Fig. 3

ICI-induced irAEs are amplified upon non-specific CTLA4 blockade. Adverse events were examined in unilaterally inoculated mice administered 5 mg/kg anti-CTLA4 into the tbLN or the ntbLN using a LDDS or through i.p. on day 4 and 8 post tumor inoculation. A. Body weight changes. Body weight was normalized to the day 0 value for each mouse. (Control, n = 14; tbLN, n = 10; ntbLN, n = 11; i.p., n = 8) B. Spleen index. Spleen index was obtained by dividing the weight of spleen by the body weight. (No tumor control, n = 12; Control, n = 6; tbLN, n = 10; ntbLN, n = 7; i.p., n = 4) C. Level of IL6 (Control, n = 5; tbLN, n = 6; ntbLN, n = 6; i.p., n = 7) and D. IFNγ in the spleen (Control, n = 5; tbLN, n = 7; ntbLN, n = 6; i.p., n = 5). E. Severity of different irAEs as confirmed by examination of stained tissue section. (Pneumonia: No tumor control, n = 17; Control, n = 20; tbLN, n = 11; ntbLN, n = 11; i.p., n = 5; Glomerulonephritis and Vasculitis: No tumor control, n = 17; Control, n = 13; tbLN, n = 11; ntbLN, n = 11; i.p., n = 5; Sialadenitis: No tumor control, n = 17; Control, n = 13; tbLN, n = 9; ntbLN, n = 11; i.p., n = 5) F. HE (Hematoxylin and Eosin)- or EM (Elastica Masson)-stained tissue specimen showing frequently observed irAEs in the case of anti-CTLA4 administered using different strategies. Statistical analyses were done using ANOVA with Tukey’s test. *P < 0.05, **P < 0.01, and ****P < 0.0001. Data are represented as means ± SEM