Rheumatoid arthritis is an autoimmunity leading to considerable impairment of quality of life. N-acetyl glucosamine (GlcNAc) has been described previously as a potent modulator of experimental arthritis in animal models and is used for osteoarthritis treatment in humans, praised for its lack of adverse effects. In this study we present a comprehensive immunological analysis of multivalent GlcNAc-terminated glycoconjugate (GC) application in the treatment of collagen-induced arthritis (CIA) and its clinical outcome. We used immunohistochemistry and FACS to describe conditions on the inflammation site. Systemic and clinical effects were evaluated by FACS, cytotoxicity assay, ELISA, cytometric bead array (CBA), RT-PCR and clinical scoring. We found reduced inflammatory infiltration, NKG2D expression on NK and suppression of T, B and antigen-presenting cells (APC) in the synovia. On the systemic level, GCs prevented the activation of monocyte- and B cell-derived APCs, the rise of TNF-α and IFN-γ levels, and subsequent type II collagen (CII)-specific IgG2a formation. Moreover, we detected an increase of anti-inflammatory IL-4 mRNA in the spleen. Similar to the synovia, the GCs caused a significant reduction of NKG2D-expressing NK cells in the spleen without influencing their lytic function. GCs effectively postponed the onset of arthritic symptoms, reduced their severity and in 18% (GN8P) and 31% (GN4C) of the cases completely prevented their appearance. Our data prove that GlcNAc glycoconjugates prevent the inflammatory response, involving proinflammatory cytokine rise, APC activation and NKG2D expression, leading to the attenuation of clinical symptoms. These results support the glycobiological approach to the treatment of collagen-induced arthritis/rheumatoid arthritis (CIA/RA) as a way of bringing new prospects for more effective therapeutic interventions.

• Klíčová slova
• CIA, GlcNAc glycoconjugates, clinical scoring, cytokines, humoral response,
• MeSH
• acetylglukosamin aplikace a dávkování   MeSH
• antigen prezentující buňky účinky léků   imunologie   MeSH
• artritida experimentální farmakoterapie   imunologie   MeSH
• B-lymfocyty účinky léků   imunologie   MeSH
• buněčná diferenciace účinky léků   MeSH
• buňky NK účinky léků   imunologie   MeSH
• cytokiny metabolismus   MeSH
• kultivované buňky MeSH
• lektinové receptory NK-buněk - podrodina K metabolismus   MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední DBA MeSH
• myši MeSH
• revmatoidní artritida farmakoterapie   imunologie   MeSH
• synoviální membrána účinky léků   imunologie   MeSH
• T-lymfocyty účinky léků   imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• acetylglukosamin MeSH
• cytokiny MeSH
• Klrk1 protein, mouse MeSH Prohlížeč
• lektinové receptory NK-buněk - podrodina K MeSH

BACKGROUND: Dendritic cells (DCs) are professional antigen-presenting cells that are capable of inducing immune responses. DC-based vaccines are normally generated using a standard 5- to 7-day protocol. To shorten the DC-based vaccine production for use in cancer immunotherapy, we have developed a fast DC protocol by comparing standard DCs (Day 5 DCs) and fast DCs (Day 3 DCs). METHODS: We tested the generation of Day 5 versus Day 3 DCs using CellGro media and subsequent activation by two activation stimuli: Poly (I:C) and LPS. We evaluated DC morphology, viability, phagocyte activity, cytokine production and ability to stimulate antigen-specific T cells. RESULTS: Day 5 and Day 3 DCs exhibited similar phagocytic capacity. Poly (I:C)-activated Day 5 DCs expressed higher levels of the costimulatory and surface molecules CD80, CD86 and HLA-DR compared to Poly (I:C)-activated Day 3 DCs. Nevertheless, LPS-activated Day 5 and Day 3 DCs were phenotypically similar. Cytokine production was generally stronger when LPS was used as the maturation stimulus, and there were no significant differences between Day 5 and Day 3 DCs. Importantly, Day 5 and Day 3 DCs were able to generate comparable numbers of antigen-specific CD8(+) T cells. The number of Tregs induced by Day 5 and Day 3 DCs was also comparable. CONCLUSION: We identified monocyte-derived DCs generated in CellGro for 3 days and activated using Poly (I:C) similarly potent in most functional aspects as DCs produced by the standard 5 day protocol. These results provide the rationale for the evaluation of faster protocols for DC generation in clinical trials.

• Klíčová slova
• Cancer immunotherapy, Dendritic cell, Fast generation, Monocyte, Vaccine,
• MeSH
• antigen prezentující buňky účinky léků   imunologie   metabolismus   MeSH
• antigenní specifita receptorů T-buněk imunologie   MeSH
• buněčná diferenciace účinky léků   imunologie   MeSH
• buněčné kultury * MeSH
• časové faktory MeSH
• cytokiny biosyntéza   MeSH
• dendritické buňky cytologie   účinky léků   imunologie   metabolismus   MeSH
• fagocytóza imunologie   MeSH
• fenotyp MeSH
• imunofenotypizace MeSH
• imunoterapie MeSH
• lidé MeSH
• nádory imunologie   metabolismus   terapie   MeSH
• poly I-C imunologie   farmakologie   MeSH
• protinádorové vakcíny imunologie   MeSH
• T-lymfocyty - podskupiny imunologie   metabolismus   MeSH
• viabilita buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH
• poly I-C MeSH
• protinádorové vakcíny MeSH