Abstract：Cluster of differentiation 163(CD163) protein is a critical receptor for the infection of Porcine reproductive and respiratory syndrome virus(PRRSV). In recent years, gene-editing technology has been utilized to breed PRRSV-resistant pigs. This review systematically examines the structure and physiological functions of the CD163 protein, summarizes domestic and international research cases in which PRRSV-resistant pigs have been successfully developed through various strategies(including complete deletion, partial deletion, replacement of the cysteine-rich scavenger receptor 5(SRCR5) domain, and single-base editing), and focuses on evaluating the impact of these genetic modifications on other key physiological functions of CD163, such as clearance of hemoglobin-haptoglobin complexes, promotion of erythrocyte development, and immune regulation, in order to provide scientific evidence and references for the safety of commercialized gene-edited disease-resistant animals. Substantial experimental evidence reveals that CD163-edited pigs exhibit complete resistance to PRRSV infection while retaining all normal physiological functions, including the clearance of free hemoglobin in plasma, the promotion of erythroblast development, and its function as a receptor for the TNF-like weak inducer of apoptosis(TWEAK). These findings provide crucial safety assurances for the commercial application of PRRSV-resistant gene-edited swine.
刘夏仪, 汤波, 李宁. 猪CD163基因编辑对PRRSV抗性及其他生理功能影响的分析[J]. 农业生物技术学报, 2026, 34(8): 1792-1802.
LIU Xia-Yi, TANG Bo, LI Ning. Analysis of the Impact of Porcine(Sus scrofa) CD163 Gene Editing on PRRSV Resistance and Other Physiological Functions. 农业生物技术学报, 2026, 34(8): 1792-1802.
[1] 韩晓松, 高杨, 刘海龙, 等. 2020. 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, 28(09): 1535-1542. (Han X S, Gao Y, Liu H L, et al.2020. Generation of CD163 gene SRCR5 deleted pig(Sus scrofa) via CRISPR/Cas9[J]. Journal of Agricultural Biotechnology, 28(09): 1535-1542.) [2] 侯亚芝, 张晋欣, 陈小飞. 2022. 可溶性血红蛋白清道夫受体163在不同疾病中的研究进展[J]. 中西医结合心脑血管病杂志, 20(18): 3341-3348. (Hou Y Z, Zhang J X, Chen X F.2022. Research progress on sCD163 in various pathological conditions[J]. Chinese Journal of Integrative Medicine on Cardio/Cerebrovascular Disease, 20(18): 3341-3348.) [3] 李海洲, 夏豪, 王薇. 2007. 游离血红蛋白及其受体CD163在大鼠动脉粥样硬化过程中的作用[J]. 中国病理生理杂志, 23(9): 1695-1699. (Li H Z, Xia H, Wang W.2007. Role of free hemoglobin and its receptor CD163 in the rat atherosclerosis formation[J]. Chinese Journal of Pathophysiology, 23(9): 1695-1699.) [4] 刘玉君. 2022. CD163基因rs4883263和rs11054072位点多态性与冠心病患者血脂水平以及阿托伐他汀降脂效果的相关性研究[D]. 硕士学位论文, 青岛大学, 导师: 隋忠国, pp. 44-49. (Liu Y J.2022. The relationship between CD163 rs4883263 and rs11054072 gene polymorphisms and blood lipid level in patients with coronary heart disease and the lipid-lowering effect of atorvastatin[D]. Thesis for M.S., Qingdao University, Supervisor: Sui Z G, pp. 44-49.) [5] 梅万春. 2021. CD163对小鼠缺血性心肌病心室重塑的作用研究[D]. 硕士学位论文, 厦门大学, 导师: 戴翠莲: (Mei W C. 2021. The role of CD163 inventricular remodeling in mice with ischemic cardiomyopathy[D].Thesis for M.S., Xiamen University, Supervisor: Dai C L, pp. 32-41.) [6] 任吉龙. 2023. 应用Cas12iMax进行多基因编辑提高家畜肌肉产量及抗病能力的研究[D]. 博士学位论文, 中国农业科学院, 导师: 朱化彬, pp. 36-60. (Ren J L.2023. Improve meat production and virus resistance by simultaneously editing multiple genes in livestock using Cas12iMax[D]. Thesis for Ph.D., Chinese Academy of Agricultural Sciences, Supervisor: Zhu H B, pp. 36-60.) [7] 苏柳杭. 2018. CD163/TWEAK在动脉粥样硬化形成过程中作用机制的研究[D]. 硕士学位论文, 福建医科大学, 导师: 王焱, pp. 22-26. (Su L H.2018. Mechanism of CD163/TWEAK in the development of atherosclerosis[D]. Fujian Medical University, Supervisor: Wang Y, pp. 22-26.) [8] 吴常宝. 2021. CD163基因突变大白猪抗蓝耳病毒感染能力及其生产性能的研究[D]. 硕士学位论文, 内蒙古大学, 导师: 戴雁峰, pp. 33-71. (Wu C B.2021. Anti-PRRS virus infection capability and performance of the CD163 gene mutation large white pig[D]. Thesis for M.S., Inner Mongolia University, Supervisor: Dai Y F, pp. 33-71.) [9] 魏迎辉, 刘志国, 徐奎, 等. 2018. CD163双等位基因编辑猪的制备及传代[J]. 中国农业科学, 51: 770-777. (Wei Y H, Liu Z G, Xu K, et al.2018. Generation and propagation of cluster of differentiation 163 biallelic gene editing pigs[J]. Scientia Agricultura Sinica, 51: 770-777.) [10] 颜宇飞, 陈奇, 李灵筠, 等. 2019. 类风湿关节炎患者血清sCD163测定的临床价值[J]. 海军医学杂志, 40(1): 51-53. (Yan Y F, Chen Q, Li L Y, et al.2019. Clinical significance of serum sCD163 detection in patients with rheumatoid arthritis[J]. Journal of Navy Medicine, 40(1): 51-53.) [11] 于一涵. 2018. 巨噬细胞可溶性CD163与自身免疫疾病关系的研究进展[J]. 疑难病杂志, 17(2): 207-210. (Yu Y H.2018. Research progress of soluble CD163 in macrophage and it relationship with autoimmune diseases[J]. Chinese Journal of Difficult and Complicated Cases, 17(2): 207-210.) [12] 袁泓明. 2020. 基于基因编辑技术制备CD163基因修饰猪及α-Gal/Neu5Gc/Sda缺失猪[D]. 博士学位论文, 吉林大学, 导师: 欧阳红生, pp. 25-46. (Yuan H M.2020. Generation of CD163 modified pigs and α-Gal/Neu5Gc/Sda delete pigs with gene-editing technique[D]. Thesis for Ph.D., Jilin Universitity, Supervisor: Ou Y H S, pp. 25-46.) [13] 张莞悦, 赵斌, 钱行. 2023. 血清C5a、MMP-9、sCD163水平变化与系统性红斑狼疮患者SLEDAI评分的相关性及在其疾病转归预测中的应用[J]. 中国医学工程, 31(04): 126-129. (Zhang G Y, Zhao B, Qian X.2023. Correlation between changes in serum C5a, MMP-9, sCD163 levels and SLEDAI scores in patients with systemic lupus erythe[J]. China Medical Engineering, 31(04): 126-129.) [14] 张健, 吴珍芳, 杨化强. 2023. CD163基因敲除大白猪的抗蓝耳病性能和主要生产性能研究[J]. 华南农业大学学报, 44(03): 333-339. (Zhang J, Wu Z F, Yang H Q.2023. Resistance to blue ear disease and production performance assessment of CD163 gene-edited Large White pigs[J]. Journal of South China Agricultural University, 44(03): 333-339.) [15] 赵为民, 王慧利, 曹少先, 等. 2022. 猪CD163基因的单碱基编辑研究[J]. 畜牧兽医学报, 53(4): 1041-1050. (Zhao W M, Wang H L, Cao S X, et al.2022. The study of base editing of porcine CD163 gene[J]. Acta Veterinaria Et Zootechnica Sinica, 53(4): 1041-1050.) [16] Akahori H, Karmali V, Polavarapu R, et al.2015. CD163 interacts with TWEAK to regulate tissue regeneration after ischaemic injury[J]. Nature Communications, 6: 7792. [17] Bakogiannis C, Sachse M, Stamatelopoulos K, et al.2019. Platelet-derived chemokines in inflammation and atherosclerosis[J]. Cytokine(Philadelphia, Pa.), 122: 154157-154157. [18] Bover L C, Cardo-Vila M, Kuniyasu A, et al.2007. A previously unrecognized protein-protein interaction between TWEAK and CD163: Potential biological implications[J]. Journal of Immunology, 178(12): 8183-8194. [19] Burger B T, Beaton B P, Campbell M A, et al.2024. Generation of a commercial-scale founder population of Porcine reproductive and respiratory syndrome virus resistant pigs using CRISPR-Cas[J]. CRISPR Journal, 7(1): 12-28. [20] Burkard C, Lillico S G, Reid E, et al.2017. 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, 13(2): e1006206. [21] Burkard C, Opriessnig T, Mileham A J, et al.2018. Pigs lacking the scavenger receptor cysteine-rich domain 5 of CD163 are resistant to Porcine reproductive and respiratory syndrome virus 1 infection[J]. Journal of Virology, 92(16): e00415-18. [22] Calvert J G, Slade D E, Shields S L, et al.2007. CD163 expression confers susceptibility to Porcine reproductive and respiratory syndrome viruses[J]. Journal of Virology, 81(14): 7371-7379. [23] Chen J, Wang H, Bai J, et al.2019. Generation of pigs resistant to highly pathogenic-Porcine reproductive and respiratory syndrome virus through gene editing of CD163[J]. International Journal of Biological Sciences, 15(2): 481-492. [24] Das P B, Dinh P X, Ansari I H, et al.2010. The minor envelope glycoproteins GP2a and GP4 of Porcine reproductive and respiratory syndrome virus interact with the receptor CD163[J]. Journal of Virology, 84(4): 1731-1740. [25] Fabriek B O, Polfliet M M, Vloet R P, et al.2007. The macrophage CD163 surface glycoprotein is an erythroblast adhesion receptor[J]. Blood, 109(12): 5223-9. [26] Fabriek B O, van Bruggen R, Deng D M, et al.2009. The macrophage scavenger receptor CD163 functions as an innate immune sensor for bacteria[J]. Blood, 113(4): 887-92. [27] Finn A V, Nakano M, Polavarapu R, et al.2012. Hemoglobin directs macrophage differentiation and prevents foam cell formation in human atherosclerotic plaques[J]. Journal of the American College of Cardiology, 59(2): 166-177. [28] Fujiwara Y, Ohnishi K, Horlad H, et al.2020. CD163 deficiency facilitates lipopolysaccharide-induced inflammatory responses and endotoxin shock in mice[J]. Clinical & Translational Immunology, 9(9): e1162. [29] Graversen J H, Madsen M, Moestrup S K.2002. CD163: A signal receptor scavenging haptoglobin-hemoglobin complexes from plasma[J]. The International Journal of Biochemistry & Cell Biology, 34(4): 309-314. [30] Guo C, Wang M, Zhu Z, et al.2019. 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, 10: 1846. [31] Guo L, Niu J, Yu H, et al.2014. Modulation of CD163 expression by metalloprotease ADAM17 regulates Porcine reproductive and respiratory syndrome virus RNTRY[J]. Journal of Virology, 88(18): 10448-10458. [32] Gutierrez-Munoz C, Mendez-Barbero N, Svendsen P, et al.2020. CD163 deficiency increases foam cell formation and plaque progression in atherosclerotic mice[J]. The FASEB Journal, 34(11): 14960-14976. [33] Hogger P, Dreier J, Droste A, et al.1998. Identification of the integral membrane protein RM3/1 on human monocytes as a glucocorticoid-inducible member of the scavenger receptor cysteine-rich family(CD163)[J]. Journal of Immunology, 161(4): 1883-90. [34] Kowal K, Silver R, Sławińska E, et al.2011. CD163 and its role in inflammation[J]. Folia Histochemica et Cytobiologica, 49(3): 365-374. [35] Ma H, Jiang L, Qiao S, et al.2017. The crystal structure of the fifth scavenger receptor cysteine-rich domain of porcine CD163 reveals an important residue involved in Porcine reproductive and respiratory syndrome virus infection[J]. Journal of Virology, 91(3): e01897-16. [36] Ma H, Li R, Jiang L, et al.2021. Structural comparison of CD163 SRCR5 from different species sheds some light on its involvement in porcine reproductive and respiratory syndrome virus-infection in vitro[J]. Veterinary Research, 52(1): 97. [37] Madsen M, Moller H J, Nielsen M J, et al.2004. Molecular characterization of the haptoglobin.hemoglobin receptor CD163. Ligand binding properties of the scavenger receptor cysteine-rich domain region[J]. Journal of Biological Chemistry, 279(49): 51561-51567. [38] Moestrup S K, Kristiansen M, Graversen J H, et al.2001. Identification of the haemoglobin scavenger receptor[J]. Nature(London), 409(6817): 198-201. [39] Moniuszko M, Kowal K, Rusak M, et al.2006. Monocyte CD163 and CD36 expression in human whole blood and isolated mononuclear cell samples: Influence of different anticoagulants[J]. Clinical and Vaccine Immunology, 13(6): 704-707. [40] Nesbitt C, Galina Pantoja L, Beaton B, et al.2024. Pigs lacking the SRCR5 domain of CD163 protein demonstrate heritable resistance to the PRRS virus and no changes in animal performance from birth to maturity[J]. Frontiers in Genome Editing, 6: 1322012. [41] Nielsen M J, Madsen M, Moller H J, et al.2006. The macrophage scavenger receptor CD163: Endocytic properties of cytoplasmic tail variants[J]. Journal of Leukocyte Biology, 79(4): 837-845. [42] Radzun H J, Kreipe H, Bodewadt S, et al.1987. Ki-M8 monoclonal antibody reactive with an intracytoplasmic antigen of monocyte/macrophage lineage[J]. Blood, 69(5): 1320-1327. [43] Ratajczak W, Atkinson S D, Kelly C.2022. The TWEAK/Fn14/CD163 axis-implications for metabolic disease[J]. Reviews in Endocrine & Metabolic Disorders, 23(3): 449-462. [44] Sachais B S, Turrentine T, Dawicki M J, et al.2007. Elimination of platelet factor 4(PF4) from platelets reduces atherosclerosis in C57Bl/6 and apoE-/- mice[J]. Thrombosis and Haemostasis, 98(5): 1108-1113. [45] Salgado B, Rivas R B, Pinto D, et al.2024. Genetically modified pigs lacking CD163 PSTII-domain-coding exon 13 are completely resistant to PRRSV infection[J]. Antiviral Research, 221: 105793. [46] Sarabi A, Kramp B K, Drechsler M, et al.2011. CXCL4L1 inhibits angiogenesis and induces undirected endothelial cell migration without affecting endothelial cell proliferation and monocyte recruitment[J]. Journal of Thrombosis and Haemostasis, 9(1): 209-219. [47] Schaer C A, Schoedon G, Imhof A, et al.2006. Constitutive endocytosis of CD163 mediates hemoglobin-heme uptake and determines the noninflammatory and protective transcriptional response of macrophages to hemoglobin[J]. Circulation Research, 99(9): 943-500. [48] Siwan E, Twigg S M, Min D.2022. Alterations of CD163 expression in the complications of diabetes: A systematic review[J]. Journal of Diabetes and its Complications, 36(4): 108150. [49] Skytthe M K, Graversen J H, Moestrup S K.2020. Targeting of CD163(+) macrophages in inflammatory and malignant diseases[J]. International Journal of Molecular Sciences, 21(15): 5497. [50] Sulahian T H, Högger P, Wahner A E, et al.2000. Human monocytes express CD163, which is upregulated by il-10 and identical to p155[J]. Cytokine, 12(9): 1312-1321. [51] Van den Heuvel M M, Tensen C P, van As J H, et al.1999. Regulation of CD163 on human macrophages: Cross-linking of CD163 induces signaling and activation[J]. Journal of Leukocyte Biology, 66(5): 858-866. [52] Van Gorp H, Delputte P L, Nauwynck H J.2010a. Scavenger receptor CD163, a Jack-of-all-trades and potential target for cell-directed therapy[J]. Molecular Immunology, 47(7-8): 1650-1660. [53] Van Gorp H, Van Breedam W, Delputte P L, et al.2010b. Identification of the CD163 protein domains involved in infection of the Porcine reproductive and respiratory syndrome virus[J]. Journal of Virology, 84(6): 3101-3105. [54] Van Gorp H, Van Breedam W, Delputte P L, et al.2008. Sialoadhesin and CD163 join forces during entry of the Porcine reproductive and respiratory syndrome virus[J]. Journal of General Virology, 89(12): 2943-2953. [55] Varga G, Ehrchen J, Tsianakas A, et al.2008. Glucocorticoids induce an activated, anti-inflammatory monocyte subset in mice that resembles myeloid-derived suppressor cells[J]. Journal of Leukocyte Biology, 84(3): 644-650. [56] Wells K D, Bardot R, Whitworth K M, et al.2017. Replacement of porcine CD163 scavenger receptor cysteine-rich domain 5 with a CD163-like homolog confers resistance of pigs to genotype 1 but not genotype 2 Porcine reproductive and respiratory syndrome virus[J]. Journal of Virology, 91(2): e01521-16. [57] Wang H, Shen L, Chen J, et al.2019. Deletion of CD163 exon 7 confers resistance to highly pathogenic porcine reproductive and respiratory viruses on pigs[J]. International Journal of Biological Sciences, 15(9): 1993-2005. [58] Whitworth K M, Lee K, Benne J A, et al.2014. Use of the CRISPR/Cas9 system to produce genetically engineered pigs from in vitro-derived oocytes and embryos[J]. Biology of Reproduction, 91(3): 78. [59] Whitworth K M, Rowland R R, Ewen C L, et al.2016. Gene-edited pigs are protected from Porcine reproductive and respiratory syndrome virus[J]. Nature Biotechnology, 34(1): 20-22. [60] Williams L, Jarai G, Smith A, et al.2002. IL-10 expression profiling in human monocytes[J]. Journal of leukocyte biology, 72(4): 800-809. [61] Xu K, Zhou Y, Mu Y, et al.2020. 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, 9: e57132. [62] Xu K, Zhou Y, Shang H, et al.2023. Pig macrophages with site-specific edited CD163 decrease the susceptibility to infection with Porcine reproductive and respiratory syndrome virus[J]. Journal of Integrative Agriculture, 22(7): 2188-2199. [63] Yang H, Zhang J, Zhang X, et al.2018. CD163 knockout pigs are fully resistant to highly pathogenic Porcine reproductive and respiratory syndrome virus[J]. Antiviral Research, 151: 63-70.
