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

New synthetic aminoxy lipids are designed and synthesized as building blocks for the formulation of functionalized nanoliposomes by microfluidization using a NanoAssemblr. Orthogonal binding of hyaluronic acid onto the outer surface of functionalized nanoliposomes via aminoxy coupling ( N-oxy ligation) is achieved at hemiacetal function of hyaluronic acid and the structure of hyaluronic acid-liposomes is visualized by transmission electron microscopy and cryotransmission electron microscopy. Observed structures are in a good correlation with data obtained by dynamic light scattering (size and ζ-potential). In vitro experiments on cell lines expressing CD44 receptors demonstrate selective internalization of fluorochrome-labeled hyaluronic acid-liposomes, while cells with down regulated CD44 receptor levels exhibit very low internalization of hyaluronic acid-liposomes. A method based on microfluidization mixing was developed for preparation of monodispersive unilamellar liposomes containing aminoxy lipids and orthogonal binding of hyaluronic acid onto the liposomal surface was demonstrated. These hyaluronic acid-liposomes represent a potentially new drug delivery platform for CD44-targeted anticancer drugs as well as for immunotherapeutics and vaccines.

• MeSH
• antigeny CD44 analýza   metabolismus   MeSH
• antitumorózní látky aplikace a dávkování   MeSH
• buněčné linie MeSH
• endocytóza MeSH
• fluorescenční barviva MeSH
• kyselina hyaluronová chemie   metabolismus   MeSH
• lidé MeSH
• lipidy chemická syntéza   MeSH
• liposomy chemie   terapeutické užití   MeSH
• mikrofluidika MeSH
• nádory farmakoterapie   MeSH
• systémy cílené aplikace léků metody   MeSH
• transmisní elektronová mikroskopie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem 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

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