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A multidisciplinary approach to study a couple of monozygotic twins discordant for the chronic fatigue syndrome: a focus on potential salivary biomarkers

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By Federica Ciregia et al. • • October 10, 2013

Editor's Comment: Studies of discordant twins, that is pairs of twins in which one has an illness and the other doesn't, are extremely useful for uncovering both the effects and the pathogenesis of diseases. The reason these studies are so important is that they effectively rule out genetic factors. Once genetic factors are eliminated, the disease process itself can be studied. In this case, the researchers discovered that patients with ME/CFS expressed proteins in their saliva that were associated with inflammatory responses. The authors concluded that the saliva test could be used in conjunction with other proteomic (protein) tests to define a biomarker for the illness.

By Federica Ciregia et al.

Background: Chronic Fatigue Syndrome (CFS) is a severe, systemic illness characterized by persistent, debilitating and medically unexplained fatigue. The etiology and pathophysiology of CFS remains obscure, and diagnosis is formulated through the patient’s history and exclusion of other medical causes. Thereby, the availability of biomarkers for CFS could be useful for clinical research. In the present study, we used a proteomic approach to evaluate the global changes in the salivary profile in a couple of monozygotic twins who were discordant for CFS. The aim was to evaluate differences of salivary protein expression in the CFS patient in respect to his healthy twin.

Methods: Saliva samples were submitted to two-dimensional electrophoresis (2DE). The gels were stained with Sypro, and a comparison between CFS subject and the healthy one was performed by the software Progenesis Same Spot including the Analysis of variance (ANOVA test). The proteins spot found with a ?2-fold spot quantity change and p<0.05 were identified by Nano-liquid chromatography electrospray ionization tandem mass spectrometry. To validate the expression changes found with 2DE of 5 proteins (14-3-3 protein zeta/delta, cyclophilin A, Cystatin-C, Protein S100-A7, and zinc-alpha-2-glycoprotein), we used the western blot analysis. Moreover, proteins differentially expressed were functionally analyzed using the Ingenuity Pathways Analysis software with the aim to determine the predominant canonical pathways and the interaction network involved.

Results: The analysis of the protein profiles allowed us to find 13 proteins with a different expression in CFS in respect to control. Nine spots were up-regulated in CFS and 4 down-regulated. These proteins belong to different functional classes, such as inflammatory response, immune system and metabolism. In particular, as shown by the pathway analysis, the network built with our proteins highlights the involvement of inflammatory response in CFS pathogenesis.

Conclusions: This study shows the presence of differentially expressed proteins in the saliva of the couple of monozygotic twins discordant for CFS, probably related to the disease. Consequently, we believe the proteomic approach could be useful both to define a panel of potential diagnostic biomarkers and to shed new light on the comprehension of the pathogenetic pathways of CFS.

Source: Journal of Translational Medicine, October 2, 2013, 11:243 doi:10.1186/1479-5876-11-243. Federica Ciregia, Laura Giusti, Ylenia Da Valle, Elena Donadio, Arianna Consensi, Camillo Giacomelli, Francesca Sernissi, Pietro Scarpellini, Fabrizio Maggi, Antonio Lucacchini and Laura Bazzichi.

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