Immunoproteomic identification of antigenic candidate Campylobacter jejuni and human peripheral nerve proteins involved in Guillain-Barré syndrome

Immunoproteomics is become a potent methodology used for identifying immunoreactive proteins. In this study, an immunoproteomic approach based on 2-dimensional gel electrophoresis (2D-PAGE) and immunoblotting combined with high resolution mass spectrometry (MS) was used to identify immunoreactive proteins that might be involved in mechanisms of Guillain-Barr\uc3\ua9 syndrome (GBS) development, regardless of their potential reciprocal molecular mimicry. Proteins isolated from C. jejuni and human peripheral nerve tissue (HPN) were separated with 2D SDS-PAGE and subjected to western blotting using serum samples from GBS patients. The peptides generated after proteolysis of the immunoreactive proteins were submitted to nanoflow-high performance liquid chromatography-nano electrospray ionization coupled to high resolution mass spectrometry (nHPLC-nESI-MS and MS/MS) followed by SEQUESTdata analysis for proteins identification. In C. jejuni, immunoreactivity was found for GroEL and DnaK, structural proteins (MOMP), key enzymatic proteins necessary for the microbial proliferation (adenylate kinase, enolase, inorganic pyrophosphatase and aspartate ammonia-lyase), and antioxidant enzymes (alkyl hydroperoxide reductase-AhpC and DNA protection during starvation protein - DNA protection factor against Fe2+-mediated oxidative stress).HPN immunoreactive proteins identified were heat shock proteins (HSP), intermediate filaments (vimentin and desmin), and other proteins and enzymes such as troponin/tropomyosin complex and ATP synthase subunit beta and the keratan sulfate proteoglycan lumican. The targeting of vimentin and desmin, suggested that the neuronal autoimmune damage is specifically directed to intermediate neuronal (vimentin) and neuromuscular IF, probably localized nearby cell surface, affording increased accessibility to autoantibodies. These findings suggest that the post-infectious development of GBS may be also associated to additional concomitant immune factors that lead to nerve damage generated by auto-immune trigger(s) different from molecular mimicry

Guillain Barré syndrome (GBS) : new insights in the molecular mimicry between C. jejuni and human peripheral nerve (HPN) proteins

Profile and immunoreactivity of proteins from HPN tissue, and from Campylobacter jejuni (O:19) were investigated. Proteins were extracted, separated by SDS-PAGE, their cross reactivity monitored by Western blotting, and identified by nHPLC-nESI-HRMS analysis. Proteins from C. jejuni, at Mw ~70KDa were chaperone/co-chaperone proteins (GroEL, DnaK and HtpG). In the corresponding HPN band were serum albumin, neurofilament light peptide, cytoskeletal keratins and one HSP 70 and one HSP60. These chaperones reciprocally share high primary sequence homology and conservation of their known epitopes. These findings suggest that HSP chaperones may be suitable candidates involved in the molecular mimicry triggering GBS

Myelin-derived and putative molecular mimic peptides share structural properties in aqueous and membrane-like environments

Abstract Background: Despite intense research, the causes of various neurological diseases remain enigmatic to date. A role for viral or bacterial infection and associated molecular mimicry has frequently been suggested in the etiology of neurological diseases, including demyelinating autoimmune disorders, such as multiple sclerosis. Pathogen mimics of myelin-derived autoimmune peptides have been described in the literature and shown to induce myelin autoimmune responses in animal models. Methods: We carried out a structural study on myelin-derived peptides, and mimics thereof from various pathogens, in aqueous and membrane-like environments, using conventional and synchrotron radiation circular dichroism spectroscopy. A total of 13 peptides from the literature were studied, and 290 circular dichroism spectra were analysed. In addition, peptide structure predictions and vesicle aggregation assays were performed. Results: The results indicate a high level of similarity in the biophysical and folding properties of the peptides from either myelin proteins or proteins from pathogenic viruses or bacteria; essentially all of the studied peptides folded in the presence of lipid vesicles or under other membrane-mimicking conditions, which is a sign of membrane interaction. Many of the peptides presented remarkable similarities in their conformation in different environments. Conclusions: As most of the studied epitope segments in myelin proteins are associated with membrane-binding sites, our results support a view of molecular mimicry, involving lipid membrane interaction propensity and similar conformational properties, possibly playing a role in demyelinating disease. The results suggest mechanisms related to protein amphiphilicity and order-disorder transitions in the recognition of peptide epitopes in autoimmune demyelination