Abstract
C-reactive protein (CRP), a prominent component of the innate immune system, is implicated in the pathophysiology of many conditions. CRP production primarily occurs in the liver; but contributions from other tissues is unclear. The Genotype-Tissue Expression Portal shows essentially no expression in whole blood and reports in the literature are conflicting. Multiple genomic variants influence serum levels of CRP. We measured CRP mRNA expression in leukocytes and sought to determine if rs1205 genotype influences leukocyte expression. Leukocytes were obtained from 20 women differing by genotype. Quantitative, real-time PCR (RT-qPCR) detected CRP and reference gene (GAPDH) mRNA. Leukocyte expression was calculated by the 2ΔCT method, and against a standard curve. Digital drop PCR was also used to calculate expression ratios. Student's t test and linear regression methods examined possible differences between genotypes. During 32 runs (10 replicates each), the RT-qPCR mean (SD) CRP/GAPDH ratio was 3.39 × 10–4 (SD 1.73 × 10–4) and 3.15 × 10–4 (SD 1.64 × 10–4) for TT and CC genotypes respectively, p = 0.76; and digital drop PCR results were similar. Serum CRP was not significantly different between genotypes, nor correlated with leukocyte expression. CRP is minimally expressed in unactivated leukocytes and this expression is not likely influenced by rs1205 genotype.
Highlights
C-reactive protein (CRP), a prominent component of the innate immune system, is implicated in the pathophysiology of many conditions
The present study was undertaken to determine if CRP expression could be detected in unactivated leukocytes from an American Indian population; and if it is influenced by rs1205 genotype
There is evidence of CRP mRNA expression by leukocytes[12], this is not uniformly accepted[9] and typically not examined in the unstimulated state[15].The ratio of CRP to reference gene expression in the present study was on the order of 1 0–4 by either of the four methods of calculation
Summary
C-reactive protein (CRP), a prominent component of the innate immune system, is implicated in the pathophysiology of many conditions. Quantitative, real-time PCR (RT-qPCR) detected CRP and reference gene (GAPDH) mRNA. Evidence for leukocyte expression of CRP has been contentious due to difficulties with non-specific antibody detection[12,13], differing real-time quantitative polymerase chain reaction (RT-qPCR) protocols, as well as characteristics of the CRP gene that complicate primer design for RT-qPCR. The present study was undertaken to determine if CRP expression could be detected in unactivated leukocytes from an American Indian population; and if it is influenced by rs1205 genotype. This variant is common, with a minor allele frequency of 46% in this p opulation[3]
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