Increased and Prolonged Human Norovirus Infection in RAG2/IL2RG Deficient Gnotobiotic Pigs with Severe Combined Immunodeficiency

Overview

Journal Sci Rep

Specialty Science

Date 2016 Apr 28

PMID 27118081

Citations 54

Authors

Shaohua Lei

Junghyun Ryu

Ke Wen

Erica Twitchell

Tammy Bui

Ashwin Ramesh

Mariah Weiss

Guohua Li

Helen Samuel

Sherrie Clark-Deener

Xi Jiang

Kiho Lee

Lijuan Yuan

Affiliations

Soon will be listed here.

Abstract

Application of genetically engineered (GE) large animals carrying multi-allelic modifications has been hampered by low efficiency in production and extended gestation period compared to rodents. Here, we rapidly generated RAG2/IL2RG double knockout pigs using direct injection of CRISPR/Cas9 system into developing embryos. RAG2/IL2RG deficient pigs were immunodeficient, characterized by depletion of lymphocytes and either absence of or structurally abnormal immune organs. Pigs were maintained in gnotobiotic facility and evaluated for human norovirus (HuNoV) infection. HuNoV shedding lasted for 16 days in wild type pigs, compared to 27 days (until the end of trials) in RAG2/IL2RG deficient pigs. Additionally, higher HuNoV titers were detected in intestinal tissues and contents and in blood, indicating increased and prolonged HuNoV infection in RAG2/IL2RG deficient pigs and the importance of lymphocytes in HuNoV clearance. These results suggest that GE immunodeficient gnotobiotic pigs serve as a novel model for biomedical research and will facilitate HuNoV studies.

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References

Mai H, Gao Y, Cong X, Wang H, Liu N, Huang X . GII.4 Sydney_2012 norovirus infection in immunocompromised patients in Beijing and its rapid evolution in vivo. J Med Virol. 2015; 88(2):224-33. DOI: 10.1002/jmv.24332. View

Karve S, Krishnarajah G, Korsnes J, Cassidy A, Candrilli S . Burden of acute gastroenteritis, norovirus and rotavirus in a managed care population. Hum Vaccin Immunother. 2014; 10(6):1544-56. PMC: 5396247. DOI: 10.4161/hv.28704. View

Zhu S, Regev D, Watanabe M, Hickman D, Moussatche N, Jesus D . Identification of immune and viral correlates of norovirus protective immunity through comparative study of intra-cluster norovirus strains. PLoS Pathog. 2013; 9(9):e1003592. PMC: 3764223. DOI: 10.1371/journal.ppat.1003592. View

Lai L, Kolber-Simonds D, Park K, Cheong H, Greenstein J, Im G . Production of alpha-1,3-galactosyltransferase knockout pigs by nuclear transfer cloning. Science. 2002; 295(5557):1089-92. DOI: 10.1126/science.1068228. View

Bui T, Kocher J, Li Y, Wen K, Li G, Liu F . Median infectious dose of human norovirus GII.4 in gnotobiotic pigs is decreased by simvastatin treatment and increased by age. J Gen Virol. 2013; 94(Pt 9):2005-2016. PMC: 3749057. DOI: 10.1099/vir.0.054080-0. View

Jung K, Wang Q, Kim Y, Scheuer K, Zhang Z, Shen Q . The effects of simvastatin or interferon-α on infectivity of human norovirus using a gnotobiotic pig model for the study of antivirals. PLoS One. 2012; 7(7):e41619. PMC: 3402445. DOI: 10.1371/journal.pone.0041619. View

Whitworth K, Lee K, Benne J, Beaton B, Spate L, Murphy S . Use of the CRISPR/Cas9 system to produce genetically engineered pigs from in vitro-derived oocytes and embryos. Biol Reprod. 2014; 91(3):78. PMC: 4435063. DOI: 10.1095/biolreprod.114.121723. View

Carter D, Lai L, Park K, Samuel M, Lattimer J, Jordan K . Phenotyping of transgenic cloned piglets. Cloning Stem Cells. 2002; 4(2):131-45. DOI: 10.1089/153623002320253319. View

Karst S . Pathogenesis of noroviruses, emerging RNA viruses. Viruses. 2011; 2(3):748-781. PMC: 3185648. DOI: 10.3390/v2030748. View

10.

Yoshioka K, Suzuki C, Tanaka A, Iwamura S . Birth of piglets derived from porcine zygotes cultured in a chemically defined medium. Biol Reprod. 2001; 66(1):112-9. DOI: 10.1095/biolreprod66.1.112. View

11.

Takanashi S, Hashira S, Matsunaga T, Yoshida A, Shiota T, Tung P . Detection, genetic characterization, and quantification of norovirus RNA from sera of children with gastroenteritis. J Clin Virol. 2009; 44(2):161-3. DOI: 10.1016/j.jcv.2008.11.011. View

12.

Kaufman S, Green K, Korba B . Treatment of norovirus infections: moving antivirals from the bench to the bedside. Antiviral Res. 2014; 105:80-91. PMC: 4793406. DOI: 10.1016/j.antiviral.2014.02.012. View

13.

Patel M, Widdowson M, Glass R, Akazawa K, Vinje J, Parashar U . Systematic literature review of role of noroviruses in sporadic gastroenteritis. Emerg Infect Dis. 2008; 14(8):1224-31. PMC: 2600393. DOI: 10.3201/eid1408.071114. View

14.

Kageyama T, Kojima S, Shinohara M, Uchida K, Fukushi S, Hoshino F . Broadly reactive and highly sensitive assay for Norwalk-like viruses based on real-time quantitative reverse transcription-PCR. J Clin Microbiol. 2003; 41(4):1548-57. PMC: 153860. DOI: 10.1128/JCM.41.4.1548-1557.2003. View

15.

Wang X, Zhou J, Cao C, Huang J, Hai T, Wang Y . Efficient CRISPR/Cas9-mediated biallelic gene disruption and site-specific knockin after rapid selection of highly active sgRNAs in pigs. Sci Rep. 2015; 5:13348. PMC: 4543986. DOI: 10.1038/srep13348. View

16.

Gastanaduy P, Hall A, Curns A, Parashar U, Lopman B . Burden of norovirus gastroenteritis in the ambulatory setting--United States, 2001-2009. J Infect Dis. 2013; 207(7):1058-65. DOI: 10.1093/infdis/jis942. View

17.

Tan M, Jiang X . Vaccine against norovirus. Hum Vaccin Immunother. 2014; 10(6):1449-56. PMC: 5396253. DOI: 10.4161/hv.28626. View

18.

Yuan L, Ward L, Rosen B, To T, Saif L . Systematic and intestinal antibody-secreting cell responses and correlates of protective immunity to human rotavirus in a gnotobiotic pig model of disease. J Virol. 1996; 70(5):3075-83. PMC: 190169. DOI: 10.1128/JVI.70.5.3075-3083.1996. View

19.

Kocher J, Bui T, Giri-Rachman E, Wen K, Li G, Yang X . Intranasal P particle vaccine provided partial cross-variant protection against human GII.4 norovirus diarrhea in gnotobiotic pigs. J Virol. 2014; 88(17):9728-43. PMC: 4136312. DOI: 10.1128/JVI.01249-14. View

20.

Whitworth K, Rowland R, Ewen C, Trible B, Kerrigan M, Cino-Ozuna A . Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus. Nat Biotechnol. 2015; 34(1):20-2. DOI: 10.1038/nbt.3434. View