TGN1412 Induces Lymphopenia and Human Cytokine Release in a Humanized Mouse Model
Fig 5
Humanized mice succumb to antibody application.
Humanized mice were injected i.v. with 20 μg OKT3 or 20 μg TGN1412 per 10 grams body weight. 1 hour before, indicated humanized mice were treated with 10 μg anti-human TNF-α mAb per mouse (C-E). As control, humanized mice were injected with PBS (A-B). (A) Before and 2–6 hours (time point of sacrifice) post OKT3 (n = 6) or TGN1412 (n = 6) application, body temperature of mice was monitored. PBS-treated mice (n = 5) were used as control. ** p < 0.01; *** p < 0.001 (Dunnett-adjusted t-test). (B) 2–6 hours post OKT3 (n = 22) or TGN1412 (n = 22) application mice were sacrificed if severe signs of illness were detectable. PBS-treated mice (n = 18) were sacrificed 6 hours post treatment, the latest time point survived by mAb-treated mice. *** p < 0.001 (Dunnett-HSU-adjusted Logrank test). (C) Before (be.) and after (af.) OKT3 (n = 11) or TGN1412 (n = 6) application and 1 hour post anti-human TNF-α mAb (α-hTNF-α) application (before OKT3 or TGN1412 treatment) and 0.33–7 hours (time point of sacrifice) post OKT3 (n = 14) or TGN1412 (n = 11) application, body temperature of mice was monitored. * p < 0.05; *** p < 0.001 (paired t-test for difference after-before). (D) 0.33–7 hours post OKT3 (n = 11), anti-human TNF- α mAb + OKT3 (n = 14), TGN1412 (n = 6), or anti-human TNF-α mAb + TGN1412 (n = 11) application mice were sacrificed if severe signs of illness were detectable. (E) Before (be.) and after (af.) OKT3 (n = 11) or TGN1412 (n = 6) application and 1 hour post anti-human TNF-α mAb application (before OKT3 or TGN1412 treatment; black bars) and 0.33–7 hours post OKT3 (n = 14) or TGN1412 (n = 11) application (time point of sacrifice; gray bars), percentages of hCD45+ cells in peripheral blood of reconstituted mice were analyzed by flow cytometric analysis. * p < 0.05; ** p < 0.01; *** p < 0.001 (Wilcoxon signed rank test for difference after-before). Data shown in (A) are taken from 2, in (B) from 6–9, and in (C), (D), and (E) from 3 independent experiments.