Generation and functional analysis of human TNF‐α/iNOS‐producing dendritic cells (Tip‐DC). Issue 7 (6th June 2013)

• Record Type: Journal Article
• Authors: Wilsmann‐Theis, D.; Koch, S.; Mindnich, C.; Bonness, S.; Schnautz, S.; von, D.; Bieber, T.
• Is Part Of: Allergy. Volume 68:Issue 7(2013:Jul.)
• Page Start: 890
• Page End: 898
• Publication Date: 2013-06-06
• Subjects: Allergy -- Periodicals; 616.97
• DOI: 10.1111/all.12172
• Languages: English
• ISSNs: 0105-4538

Abstract

<abstract abstract-type="main" id="all12172-abs-0001"> <title>Abstract</title> <sec id="all12172-sec-0001" sec-type="section"> <title>Background</title> <p>A unique type of CD11c<sup>pos</sup> dendritic cells (DC) is abundant in inflamed tissue, for example, in chronic inflammatory skin diseases. Due to their remarkable production of tumor necrosis factor (TNF)‐α and inducible NO synthase (iNOS), these cells have been referred to as TNF and iNOS‐producing DC (Tip‐DC). While Tip‐DC have been mainly characterized in murine models of infection, functional data about their human counterpart are lacking.</p> </sec> <sec id="all12172-sec-0002" sec-type="section"> <title>Objectives</title> <p>We sought to generate human Tip‐DC <italic>in vitro</italic> und thus provide a new model for the investigation of their phenotype and function.</p> </sec> <sec id="all12172-sec-0003" sec-type="section"> <title>Methods</title> <p>We generated human Tip‐DC from monocytic precursor cells of healthy individuals, atopic and psoriatic patients using human serum. Resting and stimulated cells were analyzed by flow cytometry, real‐time PCR, and by ELISA. INOS activity was measured by fluorometric detection of NO.</p> </sec> <sec id="all12172-sec-0004" sec-type="section"> <title>Results</title> <p>Tip‐DC closely resembled their <italic>in vivo</italic> counterparts by expressing CD11c, CD86, and CD40 while lacking CD1a, CD1c, or CD207/Langerin. Bacterial stimulation of Tip‐DC from healthy donors, atopic dermatitis, or psoriasis patients resulted in a similar increase in iNOS activity and TNF‐α production. In kinetic experiments, TNF‐α, a putative activator of Tip‐DC, could not induce NOS2. Upon bacterial stimulation, <italic>TNFA</italic>, <italic> IL6</italic>, <italic> IL12B, </italic> and <italic>IL23A </italic>mRNA appeared in a first wave, while <italic>IL12A</italic> and <italic>NOS2 </italic>mRNA were up‐regulated later on but not blocked by anti‐TNF‐α agents, implying a biphasic pro‐inflammatory response.</p> </sec> <sec id="all12172-sec-0005" sec-type="section"> <title>Conclusions</title> <p>We developed a new model for the study of human Tip‐DC and provide the first evidence of their pro‐inflammatory capacity.</p> </sec> </abstract>