Cytokine mRNA expression levels in resting and stimulated peripheral blood mononuclear cells (PBMCs)

Range Table - link
Organism Human Homo sapiens
Reference Bas A, Forsberg G, Hammarström S, Hammarström ML. Utility of the housekeeping genes 18S rRNA, beta-actin and glyceraldehyde-3-phosphate-dehydrogenase for normalization in real-time quantitative reverse transcriptase-polymerase chain reaction analysis of gene expression in human T lymphocytes. Scand J Immunol. 2004 Jun59(6):566-73 p.571 table 1PubMed ID15182252
Method P.567 left column 2nd paragraph: "Peripheral blood mononuclear cells (PBMCs) were stimulated by a polyclonal T-cell activator, and the cellular content of the three housekeeping genes was estimated by combination of cell counting and real-time qRTPCR determinations of b-actin mRNA, GAPDH mRNA and 18S rRNA levels. The cytokines interleukin-2 (IL-2), IL-4, IL-10, interferon-g (IFN-g), tumour necrosis factor-a (TNF-a) and transforming growth factor-b1 (TGF-b1) were chosen as model genes in the analysis of mRNA expression levels."
Comments P.571 left column 2nd paragraph: "The experimental set-up also allowed [investigators] to compare the average mRNA copy number per cell of the six cytokines in freshly isolated, resting PBMCs and in PBMCs after varying the time of T-cell activation. Generally, the cytokine mRNA levels were low in unstimulated PBMCs (<0.1 mRNA copies/cell, Table 1). TGF-β1 was an interesting exception with as much as 3.6 mRNA copies/cell in resting PBMCs and a poor response to activation (Table 1). IL-2, IFN-γ and TNF-α were all strongly induced in most blood donors (Table 1). However, the kinetics for IFN-γ induction differed from the other two cytokines. While IL-2 and TNF-α showed a maximum increase after 7 h of stimulation, IFN-γ reached maximum expression levels after 45 h (Fig. 5A-C, Table 1). The number of mRNA copies/cell was 10-fold higher for IFN-γ and TNF-α compared to IL-2, both in resting and in maximally stimulated PBMCs (Table 1). Two individuals had a T helper 2 profile with induction of IL-4 and only a marginal induction of IFN-γ and TNF-α mRNA. However, both IL-4 and IL-10 expression levels were still low after activation (<0.1 copies/cell, Table 1)." Note beneath table: "Average number of cytokine mRNA copies/cell in freshly isolated PBMCs and at time point of maximum response."
Entered by Uri M
ID 104228