强了病毒与细胞膜表面的结合能力,进而改变细胞对病毒的敏感性。这一发现为工业疫苗生产细胞株的开发提供了新的思路。
Nectin cell adhesion molecule 4(NECTIN4) is one of the receptors for canine distemper virus(CDV). In this study, a Madin-Darby Bovine Kidney(MDBK) cell line expressing canine NECTIN4, designated MDBK-N4, was established for CDV culture. We found that MDBK-N4 cell line could multiply the virus and it was caused typical cytopathic effect(CPE) after CDV inoculation, which demonstrated that transfection with viral receptor complementary DNA may enables a non-susceptible cell line to bind virus, support virus replication and develop CPE. Viral receptors expression provides a new way to develop a cell line for vaccine production in industry.
Nectin cell adhesion molecule 4(NECTIN4) is one of the receptors for canine distemper virus(CDV). In this study, a Madin-Darby Bovine Kidney(MDBK) cell line expressing canine NECTIN4, designated MDBK-N4, was established for CDV culture. We found that MDBK-N4 cell line could multiply the virus and it was caused typical cytopathic effect(CPE) after CDV inoculation, which demonstrated that transfection with viral receptor complementary DNA may enables a non-susceptible cell line to bind virus, support virus replication and develop CPE. Viral receptors expression provides a new way to develop a cell line for vaccine production in industry.
引文
[1]LOOTS A K, MITCHELL E, DALTON D L, et al. Advances in canine distemper virus pathogenesis research: a wildlife perspective [J]. The Journal of General Virology, 2017, 98(3): 311-321.
[2]MARTINEZ-GUTIERREZ M, RUIZ-SAENZ J. Diversity of susceptible hosts in canine distemper virus infection: a systematic review and data synthesis [J]. BMC Veterinary Research, 2016, 12(1): 1-11.
[3]王雅文, 赵建军, 闫喜军. 犬瘟热病毒受体及其跨宿主感染 [J]. 病毒学报, 2017, 33(3): 477-482.
[4]ATHANASIOU L V, KANTERE M C, KYRIAKIS C S, et al. Evaluation of a direct immunofluorescent assay and/or conjunctival cytology for detection of canine distemper virus antigen [J]. Viral Immunology, 2018, 31(3):272-275.
[5]SANTIAGO C, CELMA M L, STEHLE T, et al. Structure of the measles virus hemagglutinin bound to the CD46 receptor [J]. Nature Structural & Molecular Biology, 2010, 17(1): 124-129.
[6]HASHIGUCHI T, OSE T, KUBOTA M, et al. Structure of the measles virus hemagglutinin bound to its cellular receptor SLAM [J]. Nature Structural & Molecular Biology, 2011, 18(2): 135-141.
[7]DELPEUT S, NOYCE R S, RICHARDSON C D. The tumor-associated marker, PVRL4 (NECTIN4), is the epithelial receptor for morbilliviruses [J]. Viruses, 2014, 6(6): 2268-2286.
[8]DELPEUT S, NOYCE R S, RICHARDSON C D. The V domain of dog PVRL4 (NECTIN4) mediates canine distemper virus entry and virus cell-to-cell spread [J]. Virology, 2014, 454-455(4): 109-117.
[9]LIN L T, RICHARDSON C D. The host cell receptors for measles virus and their interaction with the viral hemagglutinin (H) protein [J]. Viruses, 2016, 8(9): 250.
[10]ELGENDY R, GIANTIN M, DACASTO M. Transcriptomic characterization of bovine primary cultured hepatocytes; a cross-comparison with a bovine liver and the Madin-Darby bovine kidney cells [J]. Research in Veterinary Science, 2017, 113(1): 40-49.
[11]SAMANTA D, ALMO S C. Nectin family of cell-adhesion molecules: structural and molecular aspects of function and specificity [J]. Cellular and Molecular Life Sciences : CMLS, 2015, 72(4): 645-658.
[12]NOYCE R S, BONDRE D G, HA M N, et al. Tumor cell marker PVRL4 (NECTIN4) is an epithelial cell receptor for measles virus [J]. PLoS Pathogens, 2011, 7(8): e1002240.
[13]FENG N, LIU Y X, WANG J Z, et al. Canine distemper virus isolated from a monkey efficiently replicates on Vero cells expressing non-human primate SLAM receptors but not human SLAM receptor [J]. BMC Veterinary Research, 2016, 12(160): 7.
[14]PRATAKPIRIYA W, PING TEH A P, RADTANAKATIKANON A, et al. Expression of canine distemper virus receptor NECTIN4 in the central nervous system of dogs [J]. Scientific Reports, 2017, 7(1): 349.
[15]张薇, 闫琰, 白鲁根, 等. 稳定表达犬转铁蛋白受体CHO细胞系的建立和鉴定 [J]. 四川大学学报(医学版), 2017, 48(1): 57-60.
[16]ZHANG J, LIU W, CHEN W, et al. Development of an immunoperoxidase monolayer assay for the detection of antibodies against peste des petits ruminants virus based on BHK-21 cell line stably expressing the goat signaling lymphocyte activation molecule [J]. PLoS One, 2016, 11(10): e0165088.
[17]TAPIA F, VAZQUEZ-RAMIREZ D, GENZEL Y, et al. Bioreactors for high cell density and continuous multi-stage cultivations: options for process intensification in cell culture-based viral vaccine production [J]. Applied Microbiology and Biotechnology, 2016, 100(5): 2121-2132.
[18]杨艳坤, 郭雯雯, 隋涌, 等. 规模化哺乳细胞培养技术生产病毒疫苗——过程工程和生物反应器 [J]. 生物产业技术, 2015, 1(1): 7-16.
[19]XU N, MA C, OU J, et al. Comparative proteomic analysis of three Chinese hamster ovary (CHO) host cells [J]. Biochemical Engineering Journal, 2017, 124(1): 122-129.
[20]GENZEL Y. Designing cell lines for viral vaccine production: Where do we stand? [J]. Biotechnology Journal, 2015, 10(5): 728-740.
[2]MARTINEZ-GUTIERREZ M, RUIZ-SAENZ J. Diversity of susceptible hosts in canine distemper virus infection: a systematic review and data synthesis [J]. BMC Veterinary Research, 2016, 12(1): 1-11.
[3]王雅文, 赵建军, 闫喜军. 犬瘟热病毒受体及其跨宿主感染 [J]. 病毒学报, 2017, 33(3): 477-482.
[4]ATHANASIOU L V, KANTERE M C, KYRIAKIS C S, et al. Evaluation of a direct immunofluorescent assay and/or conjunctival cytology for detection of canine distemper virus antigen [J]. Viral Immunology, 2018, 31(3):272-275.
[5]SANTIAGO C, CELMA M L, STEHLE T, et al. Structure of the measles virus hemagglutinin bound to the CD46 receptor [J]. Nature Structural & Molecular Biology, 2010, 17(1): 124-129.
[6]HASHIGUCHI T, OSE T, KUBOTA M, et al. Structure of the measles virus hemagglutinin bound to its cellular receptor SLAM [J]. Nature Structural & Molecular Biology, 2011, 18(2): 135-141.
[7]DELPEUT S, NOYCE R S, RICHARDSON C D. The tumor-associated marker, PVRL4 (NECTIN4), is the epithelial receptor for morbilliviruses [J]. Viruses, 2014, 6(6): 2268-2286.
[8]DELPEUT S, NOYCE R S, RICHARDSON C D. The V domain of dog PVRL4 (NECTIN4) mediates canine distemper virus entry and virus cell-to-cell spread [J]. Virology, 2014, 454-455(4): 109-117.
[9]LIN L T, RICHARDSON C D. The host cell receptors for measles virus and their interaction with the viral hemagglutinin (H) protein [J]. Viruses, 2016, 8(9): 250.
[10]ELGENDY R, GIANTIN M, DACASTO M. Transcriptomic characterization of bovine primary cultured hepatocytes; a cross-comparison with a bovine liver and the Madin-Darby bovine kidney cells [J]. Research in Veterinary Science, 2017, 113(1): 40-49.
[11]SAMANTA D, ALMO S C. Nectin family of cell-adhesion molecules: structural and molecular aspects of function and specificity [J]. Cellular and Molecular Life Sciences : CMLS, 2015, 72(4): 645-658.
[12]NOYCE R S, BONDRE D G, HA M N, et al. Tumor cell marker PVRL4 (NECTIN4) is an epithelial cell receptor for measles virus [J]. PLoS Pathogens, 2011, 7(8): e1002240.
[13]FENG N, LIU Y X, WANG J Z, et al. Canine distemper virus isolated from a monkey efficiently replicates on Vero cells expressing non-human primate SLAM receptors but not human SLAM receptor [J]. BMC Veterinary Research, 2016, 12(160): 7.
[14]PRATAKPIRIYA W, PING TEH A P, RADTANAKATIKANON A, et al. Expression of canine distemper virus receptor NECTIN4 in the central nervous system of dogs [J]. Scientific Reports, 2017, 7(1): 349.
[15]张薇, 闫琰, 白鲁根, 等. 稳定表达犬转铁蛋白受体CHO细胞系的建立和鉴定 [J]. 四川大学学报(医学版), 2017, 48(1): 57-60.
[16]ZHANG J, LIU W, CHEN W, et al. Development of an immunoperoxidase monolayer assay for the detection of antibodies against peste des petits ruminants virus based on BHK-21 cell line stably expressing the goat signaling lymphocyte activation molecule [J]. PLoS One, 2016, 11(10): e0165088.
[17]TAPIA F, VAZQUEZ-RAMIREZ D, GENZEL Y, et al. Bioreactors for high cell density and continuous multi-stage cultivations: options for process intensification in cell culture-based viral vaccine production [J]. Applied Microbiology and Biotechnology, 2016, 100(5): 2121-2132.
[18]杨艳坤, 郭雯雯, 隋涌, 等. 规模化哺乳细胞培养技术生产病毒疫苗——过程工程和生物反应器 [J]. 生物产业技术, 2015, 1(1): 7-16.
[19]XU N, MA C, OU J, et al. Comparative proteomic analysis of three Chinese hamster ovary (CHO) host cells [J]. Biochemical Engineering Journal, 2017, 124(1): 122-129.
[20]GENZEL Y. Designing cell lines for viral vaccine production: Where do we stand? [J]. Biotechnology Journal, 2015, 10(5): 728-740.