BACKGROUND: The anti-PD-1 monoclonal antibody pembrolizumab has been shown to be associated with a good response in patients with metastatic gastric cancer. Excellent therapeutic results of pembrolizumab have been shown in patients with tumours showing a high microsatellite instability (MSI) and Epstein–Barr virus (EBV) positivity. GOAL: This is a retrospective study of 40 bioptic specimens from the patients, who underwent gastrectomy for gastric carcinoma. The goal of the study was to identify biomarkers (EBV, MLH-1, PDL-1 expression) that are potentially relevant for selecting the patients, who may benefit from PD-1 inhibition therapy. METHODS: Immunohistochemical (IHC) expression of PDL-1 and MSI, cytogenetic FISH amplification of the HER-2/neu gene and polymerase chain reaction of EBV RNA, including charge quantification, were performed in selected patients with metastatic or advanced gastric cancer. RESULTS: EBV-encoded RNA was detected in nine patients. None of them exhibited Her-2 overexpression or CMV infection. PD-L1 was detected in twelve patients. Ten patients were MLH1 positive. All nine cases of EBV infection showed a high expression of PD-L1 and MLH-1 (Tab. 1, Ref. 14).
- MeSH
- antigeny CD274 analýza imunologie MeSH
- biologické markery * analýza MeSH
- exprese genu MeSH
- infekce virem Epsteina-Barrové diagnóza imunologie MeSH
- klinické laboratorní techniky metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikrosatelitní nestabilita MeSH
- MutL homolog 1 analýza imunologie MeSH
- nádory žaludku * imunologie patofyziologie MeSH
- odběr biologického vzorku MeSH
- receptor erbB-2 analýza imunologie MeSH
- retrospektivní studie MeSH
- senioři MeSH
- výběr pacientů MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- senioři MeSH
- Publikační typ
- práce podpořená grantem MeSH
- MeSH
- autoimunita * MeSH
- autoimunitní nemoci etiologie patofyziologie virologie MeSH
- cytomegalovirové infekce imunologie komplikace MeSH
- hepatitida C imunologie komplikace MeSH
- herpetické infekce imunologie komplikace MeSH
- HIV infekce imunologie komplikace MeSH
- infekce virem Epsteina-Barrové imunologie komplikace MeSH
- koronavirové infekce imunologie komplikace MeSH
- lidé MeSH
- retrovirové infekce imunologie komplikace MeSH
- virové nemoci * imunologie komplikace MeSH
- Check Tag
- lidé MeSH
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
- MeSH
- autoimunitní tyreoiditida * diagnóza etiologie farmakoterapie MeSH
- depresivní poruchy imunologie komplikace MeSH
- dospělí MeSH
- imunoterapie metody MeSH
- infekce virem Epsteina-Barrové diagnóza imunologie komplikace MeSH
- lidé MeSH
- probiotika * terapeutické užití MeSH
- virové nemoci imunologie komplikace MeSH
- výsledek terapie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- Klíčová slova
- postcovidový syndrom,
- MeSH
- COVID-19 * imunologie komplikace MeSH
- infekce virem Epsteina-Barrové * imunologie komplikace MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- pozorovací studie MeSH
AIMS: Epstein-Barr virus (EBV) targets predominantly B cells and these cells could acquire new phenotype characteristics. Here we analyzed whether EBV-infected and -uninfected B cells from healthy subjects differ in proportion of dominant phenotypes, maturation stage, and homing receptors expression. METHODS: EBV-infected and -uninfected cells were identified by flow cytometry using fluorophore-labeled EBV RNA-specific DNA probes combined with fluorophore-labeled antibody to surface lineage markers, integrins, chemokine receptors, and immunoglobulin isotypes, including intracellular ones. RESULTS: Our results show that the trafficking characteristics of EBERpos B cells are distinct from EBERneg B cells with most dominant differences detected for α4β1 and α4β7 and CCR5 and CCR7. EBV-positive cells are predominantly memory IgM+ B cells or plasmablasts/plasma cells (PB/PC) positive for IgA or less for IgM. In comparison to uninfected B cells, less EBV-positive B cells express α4β7 and almost no cells express α4β1. EBV-positive B cells contained significantly higher proportion of CCR5+ and CCR7+ cells in comparison to EBV-negative cells. In vitro exposure of blood mononuclear cells to pro-inflammatory cytokine IL-6 reduces population of EBV-positive B cell. CONCLUSION: Although EBV-infected B cells represent only a minor subpopulation, their atypical functions could contribute in predisposed person to development abnormities such as some autoimmune diseases or tumors. Using multi-parameter flow cytometry we characterized differences in migration of EBV-positive and -negative B cells of various maturation stage and isotype of produced antibodies particularly different targeting to mucosal tissues of gastrointestinal and respiratory tracts.
- MeSH
- B-lymfocyty imunologie MeSH
- dospělí MeSH
- infekce virem Epsteina-Barrové imunologie patofyziologie MeSH
- krev imunologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- průtoková cytometrie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- vezikulární transportní proteiny imunologie metabolismus MeSH
- zdraví dobrovolníci pro lékařské studie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie 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
The role of human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) in the exacerbation of inflammatory bowel disease (IBD) is still uncertain. We prospectively investigated the presence of EBV and HCMV infection in both epithelial and immune cells of colonic mucosa of IBD patients, both refractory and responders to standard therapies, in comparison with patients suffering from irritable bowel syndrome who were considered as controls, by using quantitative real-time polymerase chain reaction, immunohistochemistry and in situ hybridization, in an attempt to assess viral localization, DNA load, life cycle phase and possible correlation with disease activity indexes. We obtained clear evidence of the presence of high DNA loads of both viruses in either enterocytes or immune cells of refractory IBD patients, whereas we observed low levels in the responder group and an absence of detectable copies in all cell populations of controls. Remarkably, the values of EBV and HCMV DNA in inflamed mucosa were invariably higher than in non-inflamed areas in both IBD groups, and the EBV DNA loads in the cell populations of diseased mucosa of refractory IBD patients positively correlated with the severity of mucosal damage and clinical indexes of activity. Moreover, EBV infection resulted the most prevalent either alone or in combination with HCMV, while immunohistochemistry and in situ hybridization did not allow us to distinguish between the different phases of viral life cycle. Finally, as regards treatment, these novel findings could pave the way for the use of new antiviral molecules in the treatment of this condition.
- MeSH
- cytomegalovirové infekce imunologie MeSH
- Cytomegalovirus fyziologie MeSH
- dospělí MeSH
- idiopatické střevní záněty imunologie virologie MeSH
- infekce virem Epsteina-Barrové imunologie MeSH
- kolon imunologie virologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- progrese nemoci MeSH
- prospektivní studie MeSH
- střevní sliznice imunologie virologie MeSH
- virus Epsteinův-Barrové fyziologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
