Human IL-2 Antibody

Detection of Human IL-2 by Flow Cytometry

IL-2 induces IL-32 but TCR activation is required for IL-32 secretion. (A) IL-32 expression in CD3+ T from healthy donors (HDs) after 24 h of resting (unstimulated) either alone or in the presence of IFN-gamma & low (100 IU/ml) or high (3,000 IU/ml) amounts of IL-2 & after 24 h of either CD3 or CD3/CD28 stimulation determined by ICFC analysis. Representative data from n=2 independent experiments. (B–D) WB analyses of IL-32 expression in cell lysates & IL-32 secretion into the cell culture supernatants (enriched via Amicon Ultracentrifugal filters) by HD CD3+ T after treatment for 48 h with 500 or 3,000 IU/ml IL-2 alone (low or high IL-2, respectively) or treatment with high IL-2 for 48 h with subsequent CD3/CD28 stimulation for 72 (h) (B) Representative blot of lysates & supernatants (15 µg) from n=2 HDs (#1, #2) next to rIL-32 beta (4 ng, MW: 23.1 kDa) & rIL-32 gamma (4 ng, MW: 28.1 kDa). As a loading control, the housekeeping gene GAPDH (MW: 37 kDa) was detected on same blot. (C) Cumulative data of quantified IL-32 beta expression in cell lysates normalized to GAPDH for n=2–5 HDs, mean+SD, dots depict data from individual experiments, Student’s paired t-test, *p<0.05. (D) Cumulative data of quantified IL-32 beta secretion into culture supernatants displayed by the Area Under the Curve (AUC) for n=2–5 HDs, mean+SD. (E, F) IL-32 expression in HD T after resting (Unstim) or CD3/CD28 stimulation alone (Stim) or together with either an alphaIL-2 neutralizing antibody (Stim + alphaIL-2) or the respective isotype control (Stim + Isotype) for 24 (h) (E) Representative dot plots depicting IL32+CD25+ among live single T stimulated in the presence of the isotype or the alphaIL-2 antibody. (F) Cumulative data showing the frequency of IL-32+CD25+ among live single T , n=2 HDs, Student’s unpaired t-test, *p<0.05. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/39211051), licensed under a CC-BY license. Not internally tested by R&D Systems.