Treg Depletion Using Anti-CD25 mAb in Irf8-/- Mice

To deplete Tregs, Irf8^-/- mice were treated i.p. with 1 mg of neutralizing anti-CD25 mAb (clone PC61, rat IgG1) or control rat IgG (isotype control) (Bio X Cell; West Lebanon, NH) on day -7 before infection, day 0, and day 7 p.i. [40 (link)]. The extent of Treg depletion was verified in MLN cells obtained from anti-CD25 mAb treated and isotype control mice on day 14 p.i. by flow cytometry. Single cell suspensions were prepared and the cells were surfaced stained with FITC-conjugated anti-CD4 mAb (clone GK1.5; eBioscience) and PE-conjugated anti-CD25 mAb (clone 7D4; eBiosciences), fixed, and permeabilized followed by intracellular staining with APC-conjugated anti-Foxp3 mAb (clone FJK-16s; eBioscience).

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Valanparambil R.M., Tam M., Gros P.P., Auger J.P., Segura M., Gros P., Jardim A., Geary T.G., Ozato K, & Stevenson M.M. (2017). IRF-8 regulates expansion of myeloid-derived suppressor cells and Foxp3+ regulatory T cells and modulates Th2 immune responses to gastrointestinal nematode infection. PLoS Pathogens, 13(10), e1006647.

Publication 2017

control rat IgG FITC-conjugated anti-CD4 mAb (clone GK1.5;

Cd25 Cells Clone Fitc Flow cytometry Igg1 Infection Intracellular Irf8 Isotype Mice

Corresponding Organization : Eunice Kennedy Shriver National Institute of Child Health and Human Development

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Variable analysis

independent variables

Treatment with anti-CD25 mAb (clone PC61, rat IgG1)
Treatment with control rat IgG (isotype control)

dependent variables

Extent of Treg depletion in MLN cells on day 14 post-infection, measured by flow cytometry

control variables

Mouse strain: Irf8-/- mice

controls

Positive control: Treatment with anti-CD25 mAb
Negative control: Treatment with isotype control rat IgG

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