Development of Pig Conventional Dendritic Cells From Bone Marrow Hematopoietic Cells in vitro

• Published in: Frontiers in immunology 2020-10, Vol.11, p.553859-553859
• Main Authors: Li, Yanli, Puebla-Clark, Lucinda, Hernández, Jesús, Díaz, Ivan, Mateu, Enric
• Format: Article
• Language: English
• Subjects: Animals; Antigens, Differentiation - immunology; Bone Marrow - immunology; CD14+ DC; cDC1; cDC2; Cell Differentiation - immunology; Cytokines - immunology; Dendritic Cells - cytology; Dendritic Cells - immunology; Flt3L; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - immunology; Immunology; pig; Swine
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2020.553859

In recent years, porcine dendritic cells (DCs) have been identified from pig tissues. However, studying the interaction of porcine DCs with pathogens is still difficult due to the scarcity of DCs in tissues. In the present work, the Flt3-ligand (Flt3L)-based derivation system was further characterized and compared with other cytokine derivation models using a combination of factors: stem cell factor (SCF), GM-CSF, and IL-4. The method using Flt3L alone or combined with SCF supported the development of pig bone marrow hematopoietic cells into equivalent conventional DCs (cDCs). The equivalent cDC1 (the minor population in the cultures) were characterized as CADM1 CD14 MHC-II CD172a CD1 CD163 DEC205 CD11R3 CD11R1 CD33 CD80/86 . They expressed high levels of FLT3, ZBTB46, XCR1, and IRF8 mRNA, were efficient in endocytosing dextran and in proliferating allogenic CD4 CD8 T cells, but were deficient in phagocyting inactivated ( ). Also, after poly I:C stimulation, they predominantly produced IL-12p40a and matured as indicated by the increase of MHC-I, MHC-II, and CD80/86. The equivalent cDC2 (the main population) were CADM1 CD14 MHC-II C D172a CD1 CD163 DEC205 CD11R3 CD11R1 CD33 CD80/86 ; meanwhile, they overexpressed FcεR1α and IRF4 mRNA. They showed high efficiency in the endocytosis of dextran, but weak in phagocytosing bacteria. They supported allogenic CD4 CD8 /CD4 CD8 T cell proliferation and were high producers of IL-12p40 (upon TLR7 stimulation) and IL-10 (upon TLR7 stimulation). TLR ligand stimulation also induced their maturation. In addition, a CD14 population was identified with the phenotype CADM1 CD14 MHC-II CD172a CD1 CD163 DEC205 CD11R3 CD11R1 CD33 CD80/86 . They shared some functional similarities with cDC2 and were distinguishable from macrophages. This CD14 population was efficient in phagocyting but showed less maturation upon TLR ligand stimulation than cDC1 or cDC2. The alternative methods of DC derivation including GM-CSF and/or IL-4 produced mostly CADM1 cells that did not fulfill the canonical phenotype of porcine DCs. Our study provides an exhaustive characterization of Flt3L-derived DCs with different methods that can help the study of the interaction of DCs with porcine-relevant pathogens.