Enhanced extracellular transfer of HLA-DQ activates CD3+ lymphocytes towards compromised Treg induction in celiac disease

Celiac disease (CeD) manifests with autoimmune intestinal inflammation from gluten and genetic predisposition linked to human leukocyte antigen class-II (HLA-II) gene variants. Antigen presenting cells facilitate gluten exposition through the interaction of their surface major histocom-patibility complex (MHC) with the T cell receptor (TCR) on T lymphocytes. This fundamental mechanism of adaptive immunity has broadened upon recognition of extracellular exosomal MHC, raising awareness of an alternative means for antigen presentation. This study demonstrates that conditioned growth media (CGM) previously exposed to monocyte-derived dendritic cells from CeD significantly downregulates the CD3+ lineage marker of control T cells. Such increased activation was reflected in their elevated IL-2 secretion. Exosome localization motif identification and quantification within HLA-DQA1 and HLA DQB1 transcripts highlighted their significant prevalence within HLA-DQB1 alleles associated with CeD susceptibility. Flow cytometry revealed the strong correlation between HLA-DQ and the CD63 exosomal marker in T cells exposed to CGM from MoDCs sourced from CeD patients. This resulted in lower concentrations of CD25+ CD127− T cells, suggestive of their compromised induction to T-regulatory cells associated with CeD homeostasis. This foremost comparative study deciphered the genomic basis and extracellular exosomal effects of HLA transfer on T lymphocytes in the context of CeD, offering greater insight into this auto-immune disease

Analýza a možnosti zvýšení konverzního poměru elektronického obchodu prohifi.cz

Tato bakalářská práce je zaměřená na ukazatel úspěšnosti internetových obchodů a jeho vyhodnocování, konverzní poměr. Teoretická část přibližuje konverze jako takové, konverzní poměr, kroky obecně vedoucí k jeho zvýšení a také jeden z mnoha nástrojů, kterým lze konverzní poměr měřit. V aplikační části je analyzován konkrétní elektronický obchod prohifi.cz

Implementation of the business competency model

The thesis focuses on the implementation of the Business Competency Model in the retail chain, whose sole owner is the German parent company. In the theoretical part are - based on literature research - defined the basic concepts needed for further understanding of the thesis. The practical part first explains the cultural differences between Czechs and Germans, later the findings are used to design a competency model for the Czech Republic. The methods of semi structured interviews and pilot certification are used. In the chapters 5 and 6 are described the steps of the original German model, which is later adapted to conditions in the Czech Republic. Definitions of risk that may arise during the phases of the project and possible ways for their elimination are also considered. The thesis concludes with recommendations for the use of models in the future

Analýza a možnosti zvýšení konverzního poměru elektronického obchodu prohifi.cz

Tato bakalářská práce je zaměřená na ukazatel úspěšnosti internetových obchodů a jeho vyhodnocování, konverzní poměr. Teoretická část přibližuje konverze jako takové, konverzní poměr, kroky obecně vedoucí k jeho zvýšení a také jeden z mnoha nástrojů, kterým lze konverzní poměr měřit. V aplikační části je analyzován konkrétní elektronický obchod prohifi.cz

Pimecrolimus Is a Potent Inhibitor of Allergic Reactions to Hymenopteran Venom Extracts and Birch Pollen Allergen In Vitro.

Pimecrolimus (Elidel, SDZ ASM 981) is an anti-inflammatory and immunomodulatory 33-epichloro-derivative of macrolactam ascomycin, with low potential for affecting systemic immune responses compared with other calcineurin inhibitors, cyclosporin A and tacrolimus. Despite numerous studies focused on the mechanism of pimecrolimus action on mast cells, only the single report has addressed pimecrolimus effects on other typical FcεRI-expressing cells, the basophils. Patients allergic to birch pollen (n = 20), hymenopteran venoms (n = 23) and 10 non-allergic volunteers were examined. Primary human basophils pre-treated or not with 0.5-50 μMol pimecrolimus were exposed to various concentrations of recombinant Bet v 1a allergen, bee or wasp venom extracts and anti-IgE for 20 min, and then examined for the expression of CD45, CD193, CD203c, CD63 and CD164 using flow cytometry. The externalization of basophil activation markers (CD63 and CD164) was equally inhibited through pimecrolimus in cells activated by recombinant pollen allergen, hymenopteran venom extracts and anti-IgE. Although the individual response rate was subject to strong variation, importantly, pre-treatment with pimecrolimus lowered the number of activated basophils in response to any of the stimuli in the basophils from all patients. The inhibition was concentration-dependent; approximately half of the basophils were inhibited in the presence of 2.5 mMol pimecrolimus. Pimecrolimus is a valuable new tool for the inhibition of hyper-reactive basophils in patients with pollen allergy and a history of anaphylactic reactions to bee or wasp venoms. Further research should address short-term use of pimecrolimus in vivo in a wide spectrum of allergic diseases

Basophil identification and activation markers used in this study.

<p>Basophil identification and activation markers used in this study.</p

Inhibition of CD63 externalization on the surface of primary human basophils activated (A) or not (B) following treatment with 0.5–50 μMol pimecrolimus.

<p>(<b>A</b>) Basophils pre-treated with various concentrations of pimecrolimus were activated for 20 min with anti-IgE (n = 1 subject), bee venom (n = 3 subjects) or wasp venom (n = 3 subjects). Linear correlation coefficient r = -0.696, p < 0.01; Spearman’s D = 1027, p < 0.01. (<b>B</b>) The basophils were only pre-treated with pimecrolimus but were not activated by any external stimulus. Linear correlation coefficient r = -0.074, p > 0.05; Spearman’s D = 514, p > 0.05.</p

Flowchart diagram of the flow cytometry data analysis.

<p>Flowchart diagram of the flow cytometry data analysis.</p

Experimental protocol used for the ex vivo analysis of the pimecrolimus action on basophil activation.

<p>Experimental protocol used for the ex vivo analysis of the pimecrolimus action on basophil activation.</p

Effects of pimecrolimus treatment on the externalization of CD63 and CD164.

<p>The frequency [%] of CD63⁺CD164⁺ cells among the total basophils (gated as SSC^lowCD193⁺CD203c⁺) is shown. The IL-3-primed basophils were pre-treated or not with 2.5 μMol pimecrolimus and analyzed prior to (<b>A</b>) and after activation with anti-IgE (<b>B</b>), birch pollen allergen Bet v 1a at 100 ng*ml^-1 (<b>C</b>) and 10 ng*ml^-1 (<b>D</b>), and hymenopteran venom at 1.0 μg*ml^-1 (<b>E</b>) and 0.1 μg*ml^-1 (<b>F</b>). The Tukey box plots are shown, the mean values are presented as lines, and the 5^th and 95^th percentiles are displayed as symbols. The significance was examined by the paired t-tests with Bonferroni correction at n = 18 and is indicated as *** (p_corr<0.001 = p_uncorr<0.000056), ** (p_corr<0.01 = p_uncorr<0.00056), * (p_corr<0.05 = p_uncorr<0.0028) and n.s. (p_corr>0.05 = p_uncorr<0.0028).</p