The Turkevich method was optimized to prepare gold nanoparticles (AuNP) stabilized by polyethyleneglycol (PEG) for μCT. Using various independent modalities, we thoroughly characterized the optimized PEG-AuNPs. Here, we show that PEG-AuNPs are retained in the blood and provide a high contrast in the high-resolution μCT imaging of blood vessels and inner organs. The biodistribution is characterized by prolonged circulation in the blood and accumulation in the liver, spleen and skin. The accumulation of AuNP in the skin resulted in the blue discoloration of eyes and the whole skin. In vitro experiments using a leukemic monocyte THP-1 cell line model expressing high levels of NLRP3 demonstrated that the NLRP3inflammasome was not activated by PEG AuNP. Over 9 months, the mice were scanned by μCT and were in good health. Scans in mice using PEG-stabilized AuNPs in this study were sharper, with a higher contrast, when compared to a commercial contrasting agent at the same dose. The PEG-AuNPs were morphologically and chemically stable for at least two years when stored in the refrigerator.

• Publikační typ
• časopisecké články MeSH

SARS-CoV-2 je virus, který infikuje respirační trakt a může způsobit závažné, někdy až život ohrožující onemocnění COVID-19. U více než 5 % symptomatických pacientů je infekce spojena s výskytem postakutních následků. První kontakt viru s imunitním systémem nazofaryngu a orofaryngu vyvolává slizniční imunitní odpověď spojenou s lokální produkcí sekrečních protilátek třídy imunoglobulinů A (sIgA), což může přispět k lokalizaci viru na horní cesty dýchací a asymptomatické či pouze klinicky mírné infekci. Během pandemie rychle vyvinuté systémově působící vakcíny úspěšně chrání před závažnou formou onemocnění a jeho postakutními následky, nicméně neindukují protilátky ve slizničních sekretech u dosud SARS-CoV-2-naivních jedinců. U těch, kteří již infekci prodělali, mohou přesto systémové vakcíny posílit tvorbu protilátek sIgA. Je třeba zdůraznit, že klinická prospěšnost systémové imunizace vakcínami proti COVID-19 přesvědčivě dokumentovaná u desítek milionů jedinců zastiňuje vzácné, mnohdy sporně dokumentované komplikace očkování. Neschopnost současných vakcín navodit slizniční imunitní odpověď a zabránit šíření viru sekrety infikovaných jedinců poukazuje na vzájemnou nezávislost slizniční a systémové imunity. To podtrhuje potřebu vývoje vakcín, které vyvolají patřičné odpovědi v obou kompartmentech imunitního systému.

SARS-CoV-2 is a virus which infects the respiratory tract and may cause severe, occasionally life-threatening disease COVID-19. In more than 5% of symptomatic patients the infection is associated with post-acute symptoms. The initial contact of the virus with the immune system of the nasopharynx and oropharynx induces a mucosal immune response manifested by the production of secretory IgA (sIgA) antibodies which may contribute to the restriction of the infection to the upper respiratory tract and an asymptomatic or clinically mild disease. The current systemically administered vaccines protected against the severe COVID-19 infection and its post-acute sequelae. However, they do not induce antibodies in mucosal secretions in SARS-CoV-2-naive individuals. In contrast, in those who previously experienced mucosal infection, systemically administered vaccines may stimulate sIgA production. The clinical benefit of systemic vaccination convincingly documented in tens of millions of individuals overshadows the rare, sometimes controversial reports of complications encountered after vaccination. The inability of current SARS-CoV-2 vaccines to induce mucosal immune responses and to prevent the spreading of the virus by external secretions demonstrates the mutual independence of mucosal and systemic compartments of the immune system, and thus emphasizes need for the development of vaccines inducing protective immune responses in both compartments.

• MeSH
• COVID-19 imunologie   prevence a kontrola   MeSH
• lidé MeSH
• slizniční imunita imunologie   účinky léků   MeSH
• vakcinace MeSH
• vakcíny proti COVID-19 * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Lyme disease is an infection caused by spirochetes of the species complex Borrelia burgdorferi. It is multisystem disease most commmonly manifested as neurological or rheumatic symptoms. In some patients chronic complications persist after treatment. This condition is ussually referred as Post-Lyme disease syndrome and its pathogenesis is unclear. Both the persistence of bacteria and the involvement of autoimmune mechanisms are discussed as a possible cause. A combination of both factors in the form of chronic stimulation of the immune system by persistent inactive forms of Borrelia is also contemplated. The persistence of bacteria in the organism can be facilitated by the non-spiral forms of Borrelia, which has been observed when Borrelia culture is exposed to unfavorable conditions. The project will be focused on the mechanisms of formation of non-spiral forms of Borrelia spirochetes. We will test their infectivity and immunogenicity in vivo and interactions with mammalian cells in vitro. Results obtained during the project has the potential to contribute to a more effective diagnosis and treatment of Lyme borreliosis.

Lymeská borelioza je bakteriální multisystémové onemocnění způsobené spirochetami druhového komplexu Borrelia burgdorferi. Projevuje se nejčastěji kožními, neurologickými nebo kloubními symptomy. Neléčené onemocnění může přecházet do pozdních stádií s chronickým postižením. Přestože u většiny nemocných je efektivní antibiotická léčba, u části pacientů se i přes intenzivní antibakteriální terapii objevují přetrvávající potíže. Tento stav bývá uváděn jako tzv. post-boreliový syndrom. Příčina tohoto stavu dosud není vyjasněna, zvažuje se jak perzistence samotných bakterií, tak zapojení autoimunitních mechanismů. Uvažuje se i o kombinaci obou faktorů ve formě chronické stimulace imunitního systému perzistujícími neaktivními formami Borelií. Na perzistenci bakterií v organismu se mohou podílet nespirální formy Borelií, které byly pozorovány při vystavení Boreliové kultury nepřiznivým podmínkám. V projektu se zaměříme na mechanismy vzniku nespirálních forem Borelií. Budeme testovat jejich infekčnost a imunogenitu in vivo a interakce se savčími buňkami in vitro. Získané poznatky mají potenciál přispěk ke zefektivnění diagnostiky a léčby Lymeské boreliozy.

• Klíčová slova
• lyme disease, Borrelia burgdorferi, Borrelia burgdorferi, Lymská borelioza, Post-Treatment Lyme Disease Syndrome, Post-Treatment Lyme Disease Syndrome, nespirální formy Borrelií, non-spiral forms of Borrelia,

• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Human immunodeficiency virus type 1 (HIV-1) vaccine immunogens capable of inducing broadly neutralizing antibodies (bNAbs) remain obscure. HIV-1 evades immune responses through enormous diversity and hides its conserved vulnerable epitopes on the envelope glycoprotein (Env) by displaying an extensive immunodominant glycan shield. In elite HIV-1 viremic controllers, glycan-dependent bNAbs targeting conserved Env epitopes have been isolated and are utilized as vaccine design templates. However, immunological tolerance mechanisms limit the development of these antibodies in the general population. The well characterized bNAbs monoclonal variants frequently exhibit extensive levels of somatic hypermutation, a long third heavy chain complementary determining region, or a short third light chain complementarity determining region, and some exhibit poly-reactivity to autoantigens. This review elaborates on the obstacles to engaging and manipulating the Env glycoprotein as an effective immunogen and describes an alternative reverse vaccinology approach to develop a novel category of bNAb-epitope-derived non-cognate immunogens for HIV-1 vaccine design.

• MeSH
• epitopy imunologie   MeSH
• genové produkty env - virus lidské imunodeficience imunologie   MeSH
• HIV infekce imunologie   MeSH
• HIV protilátky * imunologie   MeSH
• HIV-1 * imunologie   MeSH
• lidé MeSH
• ligandy MeSH
• molekulární mimikry imunologie   MeSH
• neutralizující protilátky * imunologie   MeSH
• polysacharidy imunologie   MeSH
• vakcíny proti AIDS * imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Liposomes are one of the most important drug delivery vectors, nowadays used in clinics. In general, polyethylene glycol (PEG) is used to ensure the stealth properties of the liposomes. Here, we have employed hydrophilic, biocompatible and highly non-fouling N-(2-hydroxypropyl) methacrylamide (HPMA)-based copolymers containing hydrophobic cholesterol anchors for the surface modification of liposomes, which were prepared by the method of lipid film hydration and extrusion through 100 nm polycarbonate filters. Efficient surface modification of liposomes was confirmed by transmission electron microscopy, atomic force microscopy, and gradient ultracentrifugation. The ability of long-term circulation in the vascular bed was demonstrated in rabbits after i.v. application of fluorescently labelled liposomes. Compared to PEGylated liposomes, HPMA-based copolymer-modified liposomes did not induce specific antibody formation and did not activate murine and human complement. Compared with PEGylated liposomes, HPMA-based copolymer-modified liposomes showed a better long-circulating effect after repeated administration. HPMA-based copolymer-modified liposomes thus represent suitable new candidates for a generation of safer and improved liposomal drug delivery platforms.

• MeSH
• akrylamidy chemie   MeSH
• aktivace komplementu účinky léků   MeSH
• cholesterol chemie   krev   MeSH
• hydrofobní a hydrofilní interakce * MeSH
• králíci MeSH
• lidé MeSH
• liposomy * MeSH
• myši MeSH
• polyethylenglykoly * chemie   MeSH
• polymery chemie   MeSH
• povrchové vlastnosti * MeSH
• systémy cílené aplikace léků MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Programmed cell death 1 (PD-1) belongs to immune checkpoint proteins ensuring negative regulation of the immune response. In non-small cell lung cancer (NSCLC), the sensitivity to treatment with anti-PD-1 therapeutics, and its efficacy, mostly correlated with the increase of tumor infiltrating PD-1+ lymphocytes. Due to solid tumor heterogeneity of PD-1+ populations, novel low molecular weight anti-PD-1 high-affinity diagnostic probes can increase the reliability of expression profiling of PD-1+ tumor infiltrating lymphocytes (TILs) in tumor tissue biopsies and in vivo mapping efficiency using immune-PET imaging. METHODS: We designed a 13 kDa β-sheet Myomedin scaffold combinatorial library by randomization of 12 mutable residues, and in combination with ribosome display, we identified anti-PD-1 Myomedin variants (MBA ligands) that specifically bound to human and murine PD-1-transfected HEK293T cells and human SUP-T1 cells spontaneously overexpressing cell surface PD-1. RESULTS: Binding affinity to cell-surface expressed human and murine PD-1 on transfected HEK293T cells was measured by fluorescence with LigandTracer and resulted in the selection of most promising variants MBA066 (hPD-1 KD = 6.9 nM; mPD-1 KD = 40.5 nM), MBA197 (hPD-1 KD = 29.7 nM; mPD-1 KD = 21.4 nM) and MBA414 (hPD-1 KD = 8.6 nM; mPD-1 KD = 2.4 nM). The potential of MBA proteins for imaging of PD-1+ populations in vivo was demonstrated using deferoxamine-conjugated MBA labeled with 68Galium isotope. Radiochemical purity of 68Ga-MBA proteins reached values 94.7-99.3% and in vitro stability in human serum after 120 min was in the range 94.6-98.2%. The distribution of 68Ga-MBA proteins in mice was monitored using whole-body positron emission tomography combined with computerized tomography (PET/CT) imaging up to 90 min post-injection and post mortem examined in 12 mouse organs. The specificity of MBA proteins was proven by co-staining frozen sections of human tonsils and NSCLC tissue biopsies with anti-PD-1 antibody, and demonstrated their potential for mapping PD-1+ populations in solid tumors. CONCLUSIONS: Using directed evolution, we developed a unique set of small binding proteins that can improve PD-1 diagnostics in vitro as well as in vivo using PET/CT imaging.

• MeSH
• antigeny CD279 * metabolismus   MeSH
• HEK293 buňky MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory plic diagnostické zobrazování   patologie   metabolismus   genetika   MeSH
• nemalobuněčný karcinom plic diagnostické zobrazování   patologie   metabolismus   MeSH
• pozitronová emisní tomografie * metody   MeSH
• proteinové inženýrství * MeSH
• sekvence aminokyselin MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The initial phase of multiple sclerosis (MS), often known as clinically isolated syndrome (CIS), is a critical period for identifying individuals at high risk of progressing to full-blown MS and initiating timely treatment. In this study, we aimed to evaluate the prognostic value of C-X-C motif chemokine ligand 13 (CXCL13) and interleukin-8 (IL-8) as potential markers for CIS patients' conversion to MS. Our study encompassed patients with CIS, those with relapsing-remitting MS (RRMS), and control subjects, with sample analysis conducted on both cerebrospinal fluid (CSF) and serum. Patients were categorized into four groups: CIS-CIS (no MS development within 2 years), CIS-RRMS (conversion to RRMS within 2 years), RRMS (already diagnosed), and a control group (CG) with noninflammatory central nervous system disorders. Results showed significantly elevated levels of CXCL13 in CSF across all patient groups compared with the CG (p < 0.0001, Kruskal-Wallis test). Although CXCL13 concentrations were slightly higher in the CIS-RRMS group, statistical significance was not reached. Similarly, significantly higher levels of IL-8 were detected in CSF samples from all patient groups compared with the CG (p < 0.0001, Kruskal-Wallis test). Receiver operating characteristic analysis in the CIS-RRMS group identified both CXCL13 (area under receiver operating characteristic curve = .959) and IL-8 (area under receiver operating characteristic curve = .939) in CSF as significant predictors of CIS to RRMS conversion. In conclusion, our study suggests a trend towards elevated CSF IL-8 and CSF CXCL13 levels in CIS patients progressing to RRMS. These findings emphasize the importance of identifying prognostic markers to guide appropriate treatment strategies for individuals in the early stages of MS.

• MeSH
• biologické markery mozkomíšní mok   krev   MeSH
• chemokin CXCL13 * mozkomíšní mok   krev   MeSH
• demyelinizační nemoci mozkomíšní mok   MeSH
• dospělí MeSH
• interleukin-8 * mozkomíšní mok   krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• prognóza MeSH
• progrese nemoci * MeSH
• relabující-remitující roztroušená skleróza * mozkomíšní mok   krev   diagnóza   MeSH
• roztroušená skleróza mozkomíšní mok   krev   diagnóza   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

Borrelia burgdorferi sensu lato is a species complex of pleomorphic spirochetes, including species that cause Lyme disease (LD) in humans. In addition to classic spiral forms, these bacteria are capable of creating morphological forms referred to as round bodies and aggregates. The subject of discussion is their possible contribution to the persistence of infection or post-infection symptoms in LD. This study investigates the immunological properties of these forms by monitoring reactivity with early (n = 30) and late stage (n = 30) LD patient sera and evaluating the immune response induced by vaccination of mice. In patient sera, we found a quantitative difference in reactivity with individual morphotypes, when aggregates were recognized most intensively, but the difference was statistically significant in only half of the tested strains. In post-vaccination mouse sera, we observed a statistically significant higher reactivity with antigens p83 and p25 (OspC) in mice vaccinated with aggregates compared to mice vaccinated with spiral forms. The importance of the particulate nature of the antigen for the induction of a Th1-directed response has also been demonstrated. In any of morphological forms, the possibility of inducing antibodies cross-reacting with human nuclear and myositis specific/associated autoantigens was not confirmed by vaccination of mice.

• MeSH
• antigeny bakteriální MeSH
• Borrelia burgdorferi komplex * MeSH
• Borrelia burgdorferi * MeSH
• lidé MeSH
• lymeská nemoc * mikrobiologie   MeSH
• myši MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Non-small cell lung cancer (NSCLC) is largely promoted by a multistep tumorigenesis process involving various genetic and epigenetic alterations, which essentially contribute to the high incidence of mortality among patients with NSCLC. Clinical observations revealed that NSCLC also co-opts a multifaceted immune checkpoint dysregulation as an important driving factor in NSCLC progression and development. For example, a deregulated PI3K/AKT/mTOR pathway has been noticed in 50-70% of NSCLC cases, primarily modulated by mutations in key oncogenes such as ALK, EGFR, KRAS, and others. Additionally, genetic association studies containing patient-specific factors and local reimbursement criteria expose/reveal mutations in EGFR/ALK/ROS/BRAF/KRAS/PD-L1 proteins to determine the suitability of available immunotherapy or tyrosine kinase inhibitor therapy. Thus, the expression of such checkpoints on tumors and immune cells is pivotal in understanding the therapeutic efficacy and has been extensively studied for NSCLC treatments. Therefore, this review summarizes current knowledge in NSCLC tumorigenesis, focusing on its genetic and epigenetic intricacies, immune checkpoint dysregulation, and the evolving landscape of targeted therapies. In the context of current and future therapies, we emphasize the significance of antibodies targeting PD-1/PD-L1 and CTLA-4 interactions as the primary therapeutic strategy for immune system reactivation in NSCLC. Other approaches involving the promising potential of nanobodies, probodies, affibodies, and DARPINs targeting immune checkpoints are also described; these are under active research or clinical trials to mediate immune regulation and reduce cancer progression. This comprehensive review underscores the multifaceted nature, current state and future directions of NSCLC research and treatment.

• MeSH
• antigeny CD274 metabolismus   MeSH
• erbB receptory metabolismus   MeSH
• fosfatidylinositol-3-kinasy metabolismus   MeSH
• karcinogeneze MeSH
• lidé MeSH
• nádorová transformace buněk MeSH
• nádory plic * farmakoterapie   genetika   MeSH
• nemalobuněčný karcinom plic * farmakoterapie   genetika   MeSH
• protoonkogenní proteiny p21(ras) MeSH
• tyrosinkinasové receptory metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Lyme disease (LD) spirochetes are well known to be able to disseminate into the tissues of infected hosts, including humans. The diverse strategies used by spirochetes to avoid the host immune system and persist in the host include active immune suppression, induction of immune tolerance, phase and antigenic variation, intracellular seclusion, changing of morphological and physiological state in varying environments, formation of biofilms and persistent forms, and, importantly, incursion into immune-privileged sites such as the brain. Invasion of immune-privileged sites allows the spirochetes to not only escape from the host immune system but can also reduce the efficacy of antibiotic therapy. Here we present a case of the detection of spirochetal DNA in multiple loci in a LD patient's post-mortem brain. The presence of co-infection with Borrelia burgdorferi sensu stricto and Borrelia garinii in this LD patient's brain was confirmed by PCR. Even though both spirochete species were simultaneously present in human brain tissue, the brain regions where the two species were detected were different and non-overlapping. The presence of atypical spirochete morphology was noted by immunohistochemistry of the brain samples. Atypical morphology was also found in the tissues of experimentally infected mice, which were used as a control.

• MeSH
• Borrelia burgdorferi komplex * genetika   MeSH
• Borrelia burgdorferi * genetika   MeSH
• Borrelia * genetika   MeSH
• lidé MeSH
• lymeská nemoc * MeSH
• mozek MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH