Východiska: Erdheim‑Chesterova nemoc (Erdheim‑Chester disease – ECD) je vzácná histiocytóza tzv. „nelangerhansových buněk“ spojená s intenzivní aktivací imunitního systému. Na našem klinickém pracovišti byl ECD pacient léčen anakinrou. Anakinra je antagonista receptoru pro interleukin‑1 (interleukin‑1 receptor antagonist – IL‑1RA). Tato léčba vedla ke zlepšení klinického stavu a výraznému snížení patologické únavy provázející toto onemocnění. Cílem této práce bylo analyzovat změny cytokinových profilů a změnu v zastoupení buněk imunitního systému pomocí průtokové cytometrie u ECD pacienta na počátku léčby anakinrou i po jejím skončení a srovnat je s profily zdravých dárců. Metody: Singleplex reakce 19 jednotlivých cytokinů ze séra ECD pacienta byly měřeny pomocí metody FACS array. Dále byla provedena analýza buněk z periferní krve pomocí průtokové cytometrie. Výsledky: Nejzajímavějším výsledkem bylo výrazné snížení hladiny IL‑6 bezprostředně po zahájení léčby anakinrou. Tyto výsledky naznačují významnou roli dráhy IL‑1 v patofyziologii ECD. Pomocí průtokové cytometrie jsme zaznamenali významné zvýšení počtu CD16+ monocytů před zahájením léčby, což je zcela nový poznatek. Závěr: Naše výsledky naznačují, že IL‑6 by se mohl stát markerem časné odpovědi na léčbu u ECD pacientů léčených anakinrou. Klíčová slova: Erdheim‑Chesterova nemoc – anakinra – cytokiny – průtoková cytometrie – FACS array Práce byla podpořena grantem MŠMT ČR MSM 0021622434, granty IGA MZ ČR NT14575, NT12130, grantem MZ ČR – RVO (FNBr, 65269705) a grantem Masarykovy univerzity MUNI/A/0723/2012. Autoři deklarují, že v souvislosti s předmětem studie nemají žádné komerční zájmy. Redakční rada potvrzuje, že rukopis práce splnil ICMJE kritéria pro publikace zasílané do biomedicínských časopisů. Obdrženo: 21. 2. 2014 Přijato: 10. 4. 2014

Background: Erdheim‑Chester disease (ECD) is a rare non‑Langerhans cells histiocytosis associated with intense immune activation. In our clinical center, an ECD patient was treated with anakinra, IL‑1RA (interleukin‑1 receptor antagonist), resulting in clinical improvement and major decrease of pathological fatigue. The aim of the study was to evaluate changes in cytokine profile and shift of immune cells estimated by flow cytometric analysis of ECD patient before, during initial stages of anakinra treatment as well as after treatment ceased in comparison to healthy donors. Methods: Singleplex reactions of 19 individual cytokines from serum of ECD patient were measured by FACS array. Flow cytometric analyses were performed on peripheral blood cells. Results: The most striking result is substantial decrease of IL‑6 immediately after anakinra treatment started suggesting a major role of IL‑1 pathway in ECD pathophysiology. As for flow cytometric analysis, increased number of CD16+ monocytes before treatment is a new finding. Conclusion: Our results suggest that IL‑6 may be a marker of early treatment response of ECD patients treated with anakinra.

• Keywords
• FACS array,
• MeSH
• Interleukin 1 Receptor Antagonist Protein * administration & dosage   MeSH
• C-Reactive Protein MeSH
• Cytokines * immunology   blood   MeSH
• Erdheim-Chester Disease * drug therapy   immunology   blood   MeSH
• Leukocytes cytology   drug effects   MeSH
• Humans MeSH
• Monocytes cytology   drug effects   MeSH
• Statistics, Nonparametric MeSH
• Flow Cytometry statistics & numerical data   MeSH
• Case-Control Studies MeSH
• Fatigue drug therapy   complications   MeSH
• Treatment Outcome MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Research Support, Non-U.S. Gov't MeSH

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.

• MeSH
• Acetylglucosamine administration & dosage   MeSH
• Antigen-Presenting Cells drug effects   immunology   MeSH
• Arthritis, Experimental drug therapy   immunology   MeSH
• B-Lymphocytes drug effects   immunology   MeSH
• Cell Differentiation drug effects   MeSH
• Killer Cells, Natural drug effects   immunology   MeSH
• Cytokines metabolism   MeSH
• Cells, Cultured MeSH
• NK Cell Lectin-Like Receptor Subfamily K metabolism   MeSH
• Humans MeSH
• Disease Models, Animal MeSH
• Mice, Inbred DBA MeSH
• Mice MeSH
• Arthritis, Rheumatoid drug therapy   immunology   MeSH
• Synovial Membrane drug effects   immunology   MeSH
• T-Lymphocytes drug effects   immunology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Vav3 is a key modulator of GTP-hydrolases of the Rho/Rac family, which are crucially involved in cell proliferation. Vav3 is alternatively spliced in full-length Vav3-alpha and N-terminal truncated Vav3.1 lacking its self-regulatory domains. The aim of our study was to estimate the clinical impact of Vav3 and all other Vav family members in ovarian cancer. Purification of a stem-cell like side-population (SP) from ovarian cancer cell lines was performed by flow cytometry/FACS. Differences in gene expression between SP and NSP were assessed by Gene Array analysis and confirmed by RT-PCR and immunoblot. In addition, Vav mRNA expression was determined in 150 epithelial ovarian cancers. Clinicopathological parameters, platinum-sensitivity and survival were analyzed and associated with Vav expression. SP fractions of ovarian cancer cell lines exhibited marked overexpression of Vav3.1 (p < 0.001). Vav1 and Vav2 did not prove to be of clinicopathologic relevance in ovarian cancer. High Vav3.1 expression correlated with higher FIGO stage and residual disease. Furthermore, Vav3.1 overexpression was associated with poor progression-free (HR = 2.820, p = 0.0001) and overall survival (HR = 2.842, p = 0.0001). Subgroup analyses revealed an impact of Vav3.1 on survival in Type-II but not in Type-I cancers. Notably, platinum-refractory cancers showed marked overexpression of Vav3.1 compared to other subsets of platinum-sensitivity (15.848 vs. 6.653, p = 0.0001). In conclusion, Vav3.1 is over-expressed in stem-cell like SP fractions and is clinically relevant in the pathophysiology of ovarian cancer. The N-terminal truncated Vav3.1 may be decisively involved in mechanisms causing genuine multi-drug resistance.

• MeSH
• Phosphorylation drug effects   MeSH
• Middle Aged MeSH
• Humans MeSH
• RNA, Messenger metabolism   MeSH
• Cell Line, Tumor MeSH
• Neoplastic Stem Cells drug effects   metabolism   MeSH
• Ovarian Neoplasms drug therapy   genetics   metabolism   MeSH
• Organoplatinum Compounds therapeutic use   MeSH
• Disease-Free Survival MeSH
• Prognosis MeSH
• Protein Isoforms metabolism   MeSH
• Proto-Oncogene Proteins c-vav metabolism   MeSH
• Gene Expression Regulation, Neoplastic drug effects   MeSH
• Aged MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH

• MeSH
• Immunity MeSH
• Immune System physiology   physiopathology   MeSH
• Immunotherapy MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• alergologie a imunologie
• biologie
• fyziologie
• NML Publication type
• kolektivní monografie

• MeSH
• Immunity MeSH
• Immune System MeSH
• Immunotherapy MeSH
• Publication type
• Monograph MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• alergologie a imunologie
• biologie