Haemophilus parasuis (H. parasuis) is associated with meningitis, polyserositis, polyarthritis and bacterial pneumonia. At present, its prevention and control is difficult because of the lack of suitable subunit vaccines. Nowadays, high-throughput methods, immunoproteomics, are available to screen for more vaccine candidates. A protein extraction method for H. parasuis and two-dimensional electrophoresis (2-DE) were optimized to provide high-resolution profiles covering pH 3 to 10. Twenty immunoreactive spots were excised from gels after strict comparison between 2-DE Western blot membranes and the relevant gels. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) and MALDI-TOF-TOF-MS successfully identified 16 different proteins. Fifteen of them were reported as immunoreactive proteins in H. parasuis for the first time. In addition, recombinant HP5-7 (ABC transporter, periplasmic-binding protein) showed immunoreactivity both with hyperimmune rabbit serum and convalescent swine serum. Four recombinants of the 14 successfully expressed genes showed immunoreactivity with hyperimmune rabbit serum.

• MeSH
• 2D gelová elektroforéza MeSH
• bakteriální proteiny chemie   genetika   imunologie   MeSH
• Haemophilus parasuis chemie   klasifikace   genetika   imunologie   MeSH
• hemofilové infekce imunologie   mikrobiologie   veterinární   MeSH
• hmotnostní spektrometrie MeSH
• králíci MeSH
• molekulární sekvence - údaje MeSH
• nemoci prasat imunologie   mikrobiologie   MeSH
• prasata MeSH
• proteomika MeSH
• protilátky bakteriální analýza   imunologie   MeSH
• séroskupina MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Čína MeSH

Vascularized composite allografts (VCAs) of faces and extremities are subject to chronic rejection that is incompletely understood. Here we report on immunoproteomic evaluation of a full facial VCA removed 88 months after transplantation due to chronic rejection. CD8-positive T cells of donor (graft) origin infiltrate deep intragraft arteries in apposition to degenerating endothelium of chimeric recipient origin in association with arteriosclerotic alterations. Digital spatial proteomic profiling highlighted proteins expressed by activated cytotoxic T cells and macrophages as well as pathway components involved in atherogenic responses, including Indoleamine 2,3-Dioxygenase 1 (IDO1) and Stimulator of Interferon Response CGAMP Interactor (STING). Chronic facial VCA rejection thus involves T cell/macrophage-mediated accelerated arteriosclerosis not normally represented in punch biopsies and potentially driven by persistent graft-resident effector T cells and recipient target endothelium that chimerically repopulates graft arteries.

• MeSH
• přežívání štěpu MeSH
• proteomika MeSH
• rejekce štěpu etiologie   patologie   MeSH
• štěpy z kompozitní tkáně * transplantace   MeSH
• transplantace obličeje * MeSH
• vaskularizovaná kompozitní alotransplantace * MeSH
• Publikační typ
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Streptococcus suis is an important pathogen of pigs. In China, in addition to S. suis serotype 2, S. suis serotype 9 (SS9) is also a prevalent serotype. There is no vaccine available for SS9. An immunoproteome-based approach was developed to identify SS9 immunogenic proteins for vaccine development. Secreted proteins extracted from SS9 strain GZ0565 were screened by two-dimensional Western blotting using convalescent sera from pigs. Protein spots were excised from preparative gels and were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry, which led to the identification of ten immunogenic proteins (sortases, ABC transporter substrate-binding protein-maltose/maltodextrin, ABC transporter periplasmic protein, CHAP domain containing protein, peptidoglycan-binding LysM, elongation factor Tu, elongation factor G, thymidine kinase, molecular chaperone DnaK, hypothetical protein SSU98_2184). These novel immunogenic proteins, which are encoded by genes that are reasonably conserved among SS9 strains, may be developed as antigens for further study of SS9 vaccine.

• MeSH
• 2D gelová elektroforéza MeSH
• antigeny bakteriální chemie   imunologie   MeSH
• bakteriální proteiny chemie   imunologie   sekrece   MeSH
• nemoci prasat imunologie   mikrobiologie   prevence a kontrola   MeSH
• prasata MeSH
• proteomika metody   MeSH
• rekonvalescence MeSH
• sérum chemie   imunologie   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• Streptococcus suis genetika   imunologie   metabolismus   MeSH
• streptokokové infekce imunologie   mikrobiologie   prevence a kontrola   veterinární   MeSH
• streptokokové vakcíny biosyntéza   terapeutické užití   MeSH
• vakcinace MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Čína MeSH

Immunoproteomic analysis was applied to study the immunoreactivity of serum samples collected at different time points from a laboratory assistant accidentally infected with highly virulent strain of Francisella tularensis subsp. tularensis. Immunoblotting showed that the spectrum of F. tularensis antigens recognized specifically by immune sera remained with the exception for 1 antigen stable for up to 16 years after infection. Using immunoproteomics approach 10 immunoreactive antigens were successfully identified. Several new immunogenic F. tularensis proteins were described for the first time.

• MeSH
• 2D gelová elektroforéza metody   využití   MeSH
• finanční podpora výzkumu jako téma MeSH
• Francisella tularensis imunologie   patogenita   účinky léků   MeSH
• imunoblotting metody   využití   MeSH
• laboratorní infekce diagnóza   farmakoterapie   MeSH
• lidé MeSH
• protilátky bakteriální izolace a purifikace   MeSH
• spektrální analýza metody   využití   MeSH
• Check Tag
• lidé MeSH

Francisella tularensis is a Gram-negative, facultative intracellular bacterium causing disease in many mammalian species. The low infectious dose of F. tularensis and the ease of air-borne transmission are the main features responsible for the classification of this bacterium as a potential biological weapon. The live attenuated strain of F. tularensis live vaccine strain (LVS) is currently only effective vaccine against tularemia, however, this type of vaccine has not been approved for human use. In the presented study, sub-immunoproteome analysis was performed to search for new immunogenic proteins of Francisella tularensis LVS grown under different conditions. By this approach 35 immunoreactive antigens were identified, 19 of them showed to be novel immunogens. In conclusion, sub-immunoproteome analysis resulted in successful identification of novel immunoreactive proteins.

• MeSH
• 2D gelová elektroforéza MeSH
• antigeny bakteriální analýza   imunologie   izolace a purifikace   MeSH
• antisérum imunologie   MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• financování organizované MeSH
• Francisella tularensis imunologie   MeSH
• isoelektrická fokusace MeSH
• lidé MeSH
• membránové proteiny analýza   imunologie   izolace a purifikace   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• tandemová hmotnostní spektrometrie MeSH
• výpočetní biologie MeSH
• Check Tag
• lidé MeSH

Francisella tularensis subspecies tularensis is a highly virulent intracellular bacterial pathogen, causing the disease tularemia. However, a safe and effective vaccine for routine application against F. tularensis has not yet been developed. We have recently constructed the deletion mutants for the DsbA homolog protein (ΔdsbA/FSC200) and a hypothetical protein IglH (ΔiglH/FSC200) in the type B F. tularensis subsp. holarctica FSC200 strain, which exerted different protection capacity against parental virulent strain. In this study, we further investigated the immunological correlates for these different levels of protection provided by ΔdsbA/FSC200 and ΔiglH/FSC200 mutants. Our results show that ΔdsbA/FSC200 mutant, but not ΔiglH/FSC200 mutant, induces an early innate inflammatory response leading to strong Th1-like antibody response. Furthermore, vaccination with ΔdsbA/FSC200 mutant, but not with ΔiglH/FSC200, elicited protection against the subsequent challenge with type A SCHU S4 strain in mice. An immunoproteomic approach was used to map a spectrum of antigens targeted by Th1-like specific antibodies, and more than 80 bacterial antigens, including novel ones, were identified. Comparison of tularemic antigens recognized by the ΔdsbA/FSC200 post-vaccination and the SCHU S4 post-challenge sera then revealed the existence of 22 novel SCHU S4 specific antibody clones.

• MeSH
• atenuované vakcíny aplikace a dávkování   genetika   imunologie   MeSH
• bakteriální vakcíny aplikace a dávkování   genetika   imunologie   MeSH
• cytokiny sekrece   MeSH
• faktory virulence nedostatek   MeSH
• Francisella tularensis klasifikace   enzymologie   imunologie   MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední BALB C MeSH
• proteindisulfidisomerasy nedostatek   MeSH
• Th1 buňky imunologie   MeSH
• tularemie imunologie   prevence a kontrola   MeSH
• tvorba protilátek * MeSH
• zkřížená ochrana * MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• biologické markery * krev   MeSH
• časná diagnóza MeSH
• Crohnova nemoc * diagnóza   MeSH
• incidence MeSH
• klinické zkoušky jako téma MeSH
• kongresy jako téma MeSH
• Publikační typ
• zprávy MeSH