Porcine signal regulatory protein alpha binds to human CD47 to inhibit phagocytosis: Implications for human hematopoietic stem cell transplantation into severe combined immunodeficient pigs
Adeline N. Boettcher
Department of Animal Science, Iowa State University, Ames, Iowa
Search for more papers by this authorJoan E. Cunnick
Department of Animal Science, Iowa State University, Ames, Iowa
Search for more papers by this authorEllis J. Powell
Department of Animal Science, Iowa State University, Ames, Iowa
National Animal Disease Center, Ruminant Diseases and Immunology Unit, US Department of Agriculture, Agricultural Research Service, Ames, Iowa
Search for more papers by this authorTimothy K. Egner
Department of Chemistry, Iowa State University, Ames, Iowa
Search for more papers by this authorSara E. Charley
Department of Animal Science, Iowa State University, Ames, Iowa
Search for more papers by this authorCrystal L. Loving
National Animal Disease Center, Food Safety and Enteric Pathogens Unit, US Department of Agriculture, Agricultural Research Service, Ames, Iowa
Search for more papers by this authorCorresponding Author
Christopher K. Tuggle
Department of Animal Science, Iowa State University, Ames, Iowa
Correspondence
Christopher K. Tuggle, Department of Animal Science, Ames, IA.
Email: [email protected]
Search for more papers by this authorAdeline N. Boettcher
Department of Animal Science, Iowa State University, Ames, Iowa
Search for more papers by this authorJoan E. Cunnick
Department of Animal Science, Iowa State University, Ames, Iowa
Search for more papers by this authorEllis J. Powell
Department of Animal Science, Iowa State University, Ames, Iowa
National Animal Disease Center, Ruminant Diseases and Immunology Unit, US Department of Agriculture, Agricultural Research Service, Ames, Iowa
Search for more papers by this authorTimothy K. Egner
Department of Chemistry, Iowa State University, Ames, Iowa
Search for more papers by this authorSara E. Charley
Department of Animal Science, Iowa State University, Ames, Iowa
Search for more papers by this authorCrystal L. Loving
National Animal Disease Center, Food Safety and Enteric Pathogens Unit, US Department of Agriculture, Agricultural Research Service, Ames, Iowa
Search for more papers by this authorCorresponding Author
Christopher K. Tuggle
Department of Animal Science, Iowa State University, Ames, Iowa
Correspondence
Christopher K. Tuggle, Department of Animal Science, Ames, IA.
Email: [email protected]
Search for more papers by this authorAbstract
Background
Severe combined immunodeficient (SCID) pigs are an emerging animal model being developed for biomedical and regenerative medicine research. SCID pigs can successfully engraft human-induced pluripotent stem cells and cancer cell lines. The development of a humanized SCID pig through xenotransplantation of human hematopoietic stem cells (HSCs) would be a further demonstration of the value of such a large animal SCID model. Xenotransplantation success with HSCs into non-obese diabetic (NOD)-derived SCID mice is dependent on the ability of NOD mouse signal regulatory protein alpha (SIRPA) to bind human CD47, inducing higher phagocytic tolerance than other mouse strains. Therefore, we investigated whether porcine SIRPA binds human CD47 in the context of developing a humanized SCID pig.
Methods
Peripheral blood mononuclear cells (PBMCs) were collected from SCID and non-SCID pigs. Flow cytometry was used to assess whether porcine monocytes could bind to human CD47. Porcine monocytes were isolated from PBMCs and were subjected to phagocytosis assays with pig, human, and mouse red blood cell (RBC) targets. Blocking phagocytosis assays were performed by incubating human RBCs with anti-human CD47 blocking antibody B6H12, non-blocking antibody 2D3, and nonspecific IgG1 antibody and exposing to human or porcine monocytes.
Results
We found that porcine SIRPA binds to human CD47 in vitro by flow cytometric assays. Additionally, phagocytosis assays were performed, and we found that porcine monocytes phagocytose human and porcine RBCs at significantly lower levels than mouse RBCs. When human RBCs were preincubated with CD47 antibodies B6H12 or 2D3, phagocytosis was induced only after B6H12 incubation, indicating the lower phagocytic activity of porcine monocytes with human cells requires interaction between porcine SIRPA and human CD47.
Conclusions
We have shown the first evidence that porcine monocytes can bind to human CD47 and are phagocytically tolerant to human cells, suggesting that porcine SCID models have the potential to support engraftment of human HSCs.
Supporting Information
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| xen12466-sup-0001-FigS1.tifTIFF image, 277.9 KB | |
| xen12466-sup-0002-FigS2.tifTIFF image, 132.5 KB | |
| xen12466-sup-0003-Legends.docxWord document, 15.2 KB |
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