Profiling of primary peripheral blood‐ and monocyte‐derived dendritic cells using monoclonal antibodies from the HLDA10 Workshop in Wollongong, Australia. Issue 11 (13th November 2015)

• Record Type: Journal Article
• Authors: Autenrieth, Stella E; Grimm, Sabrina; Rittig, Susanne Malaika; Grünebach, Frank; Gouttefangeas, Cécile; Bühring, Hans‐Jörg
• Is Part Of: Clinical & translational immunology. Volume 4:Issue 11 (2015)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2015-11-13
• Subjects: Immunologic diseases -- Periodicals; Immunology -- Periodicals; Clinical medicine -- Periodicals; Immune System Diseases -- therapy; Immunotherapy; Immunologic Factors -- therapeutic use; Translational Medical Research; Molecular Targeted Therapy; Clinical medicine; Immunologic diseases; Immunology; Periodicals; Periodicals; Fulltext; Internet Resources; Periodicals; 616.079
• DOI: 10.1038/cti.2015.29
• Languages: English
• ISSNs: 2050-0068

Abstract

Abstract : Dendritic cells (DCs) arise from hematopoietic stem cells and develop into a discrete cellular lineage distinct from other leucocytes. Mainly three phenotypically and functionally distinct DC subsets are described in the human peripheral blood (PB): plasmacytoid DCs (pDCs), which express the key marker CD303 (BDCA‐2), and two myeloid DC subsets (CD1c + DC (mDC1) and CD141 + DC (mDC2)), which express the key markers CD1c (BDCA‐1) and CD141 (BDCA‐3), respectively. In addition to these primary cell subsets, DCs can also be generated in vitro from either CD34 + stem/progenitor cells in the presence of Flt3 (Fms‐related tyrosine kinase 3) ligand or from CD14 + monocytes (monocyte‐derived DCs (mo‐DCs)) in the presence of granulocyte–macrophage colony‐stimulating factor+interleukin‐4 (GM‐CSF+IL‐4). Here we compare the reactivity patterns of HLDA10 antibodies (monoclonal antibody (mAb)) with pDCs, CD1c + DCs and CD141 + DCs, as well as with CD14 + ‐derived mo‐DCs cultured for 7 days in the presence of 100 ng/ml GM‐CSF plus 20 ng/ml IL‐4. A detailed profiling of these DC subsets based on immunophenotyping and multicolour flow cytometry analysis is presented. Using the panel of HLDA10 Workshop mAb, we could verify known targets selectively expressed on discrete DC subsets including CD370 as a selective marker for CD141 + DCs and CD366 as a marker for both myeloid subsets. In addition, vimentin and other markers are heterogeneously expressed on all three subsets, suggesting the existence of so far not identified DC subsets. Cell biology: Testing tools for immune cell 'fingerprinting': A diverse arsenal of antibodies reveals protein markers for various kinds of immune cells, and hints at yet unidentified cellular subtypes. Immunologists have identified three major subtypes of dendritic cells, which help 'train' the body's immune system to recognize both friend and foe. Researchers led by Stella Autenrieth and Hans‐Jörg Bühring of the University Clinic of Tübingen, Germany, have tested a collection of antibodies against diverse cell surface proteins in order to identify which proteins are found on the different dendritic cell subtypes. Their results were generally consistent with prior findings, although the quality of the individual antibodies varied. They also identified at least one marker for which the dendritic cell expression pattern was notably different than expected. Interestingly, three antibodies also generated highly variable labeling patterns within individual dendritic cell subtypes, indicating that these categories may conceal unexpected heterogeneity.