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Published on: November 4, 2016
Immunophenotype and functional properties of feline dendritic cells derived from blood and bone marrow
D Bienzle1, F Reggeti, M E Clark
1Department of Pathobiology, University of Guelph, Guelph, Ont., Canada N1G 2W1. dbienzle@uoguelph.ca
Insights
Researchers successfully derived feline dendritic cells (DCs) from blood and bone marrow. These cells exhibit key markers and functions, paving the way for studying feline infectious diseases.
Area of Science:
- Immunology
- Veterinary Science
Background:
- Dendritic cells (DCs) are crucial for initiating immune responses.
- The specific identity and function of feline DCs remain largely unknown.
- Understanding feline DCs is vital for addressing infectious diseases in cats.
Purpose of the Study:
- To derive and characterize feline dendritic cells (DCs) in vitro.
- To establish a method for generating a consistent population of feline DCs.
- To investigate the surface markers and functional capabilities of these derived feline DCs.
Main Methods:
- Feline blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BMMCs) were cultured for 3-10 days.
- Interleukin-4 (IL-4) and granulocyte-macrophage colony-stimulating factor (GM-CSF) were used as cytokines.
- Surface molecule expression, phagocytic activity, and lymphocyte proliferation assays were performed.
Main Results:
- Bone marrow mononuclear cells (BMMCs) yielded higher numbers of DCs compared to PBMCs.
- Derived DCs expressed specific surface markers (CD1a, CD1b, CD1c, CD11b, CD14) and higher MHC class I and II levels than macrophages or monocytes.
- The cells demonstrated antigen uptake via phagocytosis and pinocytosis and induced allogeneic lymphocyte proliferation.
Conclusions:
- The culture method successfully generated feline cells analogous to immature myeloid dendritic cells.
- These feline DCs possess characteristic markers and functions of myeloid DCs.
- The availability of these feline DCs will facilitate research into their role in feline infectious diseases and potential therapeutic uses.
Abstract:
Dendritic cells (DCs) are a heterogeneous population of cells of fundamental importance in initiating innate as well as specific immune responses. The identity and function of DCs in the cat are unknown, although they are likely pivotal in the response to infection. In this study, feline DCs were derived by 3-10-day culture of adherent blood mononuclear cells (PBMCs) and bone marrow mononuclear cells (BMMCs) in the presence of IL 4 and GM-CSF. BMMC consistently yielded a greater number of DCs than PBMC, and there were fewer macrophages than DC from both compartments. DCs expressed a distinct constellation of surface molecules, which included CD1a, CD1b, and CD1c, CD11b, CD14, and 2-3-fold higher levels of MHC class I and II molecules than co-cultured macrophages or fresh blood monocytes. DCs displayed typical cytoplasmic processes, limited non-specific esterase activity, and acquired antigen by phagocytosis, pinocytosis, and binding to specific receptors. Cytokine-exposed cells induced proliferation of allogeneic lymphocytes. Thus, the cells derived by these culture conditions had markers and functions analogous to immature myeloid DCs. Availability of feline DCs will enable investigation of their role in infectious disease and their potential therapeutic application.
