BB0405 is a surface exposed Borrelia burgdorferi protein and its vaccination protected mice against B. burgdorferi infection. As BB0405 is highly conserved across different B. burgdorferi sensu lato species, we investigated whether vaccination with recombinant BB0405 or through intradermal bb0405 DNA tattoo vaccination could provide protection against different Borrelia species, specifically against Borrelia afzelii, the predominant B. burgdorferi sensu lato genospecies causing Lyme borreliosis across Eurasia. We immunized C3H/HeN mice with recombinant BB0405 or with a codon-optimized bb0405 DNA vaccine using the pVAC plasmid and immunized corresponding control groups mice with only adjuvant or empty vectors. We subsequently subjected these immunized mice to a tick challenge with B. afzelii CB43-infected Ixodes ricinus nymphs. Upon vaccination, recombinant BB0405 induced a high total IgG response, but bb0405 DNA vaccination did not elicit antibody responses. Both vaccine formulations did not provide protection against Borrelia afzelii strain CB43 after tick challenge. In an attempt to understand the lack of protection of the recombinant vaccine, we determined expression of BB0405 and showed that B. afzelii CB43 spirochetes significantly and drastically downregulate the expression of BB0405 protein at 37 °C compared to 33 °C, where as in B. burgdorferi B31 spirochetes expression levels remain unaltered. Vaccination with recombinant BB0405 was previously shown to protect against B. burgdorferi sensu stricto. Here we show that vaccination with either recombinant BB0405 (or non-immunogenic bb0405 DNA), despite being highly conserved among B. burgdorferi sl genospecies, does not provide cross-protection against B. afzelii, mostly likely due to downregulation of this protein in B. afzelii in the mammalian host.
- MeSH
- Borrelia burgdorferi komplex imunologie MeSH
- Borrelia burgdorferi imunologie MeSH
- DNA vakcíny imunologie terapeutické užití MeSH
- imunogenicita vakcíny MeSH
- lymeská nemoc imunologie prevence a kontrola MeSH
- myši inbrední C3H MeSH
- myši MeSH
- proteiny vnější bakteriální membrány imunologie terapeutické užití MeSH
- rekombinantní proteiny imunologie terapeutické užití MeSH
- tvorba protilátek MeSH
- vakcína proti lymeské nemoci imunologie terapeutické užití MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem 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
Lymeská borelióza je vážným onemocněním, jehož původcem je v Evropě nejčastěji Borrelia garinii. Pro pochopení patogeneze tohoto onemocnění je důležité získat informace o proteinovém složení původce a významným zdrojem těchto informací je proteomová analýza. Pro identifikaci pomocí dvojrozměrové elektroforézy a hmotnostní spektrometrie bylo vybráno 63 proteinových spot Borrelia garinii (kmen CB61). Z nich bylo úspěšně identifikováno 40, většina se však vyskytovala v různých izoformách. Identifikováno tak bylo celkem 16 různých proteinů. Převažují proteiny s enzymatickou funkcí, ale podařilo se také identifikovat některé proteiny zahrnuté v enzootickém cyklu. Jsou to zejména proteiny skupiny Osp, membránový protein p66 a flagelární protein.
Lyme disease is a serious illness caused in Europe most frequently by Borrelia garinii. It is important to find information about protein content for understanding pathogenesis of Lyme disease. Proteome analysis is an important source of this information. We used 2-dimensional electrophoresis and mass spectrometry for identification of 63 chosen Borrelia garinii (strain CB61) protein spots. Overall 40 protein spots were identified however most of them occurred in several isoforms. Totally 16 different proteins were identified. Among them we identified several enzymes. Proteins that participate in enzootic cycle of pathogen are more interesting. They are mainly proteins of Osp group, membrane protein p66 and flagellar protein.
- MeSH
- 2D gelová elektroforéza metody využití MeSH
- biologické markery MeSH
- Borrelia burgdorferi komplex genetika izolace a purifikace MeSH
- hmotnostní spektrometrie metody využití MeSH
- klíště mikrobiologie parazitologie patogenita MeSH
- lidé MeSH
- lymeská nemoc genetika imunologie mikrobiologie MeSH
- proteomika metody trendy MeSH
- vakcína proti lymeské nemoci genetika imunologie MeSH
- virulence genetika imunologie MeSH
- Check Tag
- lidé MeSH
Verification of the efficacy of Biocan B inj. ad us. vet. (Bioveta, a.s.) was done by challenge testing. Ticks collected in the nature were used as natural vectors of the infection. Six beagles and two control ones were used in the test. Formation of outer surface protein A specific antibodies (OspA antibodies) and borrelia specific immonoglobulins (IgG) was measured by Western blot and EIA in the sera samples. The tissue samples were used for detection of borreliae by cultivation method and dark field microscopy (DFM). Formation of IgG antibodies and OspA antibodies after vaccination was observed. The maximum titer level of antibodies was reached between 21. and 49. day after vaccination and then slowly decreased. Presence of borreliae was detected only in skin biopsies of non-vaccinated dogs. The post mortem tissue samples showed presence of borreliae in all of the samples of the non-vaccinated dogs. The tissues of the vaccinated dogs were not infected with borreliae, except for two samples of dog with low titer levels of OspA antibodies. The development of the new vaccine is based on preparation of recombinant outer surface proteins (e.g. rOspA and rOspC) of B. afzelii, B. burgdorferi and B. garinii origin. Chosen recombinant proteins were successfully expressed in E. coli. The obtained purified proteins are currently being tested on laboratory BALB/c mice. Formation of specific antibodies against some recombinant proteins has been confirmed. These proteins are suitable candidates for preparation of a vaccine prototype and they will be subsequently used in challenge tests.
- MeSH
- antigeny povrchové imunologie MeSH
- bakteriální vakcíny imunologie MeSH
- Borrelia burgdorferi komplex imunologie MeSH
- infekce bakteriemi rodu Borrelia prevence a kontrola veterinární MeSH
- lipoproteiny imunologie MeSH
- nemoci psů imunologie prevence a kontrola MeSH
- proteiny vnější bakteriální membrány imunologie MeSH
- protilátky bakteriální krev MeSH
- psi MeSH
- syntetické vakcíny imunologie MeSH
- vakcína proti lymeské nemoci imunologie MeSH
- vakcinace MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
