Heterogeneity of porcine bone marrow-derived dendritic cells induced by GM-CSF

Abstract

In vitro generation of dendritic cells (DCs) is advantageous for overcoming the low frequency of primary DCs and the difficulty of applying isolation techniques for studying DC immunobiology. The culture of bone marrow cells with granulocyte-macrophage colony-stimulating factor (GM-CSF) has been used extensively to generate bone marrow-derived dendritic cells (BMDCs). Studies have reported the heterogeneity of cells grown in murine GM-CSF culture based on the levels of MHCII expression. Although porcine DCs are generated by this classical method, the exact characteristics of the BMDC population have not yet been defined. In this study, we discriminated GM-CSF-grown BMDCs from gnotobiotic miniature pigs according to several criteria including morphology, phenotype, gene expression pattern and function. We showed that porcine BMDCs were heterogeneous cells that differentially expressed MHCII. MHCIIhigh cells displayed more representative of DC-like morphology and phenotype, including costimulatory molecules, as well as they showed a superior T cell priming capacity as compared to MHCIIlow cell. Our data showed that the difference in MHCIIhigh and MHCIIlow cell populations involved distinct maturation states rather than the presence of different cell types. Overall, characterization of porcine BMDC cultures provides important information about this widely used cellular model.

Immunomodulatory effect of probiotic exopolysaccharides in a porcine in vitro co-culture model mimicking the intestinal environment on ETEC infection

“…On day 5 of culture an immatureimmunophenotype of the moDCs was obtained however after 48 h of cultivation with LPS from E. coli O111:B4, we observed an increase in surface marker expression for the MHC class II, CD80/86 surface marker (> 45%). Similar results were obtained other available studies which investigated the ability of live pathogens or their PAMPs to induce the maturation and activation of monocyte-derived macrophages and dendritic cells, with similar results (Carrasco et al 2001 ; Kim et al 2019 ; Meital 2019 ).…”

Heterogeneous populations from in vitro cultures of antigen presenting cells in pigs

“…On the contrary, our cultures were treated with GM-CSF and IL-4, producing MHC-II mid cells that resemble DCs and MHC-II high cells that resemble Macs, according to their immunophenotype, phagocytosis capacity and the expression of signature genes from the DC or Macs lineage. A previous work described that different maturation states were obtained when swine BM precursors were cultured only with GM-CSF (Kim et al, 2019). In pigs, same as in other species, DC maturation is associated with an increase in MHC-II expression, in addition to loss of CD14 expression and decreased phagocytic capacity (Singleton et al, 2016), reason why different states of maturation were are not consistent with the cell cultures analyzed in this work.…”

Tolerogenic dendritic cells alleviate collagen‐induced arthritis by forming microchimerism and affecting the expression of immune checkpoint molecules

“…The morphology of the cells on the sixth day of culture was observed under a microscope, and obvious cell colonies were observed in each group of cells. The formation of colonies was a sign of the successful induction of monocyte‐derived DCs by GM‐CSF [28]. In addition, the positive rate of rat DC‐specific marker OX‐62 detected by flow cytometry was >95%, which also indicated that the induced DCs had high purity and met the requirements of subsequent experiments.…”