Tumor-specific neoantigens can be highly immunogenic, but their identification for each patient and the production of personalized cancer vaccines can be time-consuming and prohibitively expensive. In contrast, tumor-associated antigens are widely expressed and suitable as an off the shelf immunotherapy. Here, we developed a PLGA-based nanoparticle vaccine that contains both the immunogenic cancer germline antigen NY-ESO-1 and an α-GalCer analog IMM60, as a novel iNKT cell agonist and dendritic cell transactivator. Three peptide sequences (85-111, 117-143, and 157-165) derived from immunodominant regions of NY-ESO-1 were selected. These peptides have a wide HLA coverage and were efficiently processed and presented by dendritic cells via various HLA subtypes. Co-delivery of IMM60 enhanced CD4 and CD8 T cell responses and antibody levels against NY-ESO-1 in vivo. Moreover, the nanoparticles have negligible systemic toxicity in high doses, and they could be produced according to GMP guidelines. Together, we demonstrated the feasibility of producing a PLGA-based nanovaccine containing immunogenic peptides and an iNKT cell agonist, that is activating DCs to induce antigen-specific T cell responses.

• MeSH
• B-lymfocyty imunologie   MeSH
• CD4-pozitivní T-lymfocyty imunologie   MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• kopolymer kyseliny glykolové a mléčné chemie   farmakologie   MeSH
• lidé MeSH
• nádorové proteiny chemie   farmakologie   MeSH
• nanočástice chemie   terapeutické užití   MeSH
• nosiče léků chemie   farmakologie   MeSH
• peptidové fragmenty chemie   farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Obesity, diabetes, insulin resistance, sedentary lifestyle, and Western diet are the key factors underlying non-alcoholic fatty liver disease (NAFLD), one of the most common liver diseases in developed countries. In many cases, NAFLD further progresses to non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and to hepatocellular carcinoma. The hepatic lipotoxicity and non-liver factors, such as adipose tissue inflammation and gastrointestinal imbalances were linked to evolution of NAFLD. Nowadays, the degree of adipose tissue inflammation was shown to directly correlate with the severity of NAFLD. Consumption of higher caloric intake is increasingly emerging as a fuel of metabolic inflammation not only in obesity-related disorders but also NAFLD. However, multiple causes of NAFLD are the reason why the mechanisms of NAFLD progression to NASH are still not well understood. In this review, we explore the role of food intake regulating peptides in NAFLD and NASH mouse models. Leptin, an anorexigenic peptide, is involved in hepatic metabolism, and has an effect on NAFLD experimental models. Glucagon-like peptide-1 (GLP-1), another anorexigenic peptide, and GLP-1 receptor agonists (GLP-1R), represent potential therapeutic agents to prevent NAFLD progression to NASH. On the other hand, the deletion of ghrelin, an orexigenic peptide, prevents age-associated hepatic steatosis in mice. Because of the increasing incidence of NAFLD and NASH worldwide, the selection of appropriate animal models is important to clarify aspects of pathogenesis and progression in this field.

• MeSH
• hypoglykemika farmakologie   MeSH
• lidé MeSH
• modely nemocí na zvířatech * MeSH
• nealkoholová steatóza jater farmakoterapie   etiologie   patofyziologie   MeSH
• peptidové fragmenty farmakologie   MeSH
• přijímání potravy * MeSH
• progrese nemoci MeSH
• regulace chuti k jídlu účinky léků   MeSH
• zánět komplikace   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Galanin and galanin receptors (GalRs) have been reported to be involved in the transmission and modulation of nociceptive information in the central nervous system (CNS). However, the underlying mechanism of the antinociception of GalRs in neuropathic pain remains unclear. This study investigated the antinociception induced by galanin receptor 1 (GalR1) via protein kinase A (PKA) signaling pathway in the nucleus accumbens (NAc) of rats with neuropathic pain. A mononeuropathy model was replicated by ligation of the left sciatic nerve, following which the expression of phospho-PKA (p-PKA) in the NAc were markedly up-regulated at 14(th) and 28(th) day after ligation of sciatic nerve, and p-PKA expression was down-regulated by intra-NAc injection of GalR1 agonist M617, but the GalR1 antagonist M35 did not have an effect. We also found that M35 in the NAc blocked the M617-induced increase in the hind paw withdrawal latencies (HWLs) of rats with mononeuropathy, but M35 alone had no effect on HWLs, and PKA inhibitor H-89 attenuated the M617-induced an increase in the HWLs. These results suggested that GalR1 induced an antinociception via inhibiting PKA activation, implying that GalR agonists may be potential and potent therapeutic options to treat chronic neuropathic pain.

• MeSH
• aktivace enzymů účinky léků   fyziologie   MeSH
• analgetika metabolismus   MeSH
• bradykinin analogy a deriváty   farmakologie   MeSH
• galanin analogy a deriváty   farmakologie   MeSH
• krysa rodu rattus MeSH
• měření bolesti účinky léků   metody   MeSH
• neuralgie metabolismus   prevence a kontrola   MeSH
• nucleus accumbens účinky léků   metabolismus   MeSH
• peptidové fragmenty farmakologie   MeSH
• potkani Sprague-Dawley MeSH
• proteinkinasy závislé na cyklickém AMP antagonisté a inhibitory   metabolismus   MeSH
• receptor galaninu typ 1 agonisté   antagonisté a inhibitory   biosyntéza   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Endothelin B (ET(B)) receptors present in abundance the central nervous system (CNS) have been shown to have significant implications in its development and neurogenesis. We have targeted ET(B) receptors stimulation using a highly specific agonist, IRL-1620, to treat CNS disorders. In a rat model of cerebral ischemia intravenous administration IRL-1620 significantly reduced infarct volume and improved neurological and motor functions compared to control. This improvement, in part, is due to an increase in neuroregeneration. We also investigated the role of IRL-1620 in animal models of Alzheimer's disease (AD). IRL-1620 improved learning and memory, reduced oxidative stress and increased VEGF and NGF in Abeta treated rats. IRL-1620 also improved learning and memory in an aged APP/PS1 transgenic mouse model of AD. These promising findings prompted us to initiate human studies. Successful chemistry, manufacturing and control along with mice, rat and dog toxicological studies led to completion of a human Phase I study in healthy volunteers. We found that a dose of 0.6 microg/kg of IRL-1620 can be safely administered, three times every four hours, without any adverse effect. A Phase II clinical study with IRL-1620 has been initiated in patients with cerebral ischemia and mild to moderate AD.

• MeSH
• endoteliny farmakologie   terapeutické užití   MeSH
• klinické zkoušky jako téma metody   MeSH
• lidé MeSH
• nemoci centrálního nervového systému farmakoterapie   metabolismus   MeSH
• neurodegenerativní nemoci farmakoterapie   metabolismus   MeSH
• oxidační stres účinky léků   fyziologie   MeSH
• peptidové fragmenty farmakologie   terapeutické užití   MeSH
• preklinické hodnocení léčiv metody   MeSH
• receptor endotelinu B agonisté   metabolismus   MeSH
• regenerace nervu účinky léků   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Cachexia represents a fatal energy-wasting syndrome in a large number of patients with cancer that mostly results in a pathological loss of skeletal muscle and adipose tissue. Here we show that tumor cell exposure and tumor growth in mice triggered a futile energy-wasting cycle in cultured white adipocytes and white adipose tissue (WAT), respectively. Although uncoupling protein 1 (Ucp1)-dependent thermogenesis was dispensable for tumor-induced body wasting, WAT from cachectic mice and tumor-cell-supernatant-treated adipocytes were consistently characterized by the simultaneous induction of both lipolytic and lipogenic pathways. Paradoxically, this was accompanied by an inactivated AMP-activated protein kinase (Ampk), which is normally activated in peripheral tissues during states of low cellular energy. Ampk inactivation correlated with its degradation and with upregulation of the Ampk-interacting protein Cidea. Therefore, we developed an Ampk-stabilizing peptide, ACIP, which was able to ameliorate WAT wasting in vitro and in vivo by shielding the Cidea-targeted interaction surface on Ampk. Thus, our data establish the Ucp1-independent remodeling of adipocyte lipid homeostasis as a key event in tumor-induced WAT wasting, and we propose the ACIP-dependent preservation of Ampk integrity in the WAT as a concept in future therapies for cachexia.

• MeSH
• bílá tuková tkáň účinky léků   metabolismus   MeSH
• bílé tukové buňky účinky léků   metabolismus   MeSH
• kachexie etiologie   metabolismus   MeSH
• kultivované buňky MeSH
• lipogeneze účinky léků   MeSH
• lipolýza účinky léků   MeSH
• metabolismus lipidů účinky léků   MeSH
• myši MeSH
• nádory komplikace   metabolismus   MeSH
• peptidové fragmenty farmakologie   MeSH
• proteinkinasy aktivované AMP metabolismus   farmakologie   MeSH
• proteiny regulující apoptózu účinky léků   metabolismus   MeSH
• techniky in vitro MeSH
• termogeneze účinky léků   MeSH
• uncoupling protein 1 účinky léků   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Previous studies from Multhoff and colleagues reported that plasma membrane Hsp70 acts as a tumour-specific recognition structure for activated NK cells, and that the incubation of NK cells with Hsp70 and/or a 14-mer peptide derived from the N-terminal sequence of Hsp70 (TKDNNLLGRFELSG, TKD, aa 450-463) plus a low dose of IL-2 triggers NK cell proliferation and migration, and their capacity to kill cancer cells expressing membrane Hsp70. Herein, we have used flow cytometry to determine the influence of in vitro stimulation of peripheral blood mononuclear cells from healthy individuals with IL-2 or IL-15, either alone or in combination with TKD peptide on the cell surface expression of CD94, NK cell activatory receptors (CD16, NK2D, NKG2C, NKp30, NKp44, NKp46, NKp80, KIR2DL4, DNAM-1 and LAMP1) and NK cell inhibitory receptors (NKG2A, KIR2DL2/L3, LIR1/ILT-2 and NKR-P1A) by CD3+CD56+ (NKT), CD3+CD4+, CD3+CD8+ and CD19+ populations. NKG2D, DNAM-1, LAMP1 and NKR-P1A expression was upregulated after the stimulation with IL-2 or IL-15 alone or in combination with TKD in NKT, CD8+ T cells and B cells. CD94 was upregulated in NKT and CD8+ T cells. Concurrently, an increase in a number of CD8+ T cells expressing LIR1/ILT-2 and CD4+ T cells positive for NKR-P1A was observed. The proportion of CD8+ T cells that expressed NKG2D was higher after IL-2/TKD treatment, when compared with IL-2 treatment alone. In comparison with IL-15 alone, IL-15/TKD treatment increased the proportion of NKT cells that were positive for CD94, LAMP1 and NKRP-1A. The more potent effect of IL-15/TKD on cell surface expression of NKG2D, LIR1/ILT-2 and NKRP-1A was observed in B cells compared with IL-15 alone. However, this increase was not of statistical significance. IL-2/TKD induced significant upregulation of LAMP1 in CD8+ T cells compared with IL-2 alone. Besides NK cells, other immunocompetent cells present within the fraction of peripheral blood mononuclear cells were influenced by the treatment with low-dose interleukins themselves or in combination with hsp70 derived (TKD) peptide.

• MeSH
• B-lymfocyty účinky léků   metabolismus   MeSH
• buňky NK účinky léků   metabolismus   MeSH
• dospělí MeSH
• interleukin-15 farmakologie   MeSH
• interleukin-2 farmakologie   MeSH
• kohortové studie MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• peptidové fragmenty chemie   farmakologie   MeSH
• proteiny tepelného šoku HSP70 chemie   metabolismus   MeSH
• receptory imunologické metabolismus   MeSH
• T-lymfocyty účinky léků   metabolismus   MeSH
• techniky in vitro MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Engineered combinatorial libraries derived from small protein scaffolds represent a powerful tool for generating novel binders with high affinity, required specificity and designed inhibitory function. This work was aimed to generate a collection of recombinant binders of human interleukin-23 receptor (IL-23R), which is a key element of proinflammatory IL-23-mediated signaling. A library of variants derived from the three-helix bundle scaffold of the albumin-binding domain (ABD) of streptococcal protein G and ribosome display were used to select for high-affinity binders of recombinant extracellular IL-23R. A collection of 34 IL-23R-binding proteins (called REX binders), corresponding to 18 different sequence variants, was used to identify a group of ligands that inhibited binding of the recombinant p19 subunit of IL-23, or the biologically active human IL-23 cytokine, to the recombinant IL-23R or soluble IL-23R-IgG chimera. The strongest competitors for IL-23R binding in ELISA were confirmed to recognize human IL-23R-IgG in surface plasmon resonance experiments, estimating the binding affinity in the sub- to nanomolar range. We further demonstrated that several REX variants bind to human leukemic cell lines K-562, THP-1 and Jurkat, and this binding correlated with IL-23R cell-surface expression. The REX125, REX009 and REX128 variants competed with the p19 protein for binding to THP-1 cells. Moreover, the presence of REX125, REX009 and REX115 variants significantly inhibited the IL-23-driven expansion of IL-17-producing primary human CD4(+) T-cells. Thus, we conclude that unique IL-23R antagonists derived from the ABD scaffold were generated that might be useful in designing novel anti-inflammatory biologicals.

• MeSH
• antiflogistika chemie   farmakologie   MeSH
• bakteriální proteiny chemie   MeSH
• buňky K562 MeSH
• buňky Th17 účinky léků   metabolismus   MeSH
• imunologické faktory chemie   farmakologie   MeSH
• interleukin-23 chemie   fyziologie   MeSH
• Jurkat buňky MeSH
• kompetitivní vazba MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• peptidové fragmenty chemie   farmakologie   MeSH
• preklinické hodnocení léčiv MeSH
• receptory interleukinů antagonisté a inhibitory   fyziologie   MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie aminokyselin MeSH
• vazba proteinů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In DNA vaccination, CD4(+) T-cell help can be enhanced by fusion of a gene encoding an immunization protein with a foreign gene or its part providing T(h) epitopes. To study the effect of helper epitope localization in a protein molecule, the influence of the vicinity of the helper epitope, and the impact of chimeric protein cellular localization, we fused the helper epitope p30 from tetanus toxin (TT, aa 947-967) with the N- or C-terminus of the mutated E7 oncoprotein (E7GGG) of human papillomavirus type 16, enlarged the p30 epitope with the flanking residues containing potential protease-sensitive sites and altered the cellular localization of the fusion constructs by signal sequences. The p30 epitope enhanced the E7-specific response, but only in constructs without added signal sequences. After localization of the fusion proteins into the endoplasmic reticulum and endo/lysosomal compartment, the TT-specific T(h)2 response was increased. The synthetic Pan DR epitope (PADRE) induced a stronger E7-specific response than the p30 epitope and its stimulatory effect was not limited to nuclear/cytoplasmic localization of the E7 antigen. These results suggest that in the optimization of immune responses by adding helper epitopes to DNA vaccines delivered by the gene gun, the cellular localization of the antigen needs to be taken into account.

• MeSH
• biolistika metody   MeSH
• buňky NIH 3T3 MeSH
• cytokiny metabolismus   MeSH
• DNA vakcíny aplikace a dávkování   farmakologie   MeSH
• endoplazmatické retikulum imunologie   metabolismus   MeSH
• HEK293 buňky MeSH
• lidé MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• Papillomavirus E7 - proteiny genetika   metabolismus   farmakologie   MeSH
• peptidové fragmenty genetika   farmakologie   MeSH
• plazmidy aplikace a dávkování   MeSH
• rekombinantní fúzní proteiny metabolismus   farmakologie   MeSH
• tetanový toxin genetika   farmakologie   MeSH
• vakcína proti malárii farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The CART (cocaine- and amphetamine-regulated transcript) peptide is an anorexigenic neuropeptide that acts in the hypothalamus. The receptor and the mechanism of action of this peptide are still unknown. In our previous study, we showed that the CART peptide binds specifically to PC12 rat pheochromocytoma cells in both the native and differentiated into neuronal phenotype. Two biologically active forms, CART(55-102) and CART(61-102), with equal biological activity, contain three disulfide bridges. To clarify the importance of each of these disulfide bridges in maintaining the biological activity of CART(61-102), an Ala scan at particular S-S bridges forming cysteines was performed, and analogs with only one or two disulfide bridges were synthesized. In this study, a stabilized CART(61-102) analog with norleucine instead of methionine at position 67 was also prepared and was found to bind to PC12 cells with an anorexigenic potency similar to that of CART(61-102). The binding study revealed that out of all analogs tested, [Ala(68,86)]CART(61-102), which contains two disulfide bridges (positions 74-94 and 88-101), preserved a high affinity to both native PC12 cells and those that had been differentiated into neurons. In food intake and behavioral tests with mice after intracerebroventricular administration, this analog showed strong and long-lasting anorexigenic potency. Therefore, the disulfide bridge between cysteines 68 and 86 in CART(61-102) can be omitted without a loss of biological activity, but the preservation of two other disulfide bridges and the full-length peptide are essential for biological activity.

• MeSH
• anorektika chemie   farmakologie   MeSH
• buňky PC12 MeSH
• cystin chemie   MeSH
• kompetitivní vazba MeSH
• krysa rodu rattus MeSH
• lokomoce účinky léků   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nocicepce účinky léků   MeSH
• peptidové fragmenty chemie   farmakologie   MeSH
• přijímání potravy účinky léků   MeSH
• proteiny nervové tkáně chemie   farmakologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Problematika srdečního selhání, zejména akutní dekompenzace, stále podněcuje k hledání a vývoji nových lékových skupin. V článku jsou uvedeny výsledky klinických studií, v nichž byly hodnoceny natriuretické peptidy, blokátory receptoru pro adenosin, blokátor receptorů pro vazopresin, relaxin a nová pozitivně inotropní látka – omecamtiv mecarbil. I když tyto nové lékové skupiny přinášejí klinické zlepšení, prognózu nemocných většina z nich nezlepšuje. Srdeční selhání je klinický syndrom charakterizovaný nedostatečnou srdeční činností, v jejímž důsledku dochází nejen ke zhoršení funkce řady orgánů, ale také k aktivaci mnoha humorálních působků a adaptačních mechanismů. Aktivuje se sympatikus, renin-angiotenzin-aldosteronový systém, vazopresin, endoteliny a cytokinový systém, které působí vesměs natrium-retenčně, vazokonstriktivně a proliferativně. Na druhé straně se aktivuje systém natriuretických peptidů, který působí kontraregulačně. Ve farmakoterapii srdečního selhání se uplatňují lékové skupiny, které potlačují tyto negativně působící mechanismy: inhibitory ACE, blokátory receptoru 1 pro angiotenzin II, betablokátory či blokátory aldosteronu. Při hledání dalších možností farmakoterapie srdečního selhání se objevují nové látky, které by mohly nepříznivě působící systémy blokovat: klinickým hodnocením procházely blokátory endotelinových receptorů či anticytokinové látky. Výsledky těchto studií však nebyly pozitivním přínosem pro klinickou praxi. Nicméně výzkum a vývoj nových látek v oblasti srdečního selhání, zejména akutního srdečního selhání, pokračuje dále. Další nedávno ukončené či probíhající studie s novými molekulami uvádí tab. 1 (která se zde bohužel nezobrazuje).

The issue of heart failure, mainly of acute decompensation, remains a challenge for the development of novel substances. This article presents the results of recent clinical trials, which evaluated natriuretic peptides, adenosine receptor blockers, vasopressin receptor blockers, relaxin and a new positive inotropic agent - omecamtiv mecarbil. Although these drugs lead to clinical improvement, the majority do not affect patient prognosis.

• Klíčová slova
• nesiritid, ularitid, rolofyllin, serelaxin, pozitivně inotropní látky, omecamtiv mecarbil, ASCEND - HF, SIRIUS I, SIRIUS II, EVEREST, PROTECT, RELAX - AHF, ATOMIC,
• MeSH
• antagonisté antidiuretického hormonu MeSH
• atriální natriuretický faktor farmakologie   terapeutické užití   MeSH
• benzazepiny farmakologie   terapeutické užití   MeSH
• diuretika farmakologie   terapeutické užití   MeSH
• dvojitá slepá metoda MeSH
• dyspnoe MeSH
• farmakoterapie * trendy   MeSH
• hodnocení léčiv MeSH
• hospitalizace MeSH
• kardiotonika terapeutické užití   MeSH
• klinické zkoušky jako téma MeSH
• lidé MeSH
• močovina analogy a deriváty   farmakologie   terapeutické užití   MeSH
• multicentrické studie jako téma MeSH
• natriuretické peptidy terapeutické užití   MeSH
• natriuretický peptid typu B farmakologie   terapeutické užití   MeSH
• peptidové fragmenty farmakologie   terapeutické užití   MeSH
• placeba MeSH
• randomizované kontrolované studie jako téma MeSH
• receptory vasopresinů terapeutické užití   MeSH
• rekombinantní proteiny farmakologie   terapeutické užití   MeSH
• relaxin farmakologie   terapeutické užití   MeSH
• srdeční selhání * farmakoterapie   mortalita   MeSH
• tolvaptan MeSH
• výsledek terapie MeSH
• xanthiny farmakologie   terapeutické užití   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH