Background and aims: The majority of colorectal cancers arise from detectable adenomatous or serrated lesions. Here we demonstrate how deregulated alternative splicing of CD44 gene in diseased colon mucosa results in downregulation of standard isoform of CD44 gene (CD44s) and upregulation of variant isoform CD44v8-10. Our aim is to show that upregulation of CD44v8-10 isoform is a possible marker of precancerous lesion in human colon. Methods: We analysed pairs of fresh biopsy specimen of large intestine in a cohort of 50 patients. We studied and compared alternative splicing profile of CD44 gene in colon polyps and adjoined healthy colon mucosa. We performed end-point and qRT PCR, western blotting, IHC staining and flow cytometry analyses. Results: We detected more than five-fold overexpression of CD44v8-10 isoform and almost twenty-fold downregulation of standard isoform CD44s in colon polyps compared to adjoined healthy tissue with p = 0.018 and p < 0.001 in a cohort of 50 patients. Our results also show that aberrant splicing of CD44 occurs in both biologically distinct subtypes of colorectal adenoma possibly in ESRP-1 specific manner. Conclusion: 92% of the colon polyp positive patients overexpressed CD44v8-10 isoform in their colon polyps while only 36% of them had positive fecal occult blood test which is currently a standard non-invasive screening technique. Impact: We believe that our results are important for further steps leading to application of CD44v8-10 isoform as a biomarker of colorectal precancerosis in non-invasive detection. Early detection of colon precancerosis means successful prevention of colorectal carcinoma.

• MeSH
• antigeny CD44 genetika   metabolismus   MeSH
• kolon metabolismus   patologie   MeSH
• kolorektální nádory diagnóza   genetika   metabolismus   MeSH
• lidé MeSH
• nádorové biomarkery genetika   metabolismus   MeSH
• polypy tlustého střeva metabolismus   patologie   MeSH
• prognóza MeSH
• protein - isoformy MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• alergeny aplikace a dávkování   imunologie   MeSH
• aplikace slizniční MeSH
• imunologická tolerance MeSH
• ovalbumin aplikace a dávkování   imunologie   MeSH
• prasata MeSH
• sublinguální imunoterapie metody   MeSH
• systémy cílené aplikace léků MeSH
• ústní spodina MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH

Development of vaccine preventing HIV-1 infection is hindered by high variability of envelope glycoprotein (Env), known target for neutralizing antibodies. Identification of several monoclonal antibodies neutralizing broad range of HIV-1 variants (bn-mAbs) recognizing Env glycans as a part of their epitopes provides the new opportunity for vaccine design. As the composite glycans-containing epitopes are less effective immunogens than protein epitopes, we propose to develop a set of peptides mimicking epitopes recognized by above bn-mAbs using high-affinity binders approach developed by the co-applicant. Binders will be selected based on specific interaction with respective bn-mAbs, fused to albumin, and used for immunization of animals in form of proteoliposomes or DNA vaccines. Sera will be characterized concerning to their specificity, affinity, and neutralization activity. This approach provides the possibility to identify the candidate antigen for future HIV-1 vaccine and provide a platform for development of glycan-mimicking vaccines eliciting good immunological memory.

Vývoj vakcíny proti HIV-1 infekci je brzděn enormní variabilitou obalového glykoproteinu (Env), cíle neutralizačních protilátek. Identifikace sady monoklonálních protilátek neutralizujících široké spektrum variant HIV-1 (bn-mAb), reagujících s Env glykany, jako součástí jejich epitopu, nabízí novou cestu vývoje vakcíny. Jelikož kompozitní epitopy obsahující glykany jsou méně efektivní imunogeny než proteinové epitopy navrhujeme vyvinout sadu peptidů napodobujících epitopy rozlišované výše popsanými bn-mAb pomocí metody vysoce afinitních ligandů (binderů) vyvinutých spolunavrhovatelem projektu. Bindery budou selektovány na základě jejich specifické vazby na jednotlivé bn-mAb, budou fúzovány s albuminem a kotvami a ve formě proteoliposomů nebo DNA vakcín užity k imunizaci experimentálních zvířat. Poté bude charakterizována specificita, afinita a neutralizační aktivita sérových protilátek. Přístup skýtá možnost identifikovat kandidátní antigen pro vakcínu proti HIV-1 a představuje platformu pro vývoj vakcín napodobujících glykanové epitopy navozujících dobrou imunologickou paměť.

• MeSH
• epitopy terapeutické užití   MeSH
• experimenty na zvířatech MeSH
• HIV infekce terapie   MeSH
• HIV obalový protein gp120 MeSH
• HIV-1 MeSH
• imunizace MeSH
• látky proti HIV MeSH
• ligandy MeSH
• neutralizující protilátky MeSH
• rekombinantní proteiny terapeutické užití   MeSH
• vakcíny proti AIDS MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• dermatovenerologie
• infekční lékařství
• alergologie a imunologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

The natural adjuvant properties of bacterial ghosts (BGs) lie within the presence of intact pathogen-associated molecular patterns on their surface. BGs can improve the direct delivery, natural processing and presentation of target antigens within dendritic cells (DCs). Moreover, sensitization of human DCs by cancer cell lysate (oncolysate)-loaded BGs in the presence of IFN-α and GM-CSF enhanced DC maturation as indicated by an increased expression of maturation markers and co-stimulatory molecules, higher production of IL-12p70 and stimulation of significantly increased proliferation of both autologous CD4(+) and CD8(+) T cells compared to DCs matured in the presence of purified lipopolysaccharide. The induced T cells efficiently recognized oncolysate-derived tumor-associated antigens expressed by cancer cells used for the production of oncolysate. Our optimized one-step simultaneous antigen delivery and DC maturation-inducing method emerges as a promising tool for the development and implementation of next-generation cellular cancer immunotherapies.

• MeSH
• buněčná diferenciace imunologie   MeSH
• CD4-pozitivní T-lymfocyty imunologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• dendritické buňky imunologie   mikrobiologie   transplantace   MeSH
• Escherichia coli imunologie   MeSH
• fenotyp MeSH
• glioblastom imunologie   terapie   MeSH
• imunoterapie adoptivní metody   MeSH
• interleukin-12 biosyntéza   imunologie   MeSH
• lidé MeSH
• lipopolysacharidy farmakologie   MeSH
• nádorové buněčné linie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Nanofibre-based mucoadhesive films were invented for oromucosal administration of nanocarriers used for delivery of drugs and vaccines. The mucoadhesive film consists of an electrospun nanofibrous reservoir layer, a mucoadhesive film layer and a protective backing layer. The mucoadhesive layer is responsible for tight adhesion of the whole system to the oral mucosa after application. The electrospun nanofibrous reservoir layer is intended to act as a reservoir for polymeric and lipid-based nanoparticles, liposomes, virosomes, virus-like particles, dendrimers and the like, plus macromolecular drugs, antigens and/or allergens. The extremely large surface area of nanofibrous reservoir layers allows high levels of nanoparticle loading. Nanoparticles can either be reversibly adsorbed to the surface of nanofibres or they can be deposited in the pores between the nanofibres. After mucosal application, nanofibrous reservoir layers are intended to promote prolonged release of nanoparticles into the submucosal tissue. Reversible adsorption of model nanoparticles as well as sufficient mucoadhesive properties were demonstrated. This novel system appears appropriate for the use in oral mucosa, especially for sublingual and buccal tissues. To prove this concept, trans-/intramucosal and lymph-node delivery of PLGA-PEG nanoparticles was demonstrated in a porcine model. This system can mainly be used for sublingual immunization and the development of "printed vaccine technology".

• MeSH
• adheziva chemie   MeSH
• aplikace bukální MeSH
• aplikace sublinguální MeSH
• léčivé přípravky aplikace a dávkování   MeSH
• liposomy chemie   MeSH
• lymfatické uzliny metabolismus   MeSH
• myši MeSH
• nanočástice chemie   MeSH
• nanovlákna chemie   MeSH
• polyethylenglykoly chemie   MeSH
• polyglactin 910 chemie   MeSH
• prasata MeSH
• systémy cílené aplikace léků metody   MeSH
• ústní sliznice metabolismus   MeSH
• vakcinace metody   MeSH
• vakcíny aplikace a dávkování   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Pseudorabies virus (PrV), a causative agent of Aujeszky's disease, is deadly to most mammals with the exception of higher primates and men. This disease causes serious economic loses among farm animals, especially pigs, yet many European countries are today claimed to be Aujeszky's disease free because of the discovery of an efficient vaccination for pigs. In reality, the virus is still present in wild boar. Current vaccines are neither suitable for dogs nor are there anti-PrV drugs approved for veterinary use. Therefore, the disease still represents a high threat, particularly for expensive hunting dogs that can come into close contact with infected boars. Here we report on the anti-PrV activities of a series of synthetic diaminopurine-based acyclic nucleoside phosphonate (DAP-ANP) analogues. Initially, all synthetic DAP-ANPs under investigation are shown to exhibit minimal cytotoxicity by MTT and XTT tests (1-100μM range). Thereafter in vitro infection models are established using PrV virus SuHV-1, optimized on PK-15 and RK-13 cell lines. Out of the six DAP-ANP analogues tested, analogue VI functionalized with a cyclopropyl group on the 6-amino position of the purine ring proves the most effective antiviral DAP-ANP analogue against PrV infection, aided by sufficient hydrophobic character to enhance bioavailability to its cellular target viral DNA-polymerase. Four other DAP-ANP analogues with functional groups introduced to the C2'position are shown ineffective against PrV infection, even with favourable hydrophobic properties. Cidofovir(®), a drug approved against various herpesvirus infections, is found to exert only low activity against PrV in these same in vitro models.

• MeSH
• 2-aminopurin analogy a deriváty   chemie   farmakologie   MeSH
• antivirové látky chemie   farmakologie   MeSH
• buněčné linie MeSH
• buňky MDCK MeSH
• Herpesviridae účinky léků   MeSH
• organofosfonáty chemie   farmakologie   MeSH
• pseudovzteklina farmakoterapie   MeSH
• psi MeSH
• replikace DNA účinky léků   MeSH
• techniky in vitro MeSH
• transmisní elektronová mikroskopie MeSH
• Vero buňky MeSH
• zvířata MeSH
• Check Tag
• psi MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakt z konference MeSH

Hsp90-CA is present in cell wall of Candida pseudohyphae or hyphae-typical pathogenic morphotype for both systemic and mucosal Candida infections. Heat shock protein from Candida albicans (hsp90-CA) is an important target for protective antibodies during disseminated candidiasis of experimental mice and human. His-tagged protein rHsp90 was prepared and used as the antigen for preparation of experimental recombinant liposomal vaccine. Nickel-chelating liposomes (the size around 100nm, PDI≤0.1) were prepared from the mixture of egg phosphatidyl choline and nickel-chelating lipid DOGS-NTA-Ni (molar ratio 95:5%) by hydration of lipid film and extrusion methods. New non-pyrogenic hydrophobised derivative of MDP (C18-O-6-norAbuMDP) was incorporated into liposomes as adjuvans. rHsp90 was attached onto the surface of metallochelating liposomes by metallochelating bond and the structure of these proteoliposomes was studied by dynamic light scattering, AF microscopy, TEM and GPC. The liposomes with surface-exposed C18-O-6-norAbuMDP were well recognised and phagocyted by human dendritic cells in vitro. In vivo the immune response towards this experimental vaccine applied in mice (i.d.) demonstrated both TH1 and TH2 response comparable to FCA, but without any side effects. Metallochelating liposomes with lipophilic derivatives of muramyl dipeptide represent a new biocompatible platform for construction of experimental recombinant vaccines and drug-targeting systems.

• MeSH
• antigeny fungální imunologie   metabolismus   MeSH
• biokompatibilní potahované materiály metabolismus   MeSH
• buněčná imunita MeSH
• Candida imunologie   MeSH
• chelátory chemie   metabolismus   MeSH
• dendritické buňky imunologie   metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• liposomy MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nikl imunologie   metabolismus   MeSH
• proteiny tepelného šoku HSP90 imunologie   metabolismus   MeSH
• syntetické vakcíny imunologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Paclitaxel (PTX) is approved for the treatment of ovarian and breast cancer. The commercially available preparation of PTX, Cremophor EL(R) is associated with hypersensitivity reactions in spite of a suitable premedication. In general, the developed liposomal PTX formulations are troubled with low PTX encapsulation capacity (maximal content, 3 mol%) and accompanied by PTX crystallisation. The application of "pocket-forming" lipids significantly increased the encapsulation capacity of PTX in the liposomes up to 10 mol%. Stable lyophilised preparation of PTX (7 mol%) encapsulated in the liposomes composed of SOPC/POPG/MOPC (molar ratio, 60:20:20) doped with 5 mol% vitamin E had the size distribution of 180-190 nm (PDI, 0.1) with zeta-potential of -31 mV. Sucrose was found to be a suitable cryoprotectant at the lipid:sugar molar ratios of 1:5-1:10. This liposomal formulation did not show any evidence of toxicity in C57BL/6 mice treated with the highest doses of PTX (100 mg/kg administered as a single dose and 150 mg/kg as a cumulative dose applied in three equivalent doses in 48-h intervals). A dose-dependent anticancer effect was found in both hollow fibre implants and syngenic B16F10 melanoma mouse tumour models.

• MeSH
• antitumorózní látky fytogenní aplikace a dávkování   chemie   terapeutické užití   toxicita   MeSH
• lipidy chemie   MeSH
• liposomy MeSH
• lyofilizace MeSH
• melanom experimentální farmakoterapie   metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nanotechnologie MeSH
• paclitaxel aplikace a dávkování   chemie   terapeutické užití   toxicita   MeSH
• příprava léků MeSH
• stabilita léku MeSH
• velikost částic MeSH
• viabilita buněk účinky léků   MeSH
• xenogenní modely - testy antitumorózní aktivity MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH