ZHANG Jian, WU Zhenfang, YANG Huaqiang. Resistance to blue ear disease and production performance assessment of CD163 gene-edited Large White pigs[J]. Journal of South China Agricultural University, 2023, 44(3): 333-339. DOI: 10.7671/j.issn.1001-411X.202203033
    Citation: ZHANG Jian, WU Zhenfang, YANG Huaqiang. Resistance to blue ear disease and production performance assessment of CD163 gene-edited Large White pigs[J]. Journal of South China Agricultural University, 2023, 44(3): 333-339. DOI: 10.7671/j.issn.1001-411X.202203033

    Resistance to blue ear disease and production performance assessment of CD163 gene-edited Large White pigs

    More Information
    • Received Date: March 17, 2022
    • Available Online: May 17, 2023
    • Objective 

      The purpose of this study was to generate CD163 gene knockout (CD163-KO) Large White pigs by CRISPR/Cas9 gene editing and somatic cell nuclear transfer technologies, investigate the resistance to blue ear disease and the biosafety effect including physiology, productive and reproductive performances of the gene knockout pigs, and assess the main production performances of CD163-KO Large White pigs.

      Method 

      In this study, the 11 CD163-KO pigs and five age- and body weight-matched wild type Large White pigs were challenged with NADC30-like strain of porcine reproductive and respiratory syndrome virus (PRRSV). The rectal temperature, PRRSV antibody and virus variation were monitored continuously for 14 days. The lung tissues were examined by immunofluorescence of PRRSV antigen. Expression of CD163 protein on the surface of pulmonary alveolar macrophages in wild type and CD163-KO Large White pigs were examined through immunofluorescence staining. We compared the differentiation potential of monocytes into macrophages between CD163-KO and wild type pigs, and observed their uptake capacities to hemoglobin-haptoglobin complex. In addition, we analyzed the growth and reproductive production of the boars between CD163-KO pigs and wild type control to assess their biosafety and breeding value.

      Result 

      CD163-KO pigs were completely resistant to NADC30-like strain without impairing the biological function associated with the modified gene, as well as productive and reproductive performances.

      Conclusion 

      This study is an evidence and supplement of CD163-KO pigs resistance to blue ear disease, and demonstrates that CD163 gene knockout has no potentially negative effects on production performance, which provides evidences for the biosecurity of CD163-KO pigs.

    • [1]
      HAN J, ZHOU L, GE X, et al. Pathogenesis and control of the Chinese highly pathogenic porcine reproductive and respiratory syndrome virus[J]. Veterinary Microbiology, 2017, 209: 30-47. doi: 10.1016/j.vetmic.2017.02.020
      [2]
      TIAN K, YU X, ZHAO T, et al. Emergence of fatal PRRSV variants: Unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique hallmark[J]. PLoS One, 2007, 2(6): e526. doi: 10.1371/journal.pone.0000526
      [3]
      ZHAO K, YE C, CHANG X, et al. Importation and recombination are responsible for the latest emergence of highly pathogenic porcine reproductive and respiratory syndrome virus in China[J]. Journal of Virology, 2015, 89(20): 10712-10716. doi: 10.1128/JVI.01446-15
      [4]
      WANG H M, LIU Y G, TANG Y D, et al. A natural recombinant PRRSV between HP-PRRSV JXA1-like and NADC30-like strains[J]. Transboundary and Emerging Diseases, 2018, 65(4): 1078-1086. doi: 10.1111/tbed.12852
      [5]
      YU Y, ZHANG Q, CAO Z, et al. Recent advances in porcine reproductive and respiratory syndrome virus NADC30-like research in China: Molecular characterization, pathogenicity, and control[J]. Frontiers in Microbiology, 2022, 12: 791313. doi: 10.3389/fmicb.2021.791313.
      [6]
      ZHOU L, YANG B, XU L, et al. Efficacy evaluation of three modified-live virus vaccines against a strain of porcine reproductive and respiratory syndrome virus NADC30-like[J]. Veterinary Microbiology, 2017, 207: 108-116. doi: 10.1016/j.vetmic.2017.05.031
      [7]
      CHAI W, LIU Z, SUN Z, et al. Efficacy of two porcine reproductive and respiratory syndrome (PRRS) modified-live virus (MLV) vaccines against heterologous NADC30-like PRRS virus challenge[J]. Veterinary Microbiology, 2020, 248: 108805. doi: 10.1016/j.vetmic.2020.108805.
      [8]
      RENUKARADHYA G J, MENG X J, CALVERT J G, et al. Inactivated and subunit vaccines against porcine reproductive and respiratory syndrome: Current status and future direction[J]. Vaccine, 2015, 33(27): 3065-3072. doi: 10.1016/j.vaccine.2015.04.102
      [9]
      熊胜利, 龙清孟, 陈大方, 等. 抗病育种技术在引进美国SPF种猪后代选育中应用探究[J]. 养猪, 2014(3): 73-74. doi: 10.13257/j.cnki.21-1104/s.2014.03.028
      [10]
      YUAN H, YANG L, ZHANG Y, et al. Current status of genetically modified pigs that are resistant to virus infection[J]. Viruses, 2022, 14(2): 417. doi: 10.3390/v14020417.
      [11]
      PROUDFOOT C, LILLICO S, TAIT-BURKARD C. Genome editing for disease resistance in pigs and chickens[J]. Animal Frontiers, 2019, 9(3): 6-12. doi: 10.1093/af/vfz013
      [12]
      WHITWORTH K M, ROWLAND R R R, EWEN C L, et al. Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus[J]. Nature Biotechnology, 2016, 34(1): 20-22. doi: 10.1038/nbt.3434
      [13]
      YANG H, ZHANG J, ZHANG X, et al. CD163 knockout pigs are fully resistant to highly pathogenic porcine reproductive and respiratory syndrome virus[J]. Antiviral Research, 2018, 151: 63-70.
      [14]
      BURKARD C, LILLICO S G, REID E, et al. Precision engineering for PRRSV resistance in pigs: Macrophages from genome edited pigs lacking CD163 SRCR5 domain are fully resistant to both PRRSV genotypes while maintaining biological function[J]. PLOS Pathogens, 2017, 13(2): e1006206. doi: 10.1371/journal.ppat.1006206
      [15]
      WELCH S K W, CALVERT J G. A brief review of CD163 and its role in PRRSV infection[J]. Virus Research, 2010, 154(1/2): 98-103. doi: 10.1016/j.virusres.2010.07.018
      [16]
      XU Y, WU S, LI Y, et al. A porcine alveolar macrophage cell line stably expressing CD163 demonstrates virus replication and cytokine secretion characteristics similar to primary alveolar macrophages following PRRSV infection[J]. Veterinary Microbiology, 2020, 244: 108690. doi: 10.1016/j.vetmic.2020.108690.
      [17]
      XU K, ZHOU Y, MU Y, et al. CD163 and pAPN double-knockout pigs are resistant to PRRSV and TGEV and exhibit decreased susceptibility to PDCoV while maintaining normal production performance[J]. eLife, 2020, 9: e57132.
      [18]
      韩晓松, 高杨, 刘海龙, 等. 利用CRISPR/Cas9技术制备CD163基因SRCR5序列敲除猪[J]. 农业生物技术学报, 2020, 28(9): 1535-1542.
      [19]
      GUO C, WANG M, ZHU Z, et al. Highly efficient generation of pigs harboring a partial deletion of the CD163 SRCR5 domain, which are fully resistant to porcine reproductive and respiratory syndrome virus 2 infection[J]. Frontiers in Immunology, 2019, 10: 1846. doi: 10.3389/fimmu.2019.01846.
      [20]
      CHEN J, WANG H, BAI J, et al. Generation of pigs resistant to highly pathogenic-porcine reproductive and respiratory syndrome virus through gene editing of CD163[J]. International Journal of Biological Sciences, 2019, 15(2): 481-492. doi: 10.7150/ijbs.25862
      [21]
      VAN GORP H, VAN BREEDAM W, DELPUTTE P L, et al. Sialoadhesin and CD163 join forces during entry of the porcine reproductive and respiratory syndrome virus[J]. Journal of General Virology, 2008, 89: 2943-2953. doi: 10.1099/vir.0.2008/005009-0
      [22]
      ZHANG Q, YOO D. PRRS virus receptors and their role for pathogenesis[J]. Veterinary Microbiology, 2015, 177(3/4): 229-241.
      [23]
      VAN BREEDAM W, VERBEECK M, CHRISTIAENS I, et al. Porcine, murine and human sialoadhesin (Sn/Siglec-1/CD169): Portals for porcine reproductive and respiratory syndrome virus entry into target cells[J]. Journal of General Virology, 2013, 94: 1955-1960.
      [24]
      GRAVERSEN J H, MADSEN M, MOESTRUP S K. CD163: A signal receptor scavenging haptoglobin-hemoglobin complexes from plasma[J]. International Journal of Biochemistry & Cell Biology, 2002, 34(4): 309-314.
      [25]
      KRISTIANSEN M, GRAVERSEN J H, JACOBSEN C, et al. Identification of the haemoglobin scavenger receptor[J]. Nature, 2001, 409(6817): 198-201. doi: 10.1038/35051594
      [26]
      SCHAER C A, VALLELIAN F, IMHOF A, et al. CD163-expressing monocytes constitute an endotoxin-sensitive Hb clearance compartment within the vascular system[J]. Journal of Leukocyte Biology, 2007, 82(1): 106-110. doi: 10.1189/jlb.0706453
      [27]
      石俊松, 周荣, 曾海玉, 等. 体细胞克隆猪繁殖性能及后代生长性能评估[J]. 华南农业大学学报, 2019, 40(S1): 100-103.
      [28]
      ADACHI N, YAMAGUCHI D, WATANABE A, et al. Growth, reproductive performance, carcass characteristics and meat quality in F1 and F2 progenies of somatic cell-cloned pigs[J]. Joural of Reproduction and Development, 2014, 60(2): 100-105. doi: 10.1262/jrd.2012-167
      [29]
      KAWARASAKI T, ENYA S, OTAKE M, et al. Reproductive performance and expression of imprinted genes in somatic cell cloned boars[J]. Animal Science Journal, 2017, 88(11): 1801-1810. doi: 10.1111/asj.12838
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