IP Library › Granted Patent US 12,473,341
Granted Patent B2
US 12,473,341 · App. 18/758,151 · Granted Nov 18, 2025

Methods and compositions for treating and protecting against porcine reproductive and respiratory syndrome virus

Inventors: Raymond R.R. Rowland (Chamgpaign, IL); Ana Stoian (Manhattan, KS)
Assignee: Kansas State University Research Foundation
C07K14/705A01K67/027C12N15/85
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,473,341
App. No.
18/758,151
Granted
Nov 18, 2025
Kind
B2
Abstract

The present disclosure provides compositions and methods for decreasing permissiveness to infection by PRRSV. The methods involve inserting peptides or oligopeptides into the CD163 SRCR domain 5 to disrupt the protein structure and thereby prevent PRRSV infection. The deletion of the exon 13 region of PSTII eliminates infection. The replacement of cysteine residues with alanines in SRCR5 as a means to disrupt disulfide bond formation. The final method is the deletion of, or amino acid substitution within the SRCR4-5 interdomain region.

Claims (10)

1 . A cell comprising a modification in exon 13 of a CD163 gene, wherein the cell does not comprise a disruption of any other exon of the CD163 gene.

2 . The cell of claim 1 , wherein the modification comprises a complete deletion of exon 13.

3 . The cell of claim 1 , wherein the modification is in a genome of the cell.

4 . The cell of claim 1 , wherein the modification results in a reduction in susceptibility to infection by PRRSV relative to that of a cell that does not comprise the modification.

5 . The cell of claim 4 , wherein the reduction in susceptibility is at least 50% in comparison to that of the cell that does not comprise the modification.

6 . A cell comprising a modification in exon 13 of a CD163 gene, wherein the cell does not comprise a disruption of exon 14 of the CD163 gene.

7 . The cell of claim 6 , wherein the modification comprises a complete deletion of exon 13.

8 . The cell of claim 6 , wherein the modification is in a genome of the cell.

9 . The cell of claim 6 , wherein the modification results in a reduction in susceptibility to infection by PRRSV relative to that of a cell that does not comprise the modification.

10 . The cell of claim 9 , wherein the reduction in susceptibility is at least 50% relative to that of the cell that does not comprise the modification.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 13, 2025
From: ROWLAND, RAYMOND R.R.; STOIAN, ANA
To: KANSAS STATE UNIVERSITY RESEARCH FOUNDATION
Reel/Frame 069837/0026 →
Continuity (3)
Continuation 16756756
Provisional Application 62573078 · Oct 16, 2017
Related Publication 20240343774A1 · Oct 17, 2024
References Cited (76)
US 1008035A · Hester · 1911 [cited by applicant]
US 9820475B2 · Prather · 2017 [cited by applicant]
US 9854790B2 · Ait-Ali · 2018 [cited by applicant]
US 10091975B2 · Prather · 2018 [cited by applicant]
US 10405526B2 · Prather · 2019 [cited by applicant]
US 10827730B2 · Prather · 2020 [cited by applicant]
US 11019809B2 · Prather · 2021 [cited by applicant]
US 11160260B2 · Prather · 2021 [cited by applicant]
US 12060403B2 · Rowland · 2024 [cited by examiner]
US 20110016546A1 · Bedell · 2011 [cited by applicant]
CN 101503688A · 2009 [cited by applicant]
WO 2019079285A1 · 2019 [cited by applicant]
WO 2020198541A1 · 2020 [cited by applicant]
Van Gorp, H., Van Breedam, W., Van Doorsselaere, J., Delputte, P.L. and Nauwynck, H.J., 2010. Identification of the CD163 protein domains involved in infection of the porcine reproductive and respiratory syndrome virus.… [cited by examiner]
Burkard, Christine, 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 fun… [cited by examiner]
Wang, et al. Immune Responses in Piglets Infected with Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus. Veterinary Immunology and Immunopathology. vol. 142, Issue 3-4 (2011): 9 pages. [cited by applicant]
Wang, et al. PK-15 Cells Transfected with Porcine CD163 by PiggyBac Transposon System are Susceptible to Porcine Reproductive and Respiratory Syndrome Virus. Journal of Virological Methods. vol. 193, Issue 2 (2013): 8 p… [cited by applicant]
Wells, et al. 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 Vi… [cited by applicant]
Wensvoort, et al. Mystery Swine Disease in The Netherlands: The Isolation of Lelystad Virus. Veterinary Quarterly. vol. 13, Issue 3 (1991): 10 pages. [cited by applicant]
Whitworth, et al. Gene-edited Pigs are Protected from Porcine Reproductive and Respiratory Syndrome Virus. Nature Biotechnology. vol. 34, Issue 1 (2016): 3 pages. [cited by applicant]
Whitworth, et al. Use of the CRISPR/Cas9 System to Produce Genetically Engineered Pigs from in Vitro-derived Oocytes and Embryos. Biology of Reproduction. vol. 91, Issue 3 (2014): 13 pages. [cited by applicant]
Workman, et al. Evaluating Large Spontaneous Deletions in a Bovine Cell Line Selected for Bovine Viral Diarrhea Virus Resistance. Viruses. vol. 13 (2021): 17 pages. [cited by applicant]
Yang, et al. CD163 Knockout Pigs are Fully Resistant to Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus. Antiviral Research. vol. 151 (2018): 63-70. [cited by applicant]
Yu, et al. CD163ASRCR5 MARC-145 Cells Resist PRRSV-2 Infection via Inhibiting Virus Uncoating, Which Requires the Interaction of CD163 With Calpain 1. Frontiers in Microbiology. vol. 10 (2020): 16 pages. [cited by applicant]
Zhu, et al. TREM2 Suppresses the Proinflammatory Response to Facilitate PRRSV Infection Via PI3K/NF-κB Signaling. PLoS Pathogens. vol. 16, Issue 5 (2020): 28 pages. [cited by applicant]
Zimmerman, et al. Porcine Reproductive and Respiratory Syndrome Viruses (Porcine Arteriviruses). Diseases of Swine (2019): 24 pages. [cited by applicant]
Burkard et al. Pigs Lacking the Scavenger Receptor Cysteine-Rich Domain 5 of CD163 Are Resistant to Porcine Reproductive and Respiratory Syndrome Virus 1 Infection. Journal of Virology, vol. 92, Issue 16, e00415-18 (Aug… [cited by applicant]
Calvert et al. CD163 Expression Confers Susceptibility to Porcine Reproductive and Respiratory Syndrome Viruses. Journal of Virology, vol. 81, No. 14, pp. 7371-7379 (Jul. 2007). Published ahead of print May 9, 2007. [cited by applicant]
Dong et al. Associations of natural variation in the CD163 and other candidate genes on host response of nursery pigs to porcine reproductive and respiratory syndrome virus infection. Journal of Animal Science, 2021, vo… [cited by applicant]
GenBank Accession No. AJ311716. Version No. AJ311716.1. Sus scrofa mRNA for putative CD163 antigen. Record created Jan. 7, 2003. 2 pages. Retrieved Nov. 20, 2023 at URL: https://www.ncbi.nlm.nih.gov/nuccore/AJ311716.1/. [cited by applicant]
GenBank Accession No. EU016226. Version No. EU016226.1. Sus scrofa CD163 mRNA, complete cds. Record created Sep. 4, 2008. 2 pages. Retrieved Nov. 20, 2023 at URL: https://www.ncbi.nlm.nih.gov/nuccore/EU016226.1/. [cited by applicant]
Graversen et al. CD163: a signal receptor scavenging haptoglobin-hemoglobin complexes from plasma. The International Journal of Biochemistry & Cell Biology 34 (2002) 309-314. [cited by applicant]
Guo et al. Modulation of CD163 Expression by Metalloprotease ADAM17 Regulates Porcine Reproductive and Respiratory Syndrome Virus Entry. Journal of Virology, vol. 88, No. 18, pp. 10448-10458 (Sep. 2014). Published ahead… [cited by applicant]
Ma et al. 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. Journal of Vi… [cited by applicant]
PCT/US2018/056065 International Search Report and Written Opinion dated Feb. 25, 2019. [cited by applicant]
Van Gorp et al. Sialoadhesin and CD163 join forces during entry of the porcine reproductive and respiratory syndrome virus. Journal of General Virology (2008), 89, 2943-2953. [cited by applicant]
Allende, et al. North American and European Porcine Reproductive and Respiratory Syndrome Viruses Differ in Non-structural Protein Coding Regions. Journal of General Virology. vol. 80 (1999): 9 pages. [cited by applicant]
Benfield, et al. Characterization of Swine Infertility and Respiratory Syndrome (SIRS) Virus (isolate ATCC VR-2332). Journal of Veterinary Diagnostic Investigation. vol. 4, Issue 2 (1992): 7 pages. [cited by applicant]
Bullido, et al. Monoclonal Antibodies Specific for Porcine Monocytes/Macrophages: Macrophage Heterogeneity in the Pig Evidenced by the Expression of Surface Antigens. Tissue Antigens. vol. 49, Issue 4 (1997): 11 pages. [cited by applicant]
Burkard, 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. PLoS … [cited by applicant]
Cafruny, et al. Antibiotic-Mediated Inhibition of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Infection: A Novel Quinolone Function Which Potentiates the Antiviral Cytokine Response in MARC-145 Cells and… [cited by applicant]
Chen et al. Generation of Pigs Resistant to Highly Pathogenic-Porcine Reproductive and Respiratory Syndrome Virus Through Gene Editing of CD163. International Journal of Biological Sciences. vol. 15, Issue 2 (2019): 12 … [cited by applicant]
Collins, et al. Isolation of Swine Infertility and Respiratory Syndrome Virus (isolate ATCC VR-2332) in North America and Experimental Reproduction of the Disease in Gnotobiotic Pigs. Journal of Veterinary Diagnostic In… [cited by applicant]
Duan, et al. Virus Quantification and Identification of Cellular Targets in the Lungs and Lymphoid Tissues of Pigs at Different Time Intervals after Inoculation with Porcine Reproductive and Respiratory Syndrome Virus (… [cited by applicant]
Etzerodt, et al. Structural Basis for Inflammation-driven Shedding of CD163 Ectodomain and Tumor Necrosis Factor-α in Macrophages. Journal of Biological Chemistry. vol. 289, Issue 2 (2014): 11 pages. [cited by applicant]
Fabriek, et al. The Macrophage CD163 Surface Glycoprotein is an Erythroblast Adhesion Receptor. Blood. vol. 109, Issue 12 (2007): 7 pages. [cited by applicant]
Fabriek, et al. The Macrophage Scavenger Receptor CD163. Immunobiology. vol. 210, Issue 2-4 (2005): 8 pages. [cited by applicant]
Guo, 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. Frontiers in Immunolo… [cited by applicant]
Halbur, et al. Comparative Pathogenicity of Nine US Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Isolates in a Five-week-old Cesarean-derived, Colostrum-deprived Pig Model. Journal of Veterinary Diagnosti… [cited by applicant]
Halbur, et al. Update on Abortion Storms and Sow Mortality. Swine Health and Production. vol. 5, Issue 2 (1997): 1 page. [cited by applicant]
Kappes, et al. PRRSV Structure, Replication and Recombination: Origin of Phenotype and Genotype Diversity. Virology (2015). 12 pages. [cited by applicant]
Kittawornrat, et al. Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Surveillance using Pre-weaning Oral Fluid Samples Detects Circulation of Wild-type PRRSV. Veterinary Microbiology. vol. 168, Issue 2-4 (20… [cited by applicant]
Kristiansen, et al. Identification of the Haemoglobin Scavenger Receptor. Nature. vol. 409, Issue 6817 (2001): 4 pages. [cited by applicant]
Kuhn, et al. Reorganization and Expansion of the Nidoviral Family Arteriviridae. Archives of Virology. vol. 161, Issue 3 (2016): 14 pages. [cited by applicant]
Ladinig, et al. Pathogenicity of Three Type 2 Porcine Reproductive and Respiratory Syndrome Virus Strains in Experimentally Inoculated Pregnant Gilts. Virus Research. vol. 203 (2015): 12 pages. [cited by applicant]
Lamontagne, et al. Porcine Reproductive and Respiratory Syndrome Virus Persistence in Blood, Spleen, Lymph Nodes, and Tonsils of Experimentally Infected Pigs Depends on the Level of CD8high T Cells. Viral Immunology. vo… [cited by applicant]
Law, et al. A New Macrophage Differentiation Antigen Which is a Member of the Scavenger Receptor Superfamily. European Journal of Immunology. vol. 23, Issue 9 (1993): 6 pages. [cited by applicant]
Lee, et al. A DNA-launched Reverse Genetics System for Porcine Reproductive and Respiratory Syndrome Virus Reveals that Homodimerization of the Nucleocapsid protein is Essential for Virus Infectivity. Virology. vol. 331… [cited by applicant]
Lee, et al. Generation of a Porcine Alveolar Macrophage Cell Line for the Growth of Porcine Reproductive and Respiratory Syndrome Virus. Journal of Virological Methods. vol. 163, Issue 2 (2010): 6 pages. [cited by applicant]
Madson, et al. Molecular Characterization of the Haptoglobin.hemoglobin Receptor CD163. Ligand Binding Properties of the Scavenger Receptor Cysteine-rich Domain Region. vol. 279, Issue 49 (2004): 8 pages. [cited by applicant]
Meulenberg, et al. Lelystad Virus, The Causative Agent of Porcine Epidemic Abortion and Respiratory Syndrome (PEARS), Is related to LDV and EAV. Virology. vol. 192, Issue 1 (1993): 11 pages. [cited by applicant]
Nelson, et al. Porcine Reproductive and Respiratory Syndrome Virus Comparison: Divergent Evolution on Two Continents. Journal of virology. vol. 73, Issue 1 (1999): 11 pages. [cited by applicant]
Onofre, et al. Scavenger Receptor CD163 and its Biological Functions. Acta Medica (Hradec Kralove). vol. 52, Issue 2 (2009): 5 pages. [cited by applicant]
Pasternak, et al. Development and Application of a Porcine Specific ELISA for the Quantification of Soluble CD163. Veterinary Immunology and Immunopathology. vol. 210 (2019): 8 pages. [cited by applicant]
Patton, et al. Modulation of CD163 Receptor Expression and Replication of Porcine Reproductive and Respiratory Syndrome Virus in Porcine Macrophages. Virus research. vol. 140, Issue 1-2 (2009): 11 pages. [cited by applicant]
Pei, et al. Porcine Reproductive and Respiratory Syndrome Virus as a Vector: Immunogenicity of Green Fluorescent Protein and Porcine Circovirus Type 2 Capsid Expressed from Dedicated Subgenomic RNAs. Virology. vol. 389.… [cited by applicant]
Popescu, et al. Genetically Edited Pigs Lacking CD163 Show No Resistance Following Infection with the African Swine Fever Virus Isolate, Georgia Jan 2007. Virology. vol. 501 (2017): 5 pages. [cited by applicant]
Prather, et al. Knockout of Maternal CD163 Protects Fetuses from Infection with Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). Scientific Reports. vol. 7, Issue 1 (2017): 5 pages. [cited by applicant]
Ritter, et al. Genomic Organization and Chromosomal Localization of the Human CD163 (M130) Gene: A Member of the Scavenger Receptor Cysteine-rich Superfamily. Biochemical and Biophysical Research Communications. vol. 26… [cited by applicant]
Sanchez-Torres, et al. Expression of Porcine CD163 on Monocytes/Macrophages Correlates with Permissiveness to African Swine Fever Infection. Archives of Virology. vol. 148, Issue 12 (2003): 17 pages. [cited by applicant]
Snijder, et al. The Molecular Biology of Arteriviruses. Journal of General Virology. vol. 79 (1998): 19 pages. [cited by applicant]
Stoian, et al. Identification of CD163 Regions that are Required for Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Infection but not for Binding to Viral Envelope Glycoproteins. Virology. vol. 574 (2022): … [cited by applicant]
Stoian, et al. Mutations within Scavenger Receptor Cysteine-rich (SRCR) Protein Domain 5 of Porcine CD163 Involved in Infection with Porcine Reproductive and Respiratory Syndrome Virus (PRRS). Journal of General Virolog… [cited by applicant]
Subramanian, et al. CD163 and IgG Codefend Against Cytotoxic Hemoglobin via Autocrine and Paracrine Mechanisms. The Journal of Immunology. vol. 190, Issue 10 (2013): 12 pages. [cited by applicant]
Van Gorp, et al. Scavenger Receptor CD163, A Jack-of-all-trades and Potential Target for Cell-directed Therapy. Molecular Immunology. vol. 47, Issue 7-8 (2010): 11 pages. [cited by applicant]
Wang, et al. Emergence of a Virulent Porcine Reproductive and Respiratory Syndrome Virus in Vaccinated Herds in the United States. Virus Research. vol. 210 (2015): 34-41. [cited by applicant]