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. 2022 Oct 4;13:1008390. doi: 10.3389/fimmu.2022.1008390

Figure 2.

Figure 2

In vivo analysis in LPS-transient ALI rhesus macaque model studies. (A) Diagram of in vivo study along timeline, t-0 (baseline) to 72hrs with measures obtained. LPS [50 μg/kg] was infused intravenously (IV) after a IV single dose of humanized anti-DEspR (hu6g8) antibody (n = 3), or mock-treatment (Tx) saline for placebo LPS-control (n = 2). Non-LPS PBS-only (n = 1) served as negative control. Measures obtained at designated time points: plasma TNF-α, IL-6; body temperature (temp), hypoxemia (% O2-saturation), flow cytometry analysis of % DEspR+CD11b+ neutrophils (Ns) in BALF, bronchoalveolar lavage fluid and whole blood samples, and absolute total number (#) of peripheral DEspR+CD11b+ neutrophils (K/μL) in whole blood samples. BL, baseline levels, NL, normal levels; 0, zero levels; ↓, decrease; + to +++, increased levels with higher +++; h, hrs; Δ*, significant change between treated and saline mock-treated macaques in corresponding parameter. (B) Pharmacokinetic analysis of anti-DEspR hu6g8 in macaques (n = 2) given 3 mg/kg dose IV. Half-life of distribution, t ½ α: 18.6 hours (h), half-life of elimination, t ½ β: 12.1 days (D). Time points for human IgG4 ELISA measurements in another study group of macaques (n = 2): 2-, 4-, 8-, 24-, 72-, 144-, 312-, 480-hours. Time course of averages of different measures per study group plotted (C-N), n = 3 LPS + treatment (LPS-Tx), n = 2 LPS-saline control, n = 1 no-LPS, no treatment control. One-way repeated measures ANOVA, with Holm-Sidak pairwise comparisons performed, significant P values notated respectively. (C) ELISA plasma TNF-α (ng/ml); (D) ELISA plasma interleukin (IL)-6 (ng/ml); (E) body temperature (°C); (F) O2 saturation (SpO2): *, P = 0.03. (G) neutrophil-lymphocyte ratio (NLR), (H) peripheral % DEspR+CD11b+ neutrophils (Ns): 24h (*) P = 0.02; 72h (**) P = 0.03); (I) peripheral total number (#) of DEspR+CD11b+ Ns (K/μL): 8h (*) P = 0.03; 24h (***) P = 0.0001; 72h (*) P = 0.01. (J) BALF % DEspR+CD11b+ Ns: 4h: (*) P = 0.02. (K) Peripheral absolute neutrophil counts (ANC), (L) % DEspR-negative [-] neutrophils (Ns): 24h (**), P = 0.008, and 72h (**) P = 0.007. (M) Peripheral number (#) of DEspR[-] Ns, and (N) BALF % DEspR[-] Ns: 4h (*) P = 0.02.