Among the complications observed after allogeneic hematopoietic stem cell transplantation, graft-versus-host disease (GVHD) is the primary cause of post-transplant mortality. The oral cavity is the second most affected organ target in chronic GVHD. Tissue damage results from the upregulation of inflammatory mediators, which play a critical role in the immunopathogenesis of the disease. This case series observational study aims to evaluate the participation of cytokines, chemokines, transcription factors, and heat shock proteins in the pathogenesis of oral GVHD (oGVHD), describing the mRNA expression of 28 genes selected. Peripheral blood mononuclear cells were isolated from six participants with oGVHD and two without GVHD, and relative expression of transcripts with established roles as inflammatory mediators was determined in triplicate using the human RT2 Profiler™ PCR Array. The gene expression levels in the group with oGVHD were mainly up-regulated compared to those without GVHD. PBMC from oGVDH expressed consistently higher IFN-γ, TNF, IL-1β, CCL2, HSP60 (HSPD1) and HSP90 (HSP90B1). These results can provide a basis for developing new molecular diagnostics and targets therapies for the clinical management of oGVHD.
Introduction:Early and reliable laboratory diagnosis is very important to guide treatment of and control strategies for a disease. Some technical problems can occur due to laboratory facilities and methodological procedures. Many in vitro diagnoses are based on colorimetric methods such as the quantitative Enzyme-Linked Immuno Sorbent Assay (ELISA), whereby optical density (OD) is compared with a standard curve. The readings of these assays are based on absorption by a spectrophotometric device of light from a narrow or broad spectrum of waves. Objective: The aim of this work was to read the ELISA plate using different spectrophotometry equipment. Methods: Human plasma samples from healthy volunteers (n=50) and volunteers with tuberculosis (n=24) were prepared previously by whole blood incubation in the commercial Interferon Gamma Release Assay (IGRA) for in vitro diagnosis of tuberculosis and then the Interferon ELISA was performed using commercial equipment from the same manufacturer. The ELISA micro plates were read in three spectrophotometers with different filters/wavelengths: 450, 630 and 450 nm. The optical densities were established and analyzed using the manufacturer's software program. The results of the readings performed at the recommended wavelength were compared with the other two filters/wavelengths. Results: Among the results obtained, 10 samples showed differences in the results of diagnosis. These discrepancies in metrology were associated with differences in the filters/wavelengths and were consequently identified as false results.
Conclusion:This shows the importance of following recommendations and the prior standardization of parameters for the required test validation. Many manufacturers must recommend and users must to follow recommendation of use of specific spectrum range to each protocol and validate the different instruments used to get desired quality, avoiding different and unexpected results.
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