Self-non-self discrimination is central to T cell-mediated immunity. The kinetic proofreading model can explain T cell antigen receptor (TCR) ligand discrimination; however, the rate-limiting steps have not been identified. Here, we show that tyrosine phosphorylation of the T cell adapter protein LAT at position Y132 is a critical kinetic bottleneck for ligand discrimination. LAT phosphorylation at Y132, mediated by the kinase ZAP-70, leads to the recruitment and activation of phospholipase C-γ1 (PLC-γ1), an important effector molecule for T cell activation. The slow phosphorylation of Y132, relative to other phosphosites on LAT, is governed by a preceding glycine residue (G131) but can be accelerated by substituting this glycine with aspartate or glutamate. Acceleration of Y132 phosphorylation increases the speed and magnitude of PLC-γ1 activation and enhances T cell sensitivity to weaker stimuli, including weak agonists and self-peptides. These observations suggest that the slow phosphorylation of Y132 acts as a proofreading step to facilitate T cell ligand discrimination.

• MeSH
• adaptorové proteiny signální transdukční imunologie   metabolismus   MeSH
• aktivace lymfocytů * MeSH
• fosfolipasa C gama metabolismus   MeSH
• fosforylace imunologie   MeSH
• ligandy MeSH
• membránové proteiny imunologie   metabolismus   MeSH
• myši MeSH
• protein-tyrosinkináza ZAP-70 metabolismus   MeSH
• receptory antigenů T-buněk imunologie   metabolismus   MeSH
• T-lymfocyty imunologie   metabolismus   MeSH
• tyrosin metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

• Klíčová slova
• antineuronální protilátky, CASPR2, LGI1, nepřímá imunohistochemie,
• MeSH
• antigeny povrchové imunologie   MeSH
• autoprotilátky * imunologie   krev   MeSH
• encefalitida s protilátkami proti NMDA receptorům * diagnóza   imunologie   patofyziologie   MeSH
• imunohistochemie metody   MeSH
• imunologické techniky metody   MeSH
• lidé MeSH
• limbická encefalitida * diagnóza   imunologie   patofyziologie   MeSH
• membránové proteiny imunologie   MeSH
• paraneoplastické neurologické syndromy diagnóza   imunologie   MeSH
• proteiny nervové tkáně imunologie   MeSH
• receptory GABA-B imunologie   MeSH
• receptory N-methyl-D-aspartátu imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

INTRODUCTION: Lung cancer is the leading cause of cancer mortality worldwide; therefore, understanding the biological or clinical role of tumor-associated antigens and autoantibodies is of eminent interest for designing antitumor immunotherapeutic strategies. METHODS: Here we prospectively analyzed the serum frequencies of New York esophageal squamous cell carcinoma 1 (NY-ESO-1), human epidermal growth factor 2/neu, and melanoma-associated antigen A4 (MAGE-A4) antibodies and expression of the corresponding antigens in tumors of 121 patients with NSCLC undergoing an operation without prior neoadjuvant chemotherapy and compared them with those in 57 control age-matched patients with no history of a malignant disease. RESULTS: We found that only antibodies specific for NY-ESO-1 (19.8% [n = 24 of 121]) were significantly increased in the group of patients with NSCLC compared with in the controls. NY-ESO-1 seropositivity was significantly positively associated with an active smoking history in patients with NSCLC but not in smokers from the control group. In tumors, the frequency of NY-ESO-1 mRNA expression was 6.3% (in four of 64 patients), the frequency of human epidermal growth factor 2/neu (HER 2/neu) expression was 11.9% (five of 42), and the frequency of MAGE-A4 expression was 35.1% (20 of 57). MAGE-A4 expression in tumors correlated with smoking status and male sex in patients with NSCLC. Patients with squamous cell carcinoma displayed higher expression of NY-ESO-1 and MAGE-A4 in tumors than did patients with adenocarcinoma. On the other hand, 94.7% of nonsmoking patients in our study had adenocarcinoma (of whom 73.7% were women). CONCLUSION: These results confirm the reported high immunogenicity of NY-ESO-1 and suggest that a smoking-induced chronic inflammatory state might potentiate the development of NY-ESO-1-specific immune responses. Moreover, smoking might contribute to the expression of other cancer/testis antigens such as MAGE-A4 at early stages of NSCLC development.

• MeSH
• adenokarcinom krev   etiologie   patologie   MeSH
• antigeny nádorové krev   imunologie   MeSH
• dospělí MeSH
• kouření škodlivé účinky   MeSH
• lidé středního věku MeSH
• lidé MeSH
• membránové proteiny krev   imunologie   MeSH
• nádorové biomarkery krev   MeSH
• nádorové proteiny krev   MeSH
• nádory plic krev   etiologie   patologie   MeSH
• následné studie MeSH
• nemalobuněčný karcinom plic krev   etiologie   patologie   MeSH
• prognóza MeSH
• prospektivní studie MeSH
• receptor erbB-2 krev   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• spinocelulární karcinom krev   etiologie   patologie   MeSH
• staging nádorů MeSH
• studie případů a kontrol 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
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Personalized peptide vaccination is a promising immunotherapeutic approach in prostate cancer (PCa). We therefore examined whether an approach, utilizing personalized multiple peptide-mediated ex vivo enrichment with effector T cells reactive to multiple tumor-associated antigens (TAAs), could be employed as a basis for the development of T cell immunotherapy of PCa. In this study, we used the non-adherent fraction (lymphocytes) of cryopreserved peripheral blood mononuclear cells from a leukapheretic product of biochemically recurrent (BR, n = 14) and metastatic hormone-refractory (HR, n = 12) PCa patients. The lymphocytes were primed with a pool of mixed overlapping peptides derived from 6 PCa TAAs-PSA, PAP, NY-ESO-1, MAGE-A1, MAGE-A3 and MAGE-A4. After 2 weeks of culture, the cells were stimulated with the peptides and T cell reactivity determined by externalization of CD107a. No TAAs-reactive effector T cells were detected in the patient's lymphocytes after their reconstitution. However, following their priming with the TAAs-derived peptides and 2-week culturing, the lymphocytes became enriched with polyclonal TAAs-reactive effector CD8+ T cells in 8 out of 14 BR and 5 out of 12 HR patients. No such reactive CD8+ T cells were detected in cultured lymphocytes without the peptide priming. Stimulation of the responding cultures with peptides derived from individual TAAs revealed a unique repertoire of the reactive CD8+ T cells. Our strategy revealed that the personalized multiple peptide-mediated ex vivo enrichment with multiple TAAs-reactive T cells in the PCa patient's lymphocytes is a viable approach for development of T cell immunotherapy of PCa.

• MeSH
• antigeny nádorové imunologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• imunoterapie metody   MeSH
• individualizovaná medicína metody   MeSH
• kalikreiny imunologie   MeSH
• kultivované buňky MeSH
• leukocyty mononukleární imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• melanomové antigeny imunologie   MeSH
• membránové proteiny imunologie   MeSH
• nádorové proteiny imunologie   MeSH
• nádory prostaty imunologie   patologie   MeSH
• peptidy imunologie   MeSH
• prostatický specifický antigen imunologie   MeSH
• senioři MeSH
• T-lymfocyty imunologie   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH

Single-nucleotide polymorphism studies have linked the chromosome 17q12-q21 region, where the human orosomucoid-like (ORMDL)3 gene is localized, to the risk of asthma and several other inflammatory diseases. Although mast cells are involved in the development of these diseases, the contribution of ORMDL3 to the mast cell physiology is unknown. In this study, we examined the role of ORMDL3 in antigen-induced activation of murine mast cells with reduced or enhanced ORMDL3 expression. Our data show that in antigen-activated mast cells, reduced expression of the ORMDL3 protein had no effect on degranulation and calcium response, but significantly enhanced phosphorylation of AKT kinase at Ser 473 followed by enhanced phosphorylation and degradation of IκBα and translocation of the NF-κB p65 subunit into the nucleus. These events were associated with an increased expression of proinflammatory cytokines (TNF-α, IL-6, and IL-13), chemokines (CCL3 and CCL4), and cyclooxygenase-2 dependent synthesis of prostaglandin D2. Antigen-mediated chemotaxis was also enhanced in ORMDL3-deficient cells, whereas spreading on fibronectin was decreased. On the other hand, increased expression of ORMDL3 had no significant effect on the studied signaling events, except for reduced antigen-mediated chemotaxis. These data were corroborated by increased IgE-antigen-dependent passive cutaneous anaphylaxis in mice with locally silenced ORMDL3 using short interfering RNAs. Our data also show that antigen triggers suppression of ORMDL3 expression in the mast cells. In summary, we provide evidence that downregulation of ORMDL3 expression in mast cells enhances AKT and NF-κB-directed signaling pathways and chemotaxis and contributes to the development of mast cell-mediated local inflammation in vivo.

• MeSH
• chemotaxe * MeSH
• cytokiny genetika   imunologie   MeSH
• degranulace buněk * MeSH
• down regulace MeSH
• kultivované buňky MeSH
• mastocyty cytologie   imunologie   metabolismus   MeSH
• membránové proteiny genetika   imunologie   MeSH
• messenger RNA genetika   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• receptory IgE imunologie   MeSH
• upregulace MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

To examine evidence of positive antibodies against immunogenic proteins of Anaplasma phagocytophilum in patients with other tick-borne infections and to diagnose possible co-infections, 412 serum specimens were tested by immunoblotting using three specific Anaplasma antigens: surface proteins p44 and Asp62 and outer membrane protein A (OmpA). In total, 284 serum samples from children with Lyme borreliosis and 12 serum samples from children with tick-borne encephalitis were tested. Sera from patients with viral aseptic meningitis (n = 47) and from blood donors (n = 69) were used as controls. Among all serum specimens from patients with tick-borne infections submitted for this study, six samples (2·0%) showed positive IgM reactions and seven samples (2·4%) were IgG positive for A. phagocytophilum by immunoblot. Borderline reactivity was found in 30 samples (10·14%) for IgM and 36 samples (12·2%) for IgG. The difference between patients and blood donors was statistically significant for IgM (P = 0·006) and for IgG (P = 0·0007) antibodies. A statistically significant result was obtained for IgG (P = 0·02) but not for IgM between patients and children with aseptic meningitis. Immunoblot using three specific antigens provides novel information about the positivity of antibodies to A. phagocytophilum in children with other tick-borne infections. Taking into account clinical and laboratory findings of children despite antibody positivity, no case of human granulocytic anaplasmosis was demonstrated.

• MeSH
• Anaplasma phagocytophilum imunologie   MeSH
• anaplasmóza diagnóza   imunologie   mikrobiologie   MeSH
• bakteriální proteiny imunologie   MeSH
• dítě MeSH
• klíšťová encefalitida imunologie   mikrobiologie   MeSH
• koinfekce diagnóza   imunologie   mikrobiologie   MeSH
• lidé MeSH
• lymeská nemoc imunologie   mikrobiologie   MeSH
• membránové proteiny imunologie   MeSH
• mladiství MeSH
• předškolní dítě MeSH
• proteiny vnější bakteriální membrány imunologie   MeSH
• protilátky bakteriální krev   MeSH
• western blotting MeSH
• Check Tag
• dítě MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

• MeSH
• cílená molekulární terapie MeSH
• humanizované monoklonální protilátky aplikace a dávkování   MeSH
• imunokonjugáty aplikace a dávkování   škodlivé účinky   terapeutické užití   MeSH
• lidé MeSH
• malobuněčný karcinom plic farmakoterapie   patologie   MeSH
• membránové proteiny imunologie   účinky léků   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• zprávy MeSH

• MeSH
• analýza přežití MeSH
• cílená molekulární terapie MeSH
• humanizované monoklonální protilátky aplikace a dávkování   MeSH
• imunokonjugáty terapeutické užití   MeSH
• lidé MeSH
• malobuněčný karcinom plic farmakoterapie   patologie   MeSH
• membránové proteiny imunologie   MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• abstrakty MeSH

Imunitní systém musí být na jedné straně schopen efektivně zasáhnout proti cizím a nebezpečným patogenům, na straně druhé musí být schopen rozpoznat a tolerovat naše vlastní tkáně a orgány. Aktivita imunitního systému je ovlivňována celou řadou pozitivních (stimulačních) a negativních (inhibičních) signálů. některé z těchto inhibičních receptorů zabraňují poškození našich tkání v místě zánětu tím, že tlumí příliš silnou či dlouhou imunitní reakci. Plní tak fyziologickou ochrannou funkci před silnou zánětlivou reakcí a možnou autoimunitní patologií. některé z těchto mechanizmů jsou ovšem využívány nádory k tomu, aby unikly pozornosti imunitního systému. Další únikové strategie spočívají v produkci cytokinů a faktorů vytvářejících v nádorovém mikroprostředí silnou imunosupresi, která zabraňuje efektivní imunitní odpovědi. Tato práce si klade za cíl popsat nejčastější strategie, které jsou nádory využívány k potlačení imunitní reakce.

Immune system must be able to protect us from foreign dangerous pathogens, but on the other side, it must be able to recognize our own tissues and organs. Activity of the immune system is affected by many positive (stimulatory) and negative (inhibitory) signals. Some of these negative receptors protect us from damage of our tissues at a place of inflammation as it blocks too intensive or long‑lasting immune reaction. Thereby, they have a physiological protective function against strong inflammatory reaction and possible subsequent autoimmune pathology. However, some of these mechanisms are also utilized by tumors to avoid immune recognition and attention of the immune cells. Other tumor escape mechanisms involve increased production of cytokines and factors which are responsible for immunosuppressive tumor microenvironment where effective immune response is actively blocked. This review summarizes the most frequently used strategies, which are utilized by tumors to avoid immune recognition and/or killing by the immune cells. Key words: immune evasion – tumor escape – immunotherapy – CTLA-4 – PD-1 – immune checkpoint * I declare that, in connection with this contribution of which I am the co-author, I have a conflict of interest with following company: Bristol-Myers Squibb al. s r. o. Author is former employee of Institute of Microbiology of the AS CR, v. v. i., Prague. ** The author declares she has no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 4. 8. 2015 Accepted: 1. 10. 2015

• Klíčová slova
• imunitní úniky, kontrolní body imunitní reakce, PD‑1, CTLA‑4, únikové mechanizmy nádorů, LAG-3, TIM-3,
• MeSH
• adaptivní imunita MeSH
• antigen CTLA-4 imunologie   MeSH
• antigeny CD279 imunologie   MeSH
• antigeny nádorové imunologie   MeSH
• CD antigeny imunologie   MeSH
• hostitel s imunodeficiencí MeSH
• imunitní dozor * imunologie   MeSH
• imunitní systém - jevy fyziologie   imunologie   MeSH
• imunoterapie MeSH
• lidé MeSH
• membránové proteiny imunologie   MeSH
• modely imunologické MeSH
• nádory * imunologie   MeSH
• přirozená imunita MeSH
• regulační T-lymfocyty imunologie   MeSH
• únik nádoru z imunitní kontroly * fyziologie   imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

BACKGROUND: The WNT/planar-cell-polarity (PCP) pathway is a key regulator of cell polarity and directional cell movements. Core PCP proteins such as Van Gogh-like2 (VANGL2) are evolutionarily highly conserved; however, the mammalian PCP machinery is still poorly understood mainly due to lack of suitable models and quantitative methodology. WNT/PCP has been implicated in many human diseases with the most distinguished positive role in the metastatic process, which accounts for more than 90% of cancer related deaths, and presents therefore an attractive target for pharmacological interventions. However, cellular assays for the assessment of PCP signaling, which would allow a more detailed mechanistic analysis of PCP function and possibly also high throughput screening for chemical compounds targeting mammalian PCP signaling, are still missing. RESULTS: Here we describe a mammalian cell culture model, which correlates B lymphocyte migration of patient-derived MEC1 cells and asymmetric localization of fluorescently-tagged VANGL2. We show by live cell imaging that PCP proteins are polarized in MEC1 cells and that VANGL2 polarization is controlled by the same mechanism as in tissues i.e. it is dependent on casein kinase 1 activity. In addition, destruction of the actin cytoskeleton leads to migratory arrest and cell rounding while VANGL2-EGFP remains polarized suggesting that active PCP signaling visualized by polarized distribution of VANGL2 is a cause for and not a consequence of the asymmetric shape of a migrating cell. CONCLUSIONS: The presented imaging-based methodology allows overcoming limitations of earlier approaches to study the mammalian WNT/PCP pathway, which required in vivo models and analysis of complex tissues. Our system investigating PCP-like signaling on a single-cell level thus opens new possibilities for screening of compounds, which control asymmetric distribution of proteins in the PCP pathway.

• MeSH
• B-lymfocyty metabolismus   patologie   MeSH
• chronická lymfatická leukemie genetika   imunologie   patologie   MeSH
• intracelulární signální peptidy a proteiny genetika   imunologie   MeSH
• lidé MeSH
• membránové proteiny genetika   imunologie   MeSH
• nádorové buněčné linie MeSH
• pohyb buněk genetika   imunologie   MeSH
• polarita buněk genetika   imunologie   MeSH
• signální dráha Wnt genetika   imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH