Mice were injected i.p. with 300 μg isotype control or blocking antibodies purchased from BioXCell (West Lebanon, NH): rat IgG2a (clone: 2A3) – control for anti-PD-L2, rat IgG2b (clone: LTF-2) – control for anti-PD-L1, rat anti-PD-L1 (clone: 10F.9G2), rat anti-PD-L2 (clone: TY25). The antibodies were administered to the mice 24 h prior to IRI. Tregs were isolated from spleen of naïve WT mice with Dynal (Carlsbad, CA) CD4 negative selection kit and Miltenyi Biotec (Auburn, CA) CD25 positive selection kit according to the manufacturers’ protocols as previously described (18 (link)–20 (link)). One hundred thousand freshly isolated Tregs (in 200 microliters of normal saline) were administered to the mice 18 h prior to IRI (6 hours after injection of isotype or blocking antibodies).
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Modulating Immunity in Ischemic Injury
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Other organizations : University of Virginia Health System
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Variable analysis
independent variables
- Isotype control antibodies
- Blocking antibodies (anti-PD-L1, anti-PD-L2)
- Not explicitly mentioned
- Mice (WT)
- Timing of antibody administration (24 hours prior to IRI)
- Timing of Treg administration (18 hours prior to IRI, 6 hours after antibody injection)
- Treg isolation protocol (Dynal CD4 negative selection, Miltenyi Biotec CD25 positive selection)
- Positive control: Rat IgG2a (clone: 2A3) for anti-PD-L2
- Positive control: Rat IgG2b (clone: LTF-2) for anti-PD-L1
- Negative control: Untreated/saline-treated group not mentioned
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