Comparison of mRNA and Protein Measures of Cytokines following Vaccination with Human Papillomavirus-16 L1 Virus-like Particles

Abstract

Background: mRNA expression signatures are frequently used as surrogate measures of cellular function and pathway changes. Few studies have directly compared results obtained using gene expression and multiplex protein assays for corresponding gene products.Methods: We used data available from a clinical trial of a human papillomavirus-16 vaccine that tracked gene expression and cytokine/chemokine production by peripheral blood mononuclear cells stimulated in culture with various antigens to evaluate the degree to which gene expression levels reflect observed levels of cytokines/chemokines. Twenty-six women enrolled in a phase II clinical trial of a human papillomavirus-16 vaccine were evaluated for gene expression (using the Affymetrix Human Genome Focus Array) and cytokine/chemokine levels (using a bead-based 22-plex cytokine assay developed by Linco Research, Inc.) before and after vaccination.Results: Our results suggest the presence of a wide range of correlations between mRNA expression and secreted protein levels. The strongest correlation was observed for IFN-γ (R = 0.90 overall levels; R = 0.69 when vaccine induced changes were evaluated). More modest overall correlations ranging from 0.40 to 0.80 were observed for MIP1A, IP10, TNF-α, MCP1, IL-2, GM-CSF, IL-5, RANTES, and IL-8. Weaker or no correlation was observed between gene expression and protein levels for the remaining cytokines/chemokines evaluated.Conclusion: The degree of correlation between gene expression and protein levels varied among different cytokines/chemokines. Impact: Researchers should be cautious when using mRNA expression array results as a proxy for protein levels using existing technologies. Cancer Epidemiol Biomarkers Prev; 19(4); 978-81. ©2010 AACR.

Salinity-induced activation of the myo-inositol biosynthesis pathway in tilapia gill epithelium

“…Inhibition of protein synthesis/translational efficiency during salinity stress would also explain the much higher levels of mRNA induction at 34 and 90 ppt compared with the magnitude of the corresponding protein increase. Stronger increases in mRNA compared with corresponding protein abundances in response to stress are not uncommon and have been observed for many other systems (Kaufmann and van Oudenaarden, 2007;Persson et al, 2009;Shebl et al, 2010;Schwanhäusser et al, 2011). Interestingly, two different MIPS mRNA variants are present in tilapia gill epithelium.…”

Upregulation of circulating cancer stem cell marker, DCLK1 but not Lgr5, in chemoradiotherapy-treated colorectal cancer patients

“…As a result, a CSC whose DCLK1 promoter has been hypermethylated and cannot express DCLK1 anymore may still carry DCLK1 proteins that have been built before hypermethylation. Several different studies confirmed that the mRNA level of a gene does not necessarily predict its protein level [68][69][70][71]. To this reason, the evaluation of DCLK1 protein instead of DCLK1 mRNA may result in more valuable information.…”

Calcitonin Gene-Related Peptide Modulates the Production of Pro-Inflammatory Cytokines Associated with Periprosthetic Osteolysis by THP-1 Macrophage-Like Cells

“…Similar reactions were observed with LPS-activated cells treated with CGRP for 48 h of incubation. These disparities between RNA and protein levels can be explained by a lack of correlation between gene expression and protein production and secretion [70,71,72]. Indeed, it has been detected that the expression and secretion profiles for cytokines in activated macrophages overlap but are temporally regulated [73].…”