Journal Article

Novel Engraftment and T Cell Differentiation of Human Hematopoietic Cells in ART−/−IL2RG−/Y SCID Pigs

Snella, Elizabeth M., Adur, Malavika, Kiefer, Zoe E., Boettcher, Adeline N., Westin, Erik R., Jens, Jackie, Putz, Ellie J., Tuggle, Christopher K., Charley, Sara E., Wiarda, Jayne E., Swanson, Jack J., Dell'Anna, Giuseppe, Cino-Ozuna, A. Giselle, Ho, Chak-Sum (Sam), Dekkers, Jack C. M., Byrne, Kristen A., Schultz, Blythe, Cunnick, Joan E., Loving, Crystal L., Goldman, Frederick, Sathe, Swanand, Ahrens, Amanda P., Li, Yunsheng, Kiupel, Matti, Ross, Jason W.

2020

Overview

Pigs with severe combined immunodeficiency (SCID) are an emerging biomedical animal model. Swine are anatomically and physiologically more similar to humans than mice, making them an invaluable tool for preclinical regenerative medicine and cancer research. One essential step in further developing this model is the immunological humanization of SCID pigs. In this work we have generated T B NK SCID pigs through site directed CRISPR/Cas9 mutagenesis of within a naturally occurring genetic background. We confirmed pigs lacked T, B, and NK cells in both peripheral blood and lymphoid tissues. Additionally, we successfully performed a bone marrow transplant on one male SCID pig with bone marrow from a complete swine leukocyte antigen (SLA) matched donor without conditioning to reconstitute porcine T and NK cells. Next, we performed injections of cultured human CD34 selected cord blood cells into the fetal SCID pigs. At birth, human CD45 CD3ε cells were detected in cord and peripheral blood of injected SCID piglets. Human leukocytes were also detected within the bone marrow, spleen, liver, thymus, and mesenteric lymph nodes of these animals. Taken together, we describe critical steps forwards the development of an immunologically humanized SCID pig model.

• Publisher: Frontiers Media SA,Frontiers Media S.A
• Subject: Animal models / Animals / Animals, Genetically Modified / Antigens, CD34 / biomedical model / Blood cells / Bone marrow / Bone Marrow Transplantation / CD34 antigen / CD45 antigen / Cell Differentiation / Cesarean section / Chimera / Cord blood / CRISPR / CRISPR-Cas Systems / Disease Models, Animal / DNA-Binding Proteins - deficiency / Embryos / Fetuses / Fibroblasts / Gene Targeting / Genes / Genetic Engineering / Genetic testing / Genotype & phenotype / Graft Survival / Hematopoietic stem cells / Hogs / Host vs Graft Reaction / humanization / Humans / Immunology / Interleukin Receptor Common gamma Subunit - genetics / Killer Cells, Natural / Laboratory animals / Leukocytes / Lymph nodes / Lymphocytes T / Lymphoid tissue / Models, Animal / Mutagenesis / Mutation / Peripheral blood / Polymorphism / Regenerative medicine / SCID / Severe combined immunodeficiency / Severe Combined Immunodeficiency - genetics / Stem cells / Swine / T-Lymphocytes - metabolism / Transplantation, Heterologous