Fig. 2. PlGF-2123–144 conjugation reduces systemic exposure to checkpoint blockade Abs and potential treatment-related toxicity. PlGF-2123–144 conjugation.

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Fig. 2. PlGF-2123–144 conjugation reduces systemic exposure to checkpoint blockade Abs and potential treatment-related toxicity. PlGF-2123–144 conjugation reduces systemic exposure to checkpoint blockade Abs and potential treatment-related toxicity. Mice were inoculated with 5 × 105 B16F10 cells on day 0. (A and B) PlGF-2123–144–αCTLA4 and PlGF-2123–144–αPD-L1 (100 μg each), αCTLA4 and αPD-L1 (100 μg each), or PBS was administered on day 4. PlGF-2123–144–Abs and PBS were injected peritumorally (pt), and unmodified Abs were injected either intraperitoneally (ip) or peritumorally. Blood plasma was collected on days 5, 7, 9, and 11. Concentrations of (A) αCTLA4 and (B) PD-L1 in blood plasma were determined by ELISA [PBS, n = 6; Abs (intraperitoneally), n = 5; others, n = 8; mean ± SEM]. (C to F) PlGF-2123–144–αCTLA4 or unmodified αCTLA4 and αPD-L1 (500 μg each per injection) were injected peritumorally on days 4 and 7. On day 9, blood serum was collected, and concentrations of (C) TNFα and (D) IFN-γ and (E) ALT activity in serum were measured (mean ± SEM). (F) Histologic liver sections were obtained on day 8. The number of lymphocytic infiltration spots was counted and divided by area (mean ± SEM). (G) Sixteen-week-old male NOD mice were given 100 μg of PlGF-2123–144–αPD-L1 or unmodified αPD-L1 on days 0 and 2 and evaluated for the development of diabetes. All Ab injections were intradermal at the back skin. Clinical diabetes was defined as blood glucose concentrations of 250 mg/dl for three consecutive days. Statistical analyses were done using ANOVA with Tukey’s test. Kruskal-Wallis test followed by Dunn’s multiple comparison was used in (C) due to nonparametric data. Log-rank (Mantel-Cox) test was performed for diabetes-free survival curves. Two experimental repeats. *P < 0.05, **P < 0.01. N.S., not significant. Jun Ishihara et al., Sci Transl Med 2017;9:eaan0401 Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works