Comparative analysis of canine monocyte- and bone-marrow-derived dendritic cells

Abstract

Dendritic cells (DC) represent a heterogeneous cell family of major importance for innate immune responses against pathogens and antigen presentation during infection, cancer, allergy and autoimmunity. The aim of the present study was to characterize canine DC generated in vitro with respect to their phenotype, responsiveness to toll-like receptor (TLR) ligands and T-cell stimulatory capacity. DC were derived from monocytes (MoDC) and from bone marrow hematopoietic cells cultured with either Flt3-ligand (FL-BMDC) or with GM-CSF (GM-BMDC). All three methods generated cells with typical DC morphology that expressed CD1c, CD11c and CD14, similar to macrophages. However, CD40 was only found on DC, CD206 on MΦ and BMDC, but not on monocytes and MoDC. CD1c was not found on monocytes but on all in vitro differentiated cells. FL-BMDC and GM-BMDC were partially positive for CD4 and CD8. CD45RA was expressed on a subset of FL-BMDC but not on MoDC and GM-BMDC. MoDC and FL-DC responded well to TLR ligands including poly-IC (TLR2), Pam3Cys (TLR3), LPS (TLR4) and imiquimod (TLR7) by up-regulating MHC II and CD86. The generated DC and MΦ showed a stimulatory capacity for lymphocytes, which increased upon maturation with LPS. Taken together, our results are the basis for further characterization of canine DC subsets with respect to their role in inflammation and immune responses.

Suppression of canine myeloid cells by soluble factors from cultured canine tumor cells

“…The higher CD11c expression in BM-MΦs was somewhat surprising but may be due to the fact that BM-MΦs in these studies were identified exclusively based on adherence characteristics in culture conditions that are DC-promoting (i.e. GM-CSF) but do not necessarily inhibit macrophage differentiation (Lutz et al, 1999; Ricklin Gutzwiller et al, 2010). At present, definitive generation of canine BM-MΦs is limited by a lack of recombinant canine M-CSF, a growth factor that generates macrophages.…”

“…Adherence and morphologic characteristics of DH82 cells in culture are similar to the DC cell line DC2.4, and GM-CSF treatment generated BM-DCs in mice and dogs (Lutz et al, 1999; Papenfuss, 2010, unpublished observations; Shen et al, 1997). Unlike in mice, a single individual marker such as CD11c or CD1c does not exclusively identify DC populations or differentiate these from monocytes or macrophages (Ricklin Gutzwiller et al, 2010). For our studies, we compared CD11c, CD11b and MHC class II expression between three myeloid cell populations and found that BM-DCs expressed similar CD11c, CD11b and MHC class II levels as DH82 cells, which differed from expression patterns in BM-MΦs.…”

“…For our studies, we compared CD11c, CD11b and MHC class II expression between three myeloid cell populations and found that BM-DCs expressed similar CD11c, CD11b and MHC class II levels as DH82 cells, which differed from expression patterns in BM-MΦs. Interestingly, BM-MΦs expressed higher CD11c levels than both DC populations further highlighting that CD11c cannot be exclusively used to identify canine DCs (Ricklin Gutzwiller et al, 2010). The higher CD11c expression in BM-MΦs was somewhat surprising but may be due to the fact that BM-MΦs in these studies were identified exclusively based on adherence characteristics in culture conditions that are DC-promoting (i.e.…”

Adipose tissue macrophages in non-rodent mammals: a comparative study

“…The CD206-encoding gene is conserved from birds to mammals (Staines et al 2014); however, we have little information on the macrophage expression of CD206 outside of rodents. Equine myeloid cells express CD206 (Steinbach et al 2005) and dog CD206 expression (Gene ID: 487114) has been reported in in vitro differentiated macrophages and in bone-marrow-derived dendritic cells (Ricklin Gutzwiller et al 2010). One excretory product of the canine parasite Toxocara canis, which helps evade host defense and allows survival in the tissues, has a sequence similarity to CD206 (Loukas et al 2000).…”

<b>Efeitos dos fatores tumorais derivados do melanoma canino na geração e maturação de células dendríticas caninas:</b> estudo <i>in vitro</i>

“…A redução da atividade fagocítica com o processo de maturação foi descrita por Sallusto et al (1995) Assim como os humanos e roedores, a atividade aloestimulatória em cães pode ser realizada por meio do co-cultivo das DCs com linfócitos, seguido pela medição da proliferação celular realizada por meio da incorporação do CFSE-DA aos linfócitos e avaliação da divisão celular por citometria de fluxo (WIJEWARDANA et al, 2006;WANG et al, 2007b;GUTZWILLER et al, 2010).…”