The Candida albicans heat shock protein 90 kDa (hsp90-CA) is an important target for protective antibodies in disseminated candidiasis of experimental mice and humans. Hsp90-CA is present in the cell wall of Candida pseudohyphae or hyphae--typical pathogenic morphotypes in both mucosal and systemic Candida infections. However, the potential protective effects of hsp90-CA-specific antibodies in vaginal candidiasis has not yet been reported. In the present study we used various vaccine formulations (recombinant hsp90-CA protein and hsp90-CA-encoding DNA vaccine) and routes of administration (intradermal, intranasal, and intravenous) to induce both hsp90-CA-specific systemic and vaginal mucosa immune responses in experimental BALB/c mice. The results showed that intradermal recombinant hsp90-CA protein priming, followed by intranasal or intradermal recombinant hsp90-CA protein boosting induced significant increases in both serum and vaginal hsp90-CA-specific IgG and IgA antibodies compared to the control group, as well as enhanced hsp90-CA-specific splenocyte responses in vitro. In the intradermally boosted group, subsequent experimental vaginal Candida infection induced additional increases in the hsp90-CA specific IgG isotype, suggesting that Candida has the ability to induce a local hsp90-specific antibody (IgG) response during vulvovaginal candidiasis. Further work is required to elucidate the importance of immunity to highly conserved antigens during infection of the human female reproductive tract where a balance between immunity to and tolerance for commonly antigens such as hsp90 is necessary for the maintenance of fertility.

• MeSH
• aplikace intranazální MeSH
• Candida albicans imunologie   MeSH
• DNA vakcíny aplikace a dávkování   imunologie   MeSH
• financování organizované MeSH
• fungální proteiny imunologie   MeSH
• fungální vakcíny aplikace a dávkování   imunologie   MeSH
• injekce intradermální MeSH
• injekce intravenózní MeSH
• kandidóza vulvovaginální imunologie   MeSH
• lymfocyty imunologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• proliferace buněk MeSH
• proteiny tepelného šoku HSP90 imunologie   MeSH
• protilátky fungální analýza   krev   MeSH
• sekundární imunizace MeSH
• slezina imunologie   MeSH
• syntetické vakcíny aplikace a dávkování   imunologie   MeSH
• tvorba protilátek MeSH
• vagina imunologie   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH

The recombinant Outer surface protein A (rOspA) from Borrelia burgdorferi is a possible immunogen for protection of infected humans and animals against development of Lyme borreliosis (Lyme disease), a chronic tick-borne disease characterised by diverse dermatologic, neurologic, rheumatic, and cardiac manifestations. For several years, research and development have been directed towards a vaccine for the prevention of this debilitating disease. Numerous animal studies demonstrate that pre-existing antibodies against the outer surface proteins of B. burgdorferi can prevent infection and disease caused by this organism. In this communication, using recombinant DNA technology, genes from B. burgdorferi sensu stricto and B. afzelii were inserted into E. coli-expression vectors and the rOspA were produced. Our aim was to obtain rOspA protein in a purity and quantity desirable for immunization of experimental animals. rOspA is currently the most developed, molecularly-defined vaccine candidate for the prevention of Lyme borreliosis.

• MeSH
• antigeny povrchové biosyntéza   izolace a purifikace   MeSH
• bakteriální vakcíny biosyntéza   izolace a purifikace   MeSH
• Borrelia burgdorferi komplex imunologie   MeSH
• Borrelia burgdorferi imunologie   MeSH
• DNA vakcíny biosyntéza   izolace a purifikace   MeSH
• Escherichia coli MeSH
• genetické vektory MeSH
• lipoproteiny biosyntéza   izolace a purifikace   MeSH
• proteiny vnější bakteriální membrány biosyntéza   izolace a purifikace   MeSH
• vakcína proti lymeské nemoci biosyntéza   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of this work was isolation and purification of the major immunodominant protein, Outer surface protein C (OspC) of three members of the species group Borrelia burgdorferi, the causative agent of Lyme disease. Our aim was to obtain this protein in a quantity and purity sufficient for immunization of experimental animals. For optimalization of protein purification's yield we used immobilized metal ion affinity chromatography (IMAC) under different conditions. The greatest efficiency was achieved by using of HiTrap Chelating Column under native conditions.

• MeSH
• antigeny bakteriální biosyntéza   izolace a purifikace   MeSH
• DNA vakcíny biosyntéza   izolace a purifikace   MeSH
• Escherichia coli MeSH
• genetické vektory MeSH
• proteiny vnější bakteriální membrány biosyntéza   izolace a purifikace   MeSH
• vakcína proti lymeské nemoci biosyntéza   izolace a purifikace   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH