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
One of the proposed strategies for the development of a more efficient HIV-1 vaccine is based on the identification of proteins binding to a paratope of chosen broadly neutralizing antibody (bNAb) that will mimic cognate HIV-1 Env (glyco)protein epitope and could be used as potent immunogens for induction of protective virus-neutralizing antibodies in the immunized individuals. To verify this "non-cognate ligand" concept, we developed a highly complex combinatorial library designed on a scaffold of human myomesin-1 protein domain and selected proteins called Myomedins specifically binding to variable regions of HIV-1 broadly neutralizing antibody 10E8. Immunization of mice with these Myomedin variants elicited the production of HIV-1 Env-specific antibodies. Hyperimmune sera bound to Env pseudotyped viruses and weakly/moderately neutralized 54% of tested clade A, B, C, and AE pseudotyped viruses variants in vitro. These results demonstrate that Myomedin variants have the potential to mimic Env epitopes and could be used as potential HIV-1 vaccine components.
- MeSH
- epitopy MeSH
- genové produkty env - virus lidské imunodeficience genetika MeSH
- HIV infekce * prevence a kontrola MeSH
- HIV protilátky MeSH
- HIV-1 * genetika MeSH
- myši MeSH
- neutralizující protilátky MeSH
- pseudotypování virů MeSH
- široce neutralizující protilátky MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Gadolinium (Gd)-based contrast agents are extensively used for magnetic resonance imaging (MRI). Liposomes are potential nanocarrier-based biocompatible platforms for development of new generations of MRI diagnostics. Liposomes with Gd-complexes (Gd-lip) co-encapsulated with thrombolytic agents can serve both for imaging and treatment of various pathological states including stroke. In this study, we evaluated nanosafety of Gd-lip containing PE-DTPA chelating Gd+3 prepared by lipid film hydration method. We detected no cytotoxicity of Gd-lip in human liver cells including cancer HepG2, progenitor (non-differentiated) HepaRG, and differentiated HepaRG cells. Furthermore, no potential side effects of Gd-lip were found using a complex system including general biomarkers of toxicity, such as induction of early response genes, oxidative, heat shock and endoplasmic reticulum stress, DNA damage responses, induction of xenobiotic metabolizing enzymes, and changes in sphingolipid metabolism in differentiated HepaRG. Moreover, Gd-lip did not show pro-inflammatory effects, as assessed in an assay based on activation of inflammasome NLRP3 in a model of human macrophages, and release of eicosanoids from HepaRG cells. In conclusion, this in vitro study indicates potential in vivo safety of Gd-lip with respect to hepatotoxicity and immunopathology caused by inflammation.
- MeSH
- diethylentriaminpentaacetát gadolinia * škodlivé účinky toxicita MeSH
- fibrinolytika MeSH
- fosfatidylethanolaminy * škodlivé účinky toxicita MeSH
- hepatocyty účinky léků MeSH
- inflamasomy MeSH
- kontrastní látky * MeSH
- kultivované buňky MeSH
- lidé MeSH
- liposomy * MeSH
- magnetická rezonanční tomografie * MeSH
- makrofágy účinky léků MeSH
- nanočástice MeSH
- nosiče léků * MeSH
- protein NLRP3 MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
New synthetic aminooxy lipid was designed and synthesized as a building block for the formulation of functionalised nanoliposomes (presenting onto the outer surface of aminooxy groups) by microfluidic mixing. Orthogonal binding of cellular mannan (Candida glabrata (CCY 26-20-1) onto the outer surface of functionalised nanoliposomes was modified by orthogonal binding of reducing termini of mannans to oxime lipids via a click chemistry reaction based on aminooxy coupling (oxime ligation). The aminooxy lipid was proved as a suitable active component for preparation of functionalised nanoliposomes by the microfluidic mixing method performed with the instrument NanoAssemblrTM. This "on-chip technology" can be easily scaled-up. The structure of mannan-liposomes was visualized by transmission and scanning electron microscopy, including immunogold staining of recombinant mannan receptor bound onto mannosylated-liposomes. The observed structures are in a good correlation with data obtained by DLS, NTA, and TPRS methods. In vitro experiments on human and mouse dendritic cells demonstrate selective internalisation of fluorochrome-labelled mannan-liposomes and their ability to stimulate DC comparable to lipopolysaccharide. We describe a potentially new drug delivery platform for mannan receptor-targeted antimicrobial drugs as well as for immunotherapeutics. Furthermore, the platform based on mannans bound orthogonally onto the surface of nanoliposomes represents a self-adjuvanted carrier for construction of liposome-based recombinant vaccines for both systemic and mucosal routes of administration.
- MeSH
- adjuvancia imunologická farmakologie MeSH
- antigeny povrchové metabolismus MeSH
- Candida glabrata chemie MeSH
- dendritické buňky imunologie MeSH
- hydroxylaminy chemická syntéza chemie MeSH
- lektiny typu C imunologie MeSH
- lektiny vázající mannosu imunologie MeSH
- lidé MeSH
- lipidy chemická syntéza chemie MeSH
- liposomy chemie imunologie farmakologie MeSH
- mannany chemie imunologie farmakologie MeSH
- mikrofluidika metody MeSH
- myši inbrední BALB C MeSH
- nanočástice chemie MeSH
- receptory buněčného povrchu imunologie MeSH
- syntetická chemie okamžité shody MeSH
- velikost částic MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Fatty acyl analogues of muramyldipeptide (MDP) (abbreviated N-L18 norAbuGMDP, N-B30 norAbuGMDP, norAbuMDP-Lys(L18), norAbuMDP-Lys(B30), norAbuGMDP-Lys(L18), norAbuGMDP-Lys(B30), B30 norAbuMDP, L18 norAbuMDP) are designed and synthesized comprising the normuramyl-l-α-aminobutanoyl (norAbu) structural moiety. All new analogues show depressed pyrogenicity in both free (micellar) state and in liposomal formulations when tested in rabbits in vivo (sc and iv application). New analogues are also shown to be selective activators of NOD2 and NLRP3 (inflammasome) in vitro but not NOD1. Potencies of NOD2 and NLRP3 stimulation are found comparable with free MDP and other positive controls. Analogues are also demonstrated to be effective in stimulating cellular proliferation when the sera from mice are injected sc with individual liposome-loaded analogues, causing proliferation of bone marrow-derived GM-progenitors cells. Importantly, vaccination nanoparticles prepared from metallochelation liposomes, His-tagged antigen rOspA from Borrelia burgdorferi, and lipophilic analogue norAbuMDP-Lys(B30) as adjuvant, are shown to provoke OspA-specific antibody responses with a strong Th1-bias (dominance of IgG2a response). In contrast, the adjuvant effects of Alum or parent MDP show a strong Th2-bias (dominance of IgG1 response).
- MeSH
- acetylmuramyl-alanyl-isoglutamin analogy a deriváty chemie imunologie farmakologie MeSH
- adjuvancia imunologická chemie farmakologie MeSH
- antigeny povrchové chemie imunologie farmakologie MeSH
- bakteriální vakcíny chemie imunologie farmakologie MeSH
- Borrelia burgdorferi imunologie MeSH
- HEK293 buňky MeSH
- imunizace MeSH
- lidé MeSH
- lipoproteiny chemie imunologie farmakologie MeSH
- lymeská nemoc imunologie mikrobiologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- protein NLRP3 agonisté imunologie MeSH
- proteiny vnější bakteriální membrány chemie imunologie farmakologie MeSH
- RAW 264.7 buňky MeSH
- tvorba protilátek MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Several plasminogen activators (PAs) have been found effective in treating different thromboembolic diseases. However, administration of conventional thrombolytic therapy is limited by a low efficacy of present formulations of PAs. Conventional treatments using these therapeutic proteins are associated with several limitations including rapid inactivation and clearance, short half-life, bleeding complications or non-specific tissue targeting. Liposome-based formulations of PAs such as streptokinase, tissue-plasminogen activator and urokinase have been developed to improve the therapeutic efficacy of these proteins. Resulting liposomal formulations were found to preserve the original activity of PAs, promote their selective delivery and improve thrombus targeting. Therapeutic potential of these liposome-based PAs has been demonstrated successfully in various pre-clinical models in vivo. Reductions in unwanted side effects (e.g., hemorrhage or immunogenicity) as well as enhancements of efficacy and safety were achieved in comparison to currently existing treatment options based on conventional formulations of PAs. This review summarizes present achievements in: (i) preparation of liposome-based formulations of various PAs, (ii) development of PEGylated and targeted liposomal PAs, (iii) physico-chemical characterization of these developed systems, and (iv) testing of their thrombolytic efficacy. We also look to the future and the imminent arrival of theranostic liposomal formulations to move this field forward.
- MeSH
- aktivátor plasminogenu urokinasového typu aplikace a dávkování terapeutické užití MeSH
- fibrinolytika aplikace a dávkování terapeutické užití MeSH
- lidé MeSH
- liposomy chemie ultrastruktura MeSH
- metaloendopeptidasy aplikace a dávkování terapeutické užití MeSH
- nanostruktury chemie ultrastruktura MeSH
- plasmin aplikace a dávkování terapeutické užití MeSH
- streptokinasa aplikace a dávkování terapeutické užití MeSH
- tkáňový aktivátor plazminogenu aplikace a dávkování terapeutické užití MeSH
- tromboembolie farmakoterapie MeSH
- trombolytická terapie metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Lyme disease, Borrelia burgdorferi-caused infection, if not recognized and appropriately treated by antibiotics, may lead to chronic complications, thus stressing the need for protective vaccine development. The immune protection is mediated by phagocytic cells and by Borrelia-specific complement-activating antibodies, associated with the Th1 immune response. Surface antigen OspC is involved in Borrelia spreading through the host body. Previously we reported that recombinant histidine tagged (His-tag) OspC (rOspC) could be attached onto liposome surfaces by metallochelation. Here we report that levels of OspC-specific antibodies vary substantially depending upon whether rOspC possesses an N' or C' terminal His-tag. This is the case in mice immunized: (a) with rOspC proteoliposomes containing adjuvants MPLA or non-pyrogenic MDP analogue MT06; (b) with free rOspC and Montanide PET GEL A; (c) with free rOspC and alum; or (d) with adjuvant-free rOspC. Stronger responses are noted with all N'-terminal His-tag rOspC formulations. OspC-specific Th1-type antibodies predominate post-immunization with rOspC proteoliposomes formulated with MPLA or MT06 adjuvants. Further analyses confirmed that the structural features of soluble N' and C' terminal His-tag rOspC and respective rOspC proteoliposomes are similar including their thermal stabilities at physiological temperatures. On the other hand, a change in the position of the rOspC His-tag from N' to C' terminal appears to affect substantially the immunogenicity of rOspC arguably due to steric hindrance of OspC epitopes by the C' terminal His-tag itself and not due to differences in overall conformations induced by changes in the His-tag position in rOspC variants.
- MeSH
- adjuvancia imunologická * MeSH
- antigeny bakteriální aplikace a dávkování chemie imunologie MeSH
- Borrelia burgdorferi imunologie MeSH
- ELISA MeSH
- imunizace MeSH
- lymeská nemoc imunologie MeSH
- modely u zvířat MeSH
- myši MeSH
- proteiny vnější bakteriální membrány aplikace a dávkování chemie imunologie MeSH
- proteolipidy MeSH
- protilátky bakteriální imunologie MeSH
- rekombinantní fúzní proteiny aplikace a dávkování chemie imunologie izolace a purifikace MeSH
- sekundární struktura proteinů MeSH
- specificita protilátek imunologie MeSH
- stabilita proteinů MeSH
- tvorba protilátek imunologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Pro-apoptotic analogues of vitamin E (VE) exert selective anti-cancer effect on various animal cancer models. Neither suitable formulation of α-tocopheryl succinate (α-TOS), representative semi-synthetic VE analogue ester, nor suitable formulations of the other VE analogues for clinical application have been reported yet. The major factor limiting the use of VE analogues is their low solubility in aqueous solvents. Due to the hydrophobic character of VE analogues, liposomes are predetermined as suitable delivery system. Liposomal formulation prevents undesirable side effects of the drug, enhances the drug biocompatibility, and improves the drug therapeutic index. Liposomal formulations of VE analogues especially of α-TOS and α-tocopheryl ether linked acetic acid (α-TEA) have been developed. The anti-cancer effect of these liposomal VE analogues has been successfully demonstrated in pre-clinical models in vivo. Present achievements in: (i) preparation of liposomal formulations of VE analogues, (ii) physico-chemical characterization of these developed systems and (iii) testing of their biological activity such as induction of apoptosis and evaluation of anti-cancer effect are discussed in this review.
- MeSH
- alfa-tokoferol aplikace a dávkování MeSH
- antitumorózní látky aplikace a dávkování chemie MeSH
- apoptóza účinky léků MeSH
- chemie farmaceutická MeSH
- lidé MeSH
- lipidy chemie MeSH
- liposomy MeSH
- nádory farmakoterapie patologie MeSH
- rozpustnost MeSH
- systémy cílené aplikace léků metody MeSH
- vitamin E aplikace a dávkování analogy a deriváty chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Lyme disease caused by spirochete Borrelia burgdorferi sensu lato, is a tick-born illness. If the infection is not eliminated by the host immune system and/or antibiotics, it may further disseminate and cause severe chronic complications. The immune response to Borrelia is mediated by phagocytic cells and by Borrelia-specific complement-activating antibodies associated with Th1 cell activation. A new experimental vaccine was constructed using non-lipidized form of recombinant B. burgdorferi s.s. OspC protein was anchored by metallochelating bond onto the surface of nanoliposomes containing novel nonpyrogenic lipophilized norAbuMDP analogues denoted MT05 and MT06. After i.d. immunization, the experimental vaccines surpassed Alum with respect to OspC-specific titers of IgG2a, IgG2b isotypes when MT06 was used and IgG3, IgM isotypes when MT05 was used. Both adjuvants exerted a high adjuvant effect comparable or better than MDP and proved themselves as nonpyrogenic.
- MeSH
- acetylmuramyl-alanyl-isoglutamin chemie toxicita MeSH
- antigeny bakteriální imunologie MeSH
- Borrelia burgdorferi imunologie MeSH
- chelátory chemie toxicita MeSH
- diferenciální skenovací kalorimetrie MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- ELISA MeSH
- liposomy MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- nanočástice chemie toxicita MeSH
- nosiče léků chemie toxicita MeSH
- proteiny vnější bakteriální membrány imunologie MeSH
- protilátky bakteriální krev MeSH
- radiační rozptyl MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- světlo MeSH
- transmisní elektronová mikroskopie MeSH
- vakcína proti lymeské nemoci aplikace a dávkování imunologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We designed and synthesised a series of new cationic lipids based on spermine linked to various hydrophobic anchors. These lipids could be potentially useful for the preparation of stable cationic liposomes intended for the construction of drug targeting systems applicable in the field of anticancer/antiviral therapy, vaccine carriers, and vectors for the gene therapy. Low in vitro toxicity was found for these compounds, especially for LD1, in several cell lines. The delivery of both a fluorescence marker (calcein) and antiviral drugs into cells has been achieved owing to a large extent of internalization of cationic liposomes (labelled by Lyssamine-Rhodamine PE or fluorescein-PE) as demonstrated by fluorescent microscopy and quantified by flow cytometry. The bovine herpes virus type 1 (BHV-1) virus infection in vitro model using MDBK cells was employed to study the effect of the established antiviral drug HPMPC (Cidofovir®) developed by Prof. A. Holý. Inhibition of BHV-1 virus replication was studied by quantitative RT-PCR and confirmed by both Hoffman modulation contrast microscopy and transmission electron microscopy. We found that in vitro antiviral activity of HPMPC was significantly improved by formulation in cationic liposomes, which decreased the viral replication by about 2 orders of magnitude.
- MeSH
- antivirové látky aplikace a dávkování farmakologie MeSH
- bovinní herpesvirus 1 účinky léků fyziologie MeSH
- buněčné kultury MeSH
- buněčné linie MeSH
- cytopatogenní efekt virový MeSH
- cytosin aplikace a dávkování analogy a deriváty farmakologie MeSH
- fluorescenční mikroskopie MeSH
- kationty MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- ledviny cytologie virologie MeSH
- lipidy chemie MeSH
- liposomy MeSH
- nosiče léků chemie MeSH
- organofosfonáty aplikace a dávkování farmakologie MeSH
- replikace viru účinky léků MeSH
- skot MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
