TY - JOUR A1 - Gierlich, Philipp A1 - Lex, Veronika A1 - Technau, Antje A1 - Keupp, Anne A1 - Morper, Lorenz A1 - Glunz, Amelie A1 - Sennholz, Hanno A1 - Rachor, Johannes A1 - Sauer, Sascha A1 - Marcu, Ana A1 - Grigoleit, Götz Ulrich A1 - Wölfl, Matthias A1 - Schlegel, Paul G. A1 - Eyrich, Matthias T1 - Prostaglandin E\(_2\) in a TLR3‑ and 7/8‑agonist‑based DC maturation cocktail generates mature, cytokine‑producing, migratory DCs but impairs antigen cross‑presentation to CD8\(^+\) T cells JF - Cancer Immunology, Immunotherapy N2 - Mature dendritic cells (DCs) represent cellular adjuvants for optimal antigen presentation in cancer vaccines. Recently, a combination of prostaglandin E\(_2\) (PGE\(_2\)) with Toll-like receptor agonists (TLR-P) was proposed as a new standard to generate superior cytokine-producing DCs with high migratory capacity. Here, we compare TLR-P DCs with conventional DCs matured only with the proinflammatory cytokines TNFα and IL-1ß (CDCs), focussing on the interaction of resulting DCs with CD8\(^+\) T-cells. TLR-P matured DCs showed elevated expression of activation markers such as CD80 and CD83 compared to CDCs, together with a significantly higher migration capacity. Secretion of IL-6, IL-8, IL-10, and IL-12 was highest after 16 h in TLR-P DCs, and only TLR-P DCs secreted active IL-12p70. TLR-P DCs as well as CDCs successfully primed multifunctional CD8\(^+\) T-cells from naïve precursors specific for the peptide antigens Melan-A, NLGN4X, and PTP with comparable priming efficacy and T-cell receptor avidity. CD8\(^+\) T-cells primed by TLR-P DCs showed significantly elevated expression of the integrin VLA-4 and a trend for higher T-cell numbers after expansion. In contrast, TLR-P DCs displayed a substantially reduced capability to cross-present CMVpp65 protein antigen to pp65-specific T cells, an effect that was dose-dependent on PGE2 during DC maturation and reproducible with several responder T-cell lines. In conclu-sion, TLR-P matured DCs might be optimal presenters of antigens not requiring processing such as short peptides. However, PGE\(_2\) seems less favorable for maturation of DCs intended to process and cross-present more complex vaccine antigens such as lysates, proteins or long peptides. KW - dendritic cells KW - cancer vaccines KW - prostaglandin E2 KW - TLR agonists KW - tumor-specific CD8+ T cells Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-232311 SN - 0340-7004 VL - 69 ER - TY - THES A1 - Lex, Veronika T1 - Auswirkung verschiedener Ausreifungscocktails auf die Kreuzpräsentationsfähigkeit dendritischer Zellen T1 - Effect of different maturation cocktails on cross-presentation ability of dendritic cells N2 - Ziel dieser Arbeit war die Entwicklung optimaler DCs für die Generierung von therapeutischen Tumorvakzinen. Neben den essenziellen Eigenschaften der DCs wie der Migrationsfähigkeit, der Hochregulation kostimulatorischer Moleküle sowie der Zytokinausschüttung, ist auch die optimale Aufnahme von Tumor-spezifischen Antigenen und deren Präsentation besonders wichtig, um eine effiziente Immunantwort zur ermöglichen. Wichtigstes Ziel war es über einen optimalen Ausreifungscocktail der DCS eine effektive Antwort der zytotoxischen CD8\(^+\)-Zellen zu erzielen. Dies geschieht im Rahmen der Präsentation von z.B. exogenen Antigenen wie in unserem Falle eines CMV-Proteins über den Weg der sogenannten Kreuzpräsentation. Hierzu wurde im Rahmen dieser Arbeit der in unserer Klinik etablierte zytokinbasierte Ausreifungscocktail (IL-1β, TNFα) mit einem Ausreifungscocktail, welcher Toll-like-Rezeptor-Agonisten mit PGE\(_2\) kombiniert, verglichen. Wir konnten erstmals zeigen, dass PGE\(_2\) nicht nur einen Einfluss auf die Ausreifung, Zytokinproduktion und somit Migrationsfähigkeit der DCs hat wie in der Literatur beschrieben, sondern auch auf die Kreuzpräsentationsfähigkeit exogener Antigene. Das Weglassen von PGE\(_2\) im Cocktail 2 führte zu einer signifikant besseren Kreuzpräsentationsfähigkeit. Somit lässt sich durch unsere Experimente auf eine hemmende Wirkung von PGE\(_2\) auf die T-Zell-Aktivierung über die Kreuzpräsentation schließen. Als mögliche Schlussfolgerung unserer Experimente sollte bei der Arbeit mit Antigenen, welche über Kreuzpräsentation eine T-Zell-Antwort auslösen (Proteine, Tumorlysat, long-peptides) auf die Zugabe von PGE\(_2\) im Ausreifungscocktail verzichtet werden. N2 - The aim of this work was to develop optimal dentritic cells (DCs) for the generation of therapeutic tumor vaccines. Besides the essential properties of DCs such as migration ability, upregulation of costimulatory molecules as well as cytokine release, the optimal uptake of tumor-specific antigens and their presentation is also particularly important to enable an efficient immune response. The most important goal was to achieve an effective response of cytotoxic CD8\(^+\) cells via an optimal maturation cocktail for DCs. This is done in the context of the presentation of e.g. exogenous antigens like in our case a CMV protein via cross-presentation. For this purpose, we compared the cytokine-based maturation cocktail (IL-1β, TNFα) established in our clinic with a maturation cocktail combining Toll-like receptor agonists with prostaglandin E2 (PGE\(_2\)). We were able to show for the first time that PGE\(_2\) not only has an impact on maturation, cytokine production and thus migratory ability of DCs as described in the literature, but also on the cross-presentation ability of exogenous antigens. Elimination of PGE\(_2\) in cocktail 2 resulted in significantly better cross-presentation ability. Thus, our experiments suggest an inhibitory effect of PGE\(_2\) on T-cell activation via cross-presentation. As a possible conclusion of our experiments, the addition of PGE\(_2\) in the maturation cocktail should be avoided when working with antigens that trigger a T cell response via cross-presentation (proteins, tumor lysate, long-peptides). KW - Dendritische Zelle KW - Toll-like-Rezeptoren KW - Tumor KW - Prostaglandin E2 KW - T-Lymphozyt KW - Kreuzpräsentation KW - Antigenspezifische T-Zelle KW - Ausreifungscocktail Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-253334 ER -