A combined approach to signal enhancement in fluorescence affinity biosensors and assays is reported. It is based on the compaction of specifically captured target molecules at the sensor surface followed by optical probing with a tightly confined surface plasmon (SP) field. This concept is utilized by using a thermoresponsive hydrogel (HG) binding matrix that is prepared from a terpolymer derived from poly(N-isopropylacrylamide) (pNIPAAm) and attached to a metallic sensor surface. Epi-illumination fluorescence and SP-enhanced total internal reflection fluorescence readouts of affinity binding events are performed to spatially interrogate the fluorescent signal in the direction parallel and perpendicular to the sensor surface. The pNIPAAm-based HG binding matrix is arranged in arrays of sensing spots and employed for the specific detection of human IgG antibodies against the Epstein-Barr virus (EBV). The detection is performed in diluted human plasma or with isolated human IgG by using a set of peptide ligands mapping the epitope of the EBV nuclear antigen. Alkyne-terminated peptides were covalently coupled to the pNIPAAm-based HG carrying azide moieties. Importantly, using such low-molecular-weight ligands allowed preserving the thermoresponsive properties of the pNIPAAm-based architecture, which was not possible for amine coupling of regular antibodies that have a higher molecular weight.

• MeSH
• akrylové pryskyřice chemie   MeSH
• biosenzitivní techniky metody   MeSH
• fluorescence MeSH
• hydrogely chemie   metabolismus   MeSH
• imunoglobulin G analýza   imunologie   MeSH
• infekce virem Epsteina-Barrové diagnóza   imunologie   metabolismus   virologie   MeSH
• lidé MeSH
• peptidové fragmenty imunologie   metabolismus   MeSH
• polymery chemie   MeSH
• virus Epsteinův-Barrové - jaderné antigeny imunologie   MeSH
• virus Epsteinův-Barrové imunologie   izolace a purifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Human Cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) are endowed with the ability of establishing lifelong latency in human hosts and reactivating in immunocompromised subjects, including patients suffering from ulcerative colitis (UC). We, therefore, aimed to investigate virus-specific immunity in UC patients. A cohort of 24 UC patients (14 responders and 10 refractory to therapy) and 26 control subjects was prospectively enrolled to undergo virus-specific serology (by ELISA assay) and assessment of both CD4+ and CD8+ virus-specific T-cell response (by interferon-γ enzyme-linked immunospotanalysis). In parallel, mucosal viral load was determined by quantitative real-time PCR and the values were correlated with both clinical and endoscopic indexes of activity. For statistics, the t-test, Mann-Withney test, Fisher's exact test and Spearman rank correlation test were applied; p < 0.05 was considered significant. EBV-specific CD4+ and CD8+ T-cell responses were significantly lower in UC patients compared to controls (p < 0.0001 and p = 0.0006, respectively), whereas no difference was found for HCMV-specific T-cell response. When dividing the UC group according to response to therapy, both responders and refractory UC patients showed a deficient EBV-specific CD4+ T-cell response with respect to controls (p < 0.04 and p = 0.0003, respectively). Moreover, both EBV and HCMV mucosal loads were significantly higher in refractory UC than in responders and controls (p = 0.007 and 0.003; and p = 0.02 and 0.001, respectively), and correlated with activity indexes. Steroid therapy seemed the main risk factor for triggering EBV colitis. Finally, no cases of IgM positivity were found in the study population. An impaired EBV-specific immunity was clearly evident in UC patients, mostly in those refractory to therapy. The ELISPOT assay may serve as new tool for quantifying and monitoring virus-specific T-cell immunity in UC.

• MeSH
• CD4-pozitivní T-lymfocyty metabolismus   MeSH
• CD8-pozitivní T-lymfocyty metabolismus   MeSH
• cytomegalovirové infekce farmakoterapie   imunologie   MeSH
• Cytomegalovirus imunologie   fyziologie   MeSH
• DNA virů genetika   MeSH
• infekce virem Epsteina-Barrové farmakoterapie   imunologie   MeSH
• lidé MeSH
• prospektivní studie MeSH
• steroidy škodlivé účinky   terapeutické užití   MeSH
• studie případů a kontrol MeSH
• ulcerózní kolitida farmakoterapie   imunologie   virologie   MeSH
• virová nálož MeSH
• virus Epsteinův-Barrové imunologie   fyziologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Adoptive transfer of multivirus-specific T cell lines (MVST) is an advanced tool for immunotherapy of virus infections after hematopoietic stem cell transplantation (HSCT). Their preparation includes activation of donor virus-specific T cells by the mixture of oligopeptides derived from immunodominant antigens of several most harmful viruses, i.e. human cytomegalovirus (HCMV), polyomavirus BK (BKV), Epstein-Barr virus (EBV) and adenovirus (ADV). The aim of our study was to find out whether antigenic competition may have an impact on the expansion of virus-specific T cells. MVST from several heathy blood donors were generated using a pulse of overlapping oligopeptides (PepMixes™, derived from the IE1 and pp65 CMV antigens, VP1 and LTAG BKV antigens, BZLF1 and EBNA1 proteins of EBV and hexon protein from ADV) and short time culture in the presence of IL-7 and IL-4. The amount of virus-specific T cells in MVST was measured by ELISPOT and flow cytometry after re-stimulation with individual antigens. To evaluate antigenic competition, MVST were expanded either with a complete set of antigens or with the mixture lacking some of them. MVST expanded with the antigen mixture including CMV antigens contained a lower proportion of the T cells of other antigen specificities. A similar inhibitory effect was not apparent for EBV-derived peptides. The competitive effect of CMV antigens was most pronounced in MVST from CMV-seropositive donors and was mediated by both IE1 and pp65-derived peptides. Antigenic competition did not influence the phenotype of either CMV- or BKV-specific T cells. Both T cell populations had an effector memory phenotype (CD45RO+, CD27-, CCR7-). However, CMV-specific T cells preferentially consist of CD8+ while in BKV-specific T cells, the CD4+ phenotype predominated. Modification of the MVST manufacture protocol to prevent antigenic competition may increase the efficacy of MVST in therapy of BKV infections in HSCT recipients.

• MeSH
• Adenoviridae imunologie   patogenita   MeSH
• adenovirové infekce lidí imunologie   terapie   virologie   MeSH
• aktivace lymfocytů MeSH
• antigeny virové imunologie   MeSH
• cytomegalovirové infekce imunologie   terapie   virologie   MeSH
• Cytomegalovirus imunologie   patogenita   MeSH
• fenotyp MeSH
• imunodominantní epitopy MeSH
• imunoterapie adoptivní * MeSH
• infekce onkogenními viry imunologie   terapie   virologie   MeSH
• infekce virem Epsteina-Barrové imunologie   terapie   virologie   MeSH
• interakce hostitele a patogenu MeSH
• kultivované buňky MeSH
• lidé MeSH
• polyomavirové infekce imunologie   terapie   virologie   MeSH
• T-lymfocyty imunologie   transplantace   virologie   MeSH
• transplantace hematopoetických kmenových buněk škodlivé účinky   MeSH
• virové nemoci imunologie   terapie   virologie   MeSH
• virus BK imunologie   patogenita   MeSH
• virus Epsteinův-Barrové imunologie   patogenita   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Klíčová slova
• Lipo C Askor, EpaRition, 2LSEP, 2LEPV,
• MeSH
• imunoterapie MeSH
• interleukin-10 terapeutické užití   MeSH
• lidé MeSH
• lymfocyty imunologie   účinky léků   virologie   MeSH
• roztroušená skleróza etiologie   imunologie   patofyziologie   MeSH
• virus Epsteinův-Barrové * imunologie   patogenita   MeSH
• Check Tag
• lidé MeSH

B lymfocyty hrají v patofyziologii roztroušené sklerózy (MS) významnou roli, což potvrzuje léčebný efekt protilátek zaměřených na dosažení jejich deplece. Myšlenka, že viry mohou hrát roli v rozvoji MS, má dlouhou historii a je podpořena studiemi prokazujícími souvislost mezi virem EpsteinBarrové (EBV) a nástupem choroby. Vývoj antivirotik pro léčbu MS a studium role virů v patologii onemocnění si zasluhují širší pozornost. V časopise Trends in Molecular Medicine byl publikován článek na téma souvislosti EBV infekce a MS.1 O tomto článku referujeme v následujícím textu.

• MeSH
• B-lymfocyty mikrobiologie   virologie   MeSH
• lidé MeSH
• roztroušená skleróza * etiologie   imunologie   terapie   MeSH
• virus Epsteinův-Barrové imunologie   patogenita   MeSH
• Check Tag
• lidé MeSH

• Klíčová slova
• Lipo-C-Askor, 2LSEP, 2LINFLAM, 2LEBV,
• MeSH
• B-lymfocyty imunologie   virologie   MeSH
• imunomodulace MeSH
• imunoterapie metody   MeSH
• lidé MeSH
• mikrobiota imunologie   účinky léků   MeSH
• roztroušená skleróza diagnostické zobrazování   imunologie   patofyziologie   virologie   MeSH
• virová nálož účinky léků   MeSH
• virus Epsteinův-Barrové imunologie   izolace a purifikace   MeSH
• zánět farmakoterapie   mikrobiologie   MeSH
• Check Tag
• lidé MeSH

Extended molecular characterization of HLA genes in the IHWG reference B-lymphoblastoid cell lines (B-LCLs) was one of the major goals for the 17th International HLA and Immunogenetics Workshop (IHIW). Although reference B-LCLs have been examined extensively in previous workshops complete high-resolution typing was not completed for all the classical class I and class II HLA genes. To address this, we conducted a single-blind study where select panels of B-LCL genomic DNA samples were distributed to multiple laboratories for HLA genotyping by next-generation sequencing methods. Identical cell panels comprised of 24 and 346 samples were distributed and typed by at least four laboratories in order to derive accurate consensus HLA genotypes. Overall concordance rates calculated at both 2- and 4-field allele-level resolutions ranged from 90.4% to 100%. Concordance for the class I genes ranged from 91.7 to 100%, whereas concordance for class II genes was variable; the lowest observed at HLA-DRB3 (84.2%). At the maximum allele-resolution 78 B-LCLs were defined as homozygous for all 11 loci. We identified 11 novel exon polymorphisms in the entire cell panel. A comparison of the B-LCLs NGS HLA genotypes with the HLA genotypes catalogued in the IPD-IMGT/HLA Database Cell Repository, revealed an overall allele match at 68.4%. Typing discrepancies between the two datasets were mostly due to the lower-resolution historical typing methods resulting in incomplete HLA genotypes for some samples listed in the IPD-IMGT/HLA Database Cell Repository. Our approach of multiple-laboratory NGS HLA typing of the B-LCLs has provided accurate genotyping data. The data generated by the tremendous collaborative efforts of the 17th IHIW participants is useful for updating the current cell and sequence databases and will be a valuable resource for future studies.

• MeSH
• alely MeSH
• B-lymfocyty virologie   MeSH
• exony genetika   MeSH
• genetická variace MeSH
• genetické lokusy MeSH
• genotyp MeSH
• haplotypy genetika   MeSH
• histokompatibilita - antigeny třídy I genetika   MeSH
• histokompatibilita - antigeny třídy II genetika   MeSH
• histokompatibilita MeSH
• HLA antigeny genetika   MeSH
• homozygot MeSH
• jednoduchá slepá metoda MeSH
• lidé MeSH
• sekvenční analýza DNA metody   MeSH
• správnost dat MeSH
• testování histokompatibility metody   MeSH
• transformované buněčné linie MeSH
• virová transformace buněk MeSH
• virus Epsteinův-Barrové imunologie   MeSH
• vysoce účinné nukleotidové sekvenování metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• antigeny CD274 klasifikace   škodlivé účinky   MeSH
• cirkulující nádorová DNA genetika   klasifikace   MeSH
• Helicobacter pylori patogenita   MeSH
• lidé MeSH
• mikrobiota MeSH
• mikrosatelitní nestabilita MeSH
• nádory žaludku * genetika   klasifikace   patologie   MeSH
• virus Epsteinův-Barrové imunologie   MeSH
• Check Tag
• lidé MeSH

• Klíčová slova
• transplantace "HIV-to-HIV",
• MeSH
• antivirové látky MeSH
• benzimidazoly aplikace a dávkování   terapeutické užití   MeSH
• Cytomegalovirus imunologie   patogenita   MeSH
• ganciklovir terapeutické užití   MeSH
• herpetické infekce farmakoterapie   prevence a kontrola   MeSH
• HIV patogenita   MeSH
• imunosupresivní léčba MeSH
• lidé MeSH
• lidský herpesvirus 8 patogenita   MeSH
• Polyomavirus patogenita   MeSH
• preexpoziční profylaxe MeSH
• rizikové faktory MeSH
• RNA-viry patogenita   MeSH
• transplantace orgánů * MeSH
• virové nemoci * farmakoterapie   prevence a kontrola   MeSH
• virus Epsteinův-Barrové imunologie   patogenita   MeSH
• virus zika patogenita   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

• Klíčová slova
• imunopatogeneze,
• MeSH
• Cytomegalovirus imunologie   patogenita   MeSH
• Herpesviridae imunologie   klasifikace   patogenita   MeSH
• imunitní únik fyziologie   imunologie   MeSH
• lidé MeSH
• lidský herpesvirus 1 imunologie   patogenita   MeSH
• přirozená imunita MeSH
• replikace viru fyziologie   imunologie   MeSH
• virové nemoci imunologie   patofyziologie   MeSH
• virus Epsteinův-Barrové imunologie   patogenita   MeSH
• virus spalniček imunologie   patogenita   MeSH
• viry hepatitidy imunologie   patogenita   MeSH
• viry * imunologie   klasifikace   patogenita   MeSH
• zánět * etiologie   imunologie   patofyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH