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  • Title: Ubiquitous interleukin-1 alpha in fetal bovine serum may mislead the experimental results in vitro.
    Author: Hida S, Takeuchi A, Hayashi H, Yano T, Hopkins SJ, Onozaki K.
    Journal: Eur Cytokine Netw; 1995; 6(2):121-6. PubMed ID: 7578990.
    Abstract:
    Fetal bovine sera (FBS) from several commercial suppliers were fractionated by gel filtration. Interleukin-1 (IL-1) activity was bioassayed using the IL-1-specific murine T cell line D10(N4)M. All the sera examined contained IL-1-like activity, with molecular weights (M(r)) of 30 kDa and 15-10 kDa. Under isoelectric focusing (IEF), the majority of IL-1 activity in either 30 kDa or 15-10 kDa fractions was focused into a position of pl 5. The activity recovered from either IEF or gel filtration was inhibited by either recombinant human IL-1 receptor antagonist (rhlL-1ra) or by the antibody against human IL-1 alpha. These biological and physicochemical properties strongly suggest that the active molecules were bovine IL-1 alpha and its precursor. There was no correlation between the amount of endotoxin and IL-1 activity. Quantification of the fractionated IL-1 indicated its presence in concentrations of 200-5000 pg/ml equivalent to human IL-1. However, high levels of IL-1 were not apparent in unfractionated FBS. Proliferation of T cells in the presence of FBS absorbed with protein A-Sepharose was greater than that of cells in original FBS. Therefore, the activity in FBS as a whole appeared to result from the balance between IL-1 and the inhibitory molecule(s). These results suggest that data obtained from experiments conducted in the presence of FBS may be influenced by the effect of bovine IL-1.
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