Best L G, Azure C, Martell K, Tsosie K S, Voels B
University of North Dakota, Grand Forks, ND, USA.
Natural Sciences, Turtle Mountain Community College, Belcourt, ND, USA.
Sci Rep. 2021 Mar 11;11(1):5691. doi: 10.1038/s41598-021-85272-9.
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 2 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 (SD 1.73 × 10) and 3.15 × 10 (SD 1.64 × 10) 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.
C反应蛋白（CRP）是固有免疫系统的一个重要组成部分，与多种疾病的病理生理学有关。CRP主要在肝脏中产生，但其他组织的贡献尚不清楚。基因型-组织表达数据库显示全血中基本无表达，而文献报道相互矛盾。多个基因组变异影响血清CRP水平。我们测量了白细胞中CRP的mRNA表达，并试图确定rs1205基因型是否影响白细胞表达。从20名基因型不同的女性中获取白细胞。采用定量实时PCR（RT-qPCR）检测CRP和参照基因（GAPDH）的mRNA。通过2−ΔΔCT方法并参照标准曲线计算白细胞表达量。还使用数字液滴PCR计算表达比率。采用学生t检验和线性回归方法检验基因型之间可能存在的差异。在32次运行（每次10个重复）中，TT和CC基因型的RT-qPCR平均（标准差）CRP/GAPDH比率分别为3.39×10−3（标准差1.73×10−3）和3.15×10−3（标准差1.64×10−3），p = 0.76；数字液滴PCR结果相似。不同基因型之间血清CRP无显著差异，且与白细胞表达无相关性。CRP在未活化的白细胞中表达极少，且这种表达不太可能受rs1205基因型影响。
