Autor(es): Hu Yuanqing,Shang Yuwei; Huang Jinlin; Wang Yan; Ren Fangzhe; Jiao Yang,Pan Zhiming; Jiao Xin-An

Resumo: Campylobacter jejuni is an important food-borne and zoonotic pathogen with a worldwide distribution. Humans and chickens are hosts of this pathogen. At present, there is no ideal vaccine for controlling human campylobacteriosis or the carriage of C. jejuni by chickens. Bacterial in vivo-induced antigens are useful as potential vaccine candidates and biomarkers of virulence. In this study, we developed a novel systematic immunoproteomics approach to identify in vivo-induced antigens among the total cell proteins of C. jejuni using pre-adsorbed sera from patients infected with C. jejuni. Overall, 14 immunoreactive spots were probed on a PVDF membrane using pre-adsorbed human sera against C. jejuni. Then, we excised these protein spots from a duplicate gel and identified using MALDI-TOF MS. In total, 14 in vivo-induced antigens were identified using PMF and BLAST analysis. The identified proteins include CadF (CadF-1 and CadF-2), CheW, TufB, DnaK, MetK, LpxB, HslU, DmsA, PorA, ProS, CJBH_0976, CSU_0396 and hypothetical protein cje135_05017. Real-time RT-PCR was performed on 9 genes to compare their expression levels in vivo and in vitro. The data showed that 8 of the 9 analyzed genes were significantly upregulated in vivo relative to in vitro. We successfully developed a novel immunoproteomics method for identifying in vivo-induced Campylobacter jejuni antigens by using pre-adsorbed sera from infected patients. This new analysis method may prove to be useful for identifying in vivo-induced antigens within any host infected by bacteria and will contribute to the development of new subunit vaccines.

Palavras-Chave: 2-DE, C. jejuni, CCDA, Campy blood-free selective medium, Campylobacter jejuni, DFI, DOC, GBS, Guillain-Barré syndrome, HRP, IEF, IVET, IVIAT, Immunoproteomics, In vivo-induced antigen, LOS, MALDI-TOF MS, MBP, NMRC, Naval Medical Research Center, PMF, STM, differential fluorescence induction, horseradish peroxidase, in vivo expression technology, in vivo-induced antigen technology, isoelectric focusing, lipooligosaccharides, maltose-binding protein, matrix-assisted laser desorption ionization-time of flight mass spectrometry, peptide mass fingerprinting, signature-tagged mutagenesis, sodium deoxycholate, two-dimensional echocardiography

Imprenta: Biochimica et Biophysica Acta, v. 1830, n. 11, p. 5229-5235, 2013

Identificador do objeto digital: 10.1016/j.bbagen.2013.06.042

Descritores: Guillain-Barre Syndrome - Biosynthesis ; Guillain-Barre Syndrome - Cell ; Guillain-Barre Syndrome - Pathogenesis ; Guillain-Barre Syndrome - Proteins ; Guillain-Barre Syndrome - Antibodies ; Guillain-Barre Syndrome - Infectious diseases ; Guillain-Barre Syndrome - Real Time PCR ; Guillain-Barre Syndrome - Vaccine ; Guillain-Barre Syndrome - Immunology

Data de publicação: 2013