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. 2024 Feb 21;132(2):027007. doi: 10.1289/EHP12758

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Figure 3A is a timeline depicting the protocol of oral tolerance induction of ovalbumin followed by a de novo challenge to ovalbumin in C 57 B L slash 6 J mice daily for 60 days exposed to the vehicle or to food-grade silicon dioxide at 1, 10, or 100 milligrams per kilogram of body weight per day through gavage. Figures 3B to 3J are box plots, plotting optical density 450 nanometers, ranging from 0.0 to 1.0 in increments of 0.5; lipocalin-2 picogram per milligram of feces, ranging from 0 to 20,000 in increments of 5,000; interleukin-1 lowercase beta relative expression, ranging from 0 to 20 in increments of 5; interferon-gamma picograms per milliliter, ranging from 0 to 25 in increments of 5; interleukin-10 relative expression, ranging from 0 to 3 in unit increments; transforming growth factor lowercase beta relative expression, ranging from 0.0 to 2.5 in increments of 0.5; interleukin-10 picograms per milliliter, ranging from 0 to 10 in increments of 2; transforming growth factor lowercase beta picograms per milliliter, ranging from 0 to 400 in increments of 100; and transforming growth factor lowercase beta picograms per milligram of tissue, ranging from 0 to 10 in increments of 2 (y-axis) across food-grade silicon dioxide (milligrams per kilogram), including P B S at 0 and 0 to 1 in unit increments, 1 to 10 in increments of 9, 10 to 100 in increments of 90 for O V A (x-axis) for immunoglobulin G-anti O V A. — Evaluation of oral tolerance and gut immune response to ovalbumin in mice exposed 60 d to food-grade ${SiO}_{2}$ . (A) The protocol of oral tolerance induction to ovalbumin (OVA) followed by a de novo challenge to OVA in C57BL/6J mice daily exposed to the vehicle (water) or to the food-grade ${SiO}_{2}$ ( $food-grade silicon dioxide$ ) (1, 10, or $100 milligrams per kilogram$ BW/d) through gavage. (B) OVA-specific IgG secretion in OVA-immunized (PBS) or OVA-tolerized mice exposed to a vehicle or $food-grade silicon dioxide$ for 60 d ( $lowercase italic n equals 7$ mice per group). (C) Fecal lipocalin-2 (Lcn2) levels in OVA-immunized (PBS) or OVA-tolerized mice exposed to a vehicle or $food-grade silicon dioxide$ for 60 d ( $lowercase italic n equals 7$ mice per group). (D) $interleukin 1 lowercase beta$ transcript expression in colon of OVA-immunized (PBS) or OVA-tolerized mice exposed to a vehicle or $food-grade silicon dioxide$ for 60 d ( $lowercase italic n equals 7$ mice per group). (E) Amounts of $IFN- γ$ secreted by mesenteric lymph node (MLN) cells from OVA-immunized (PBS) or OVA-tolerized mice exposed to a vehicle or $food-grade silicon dioxide$ for 60 d ( $lowercase italic n equals 7$ mice per group). (F,G) Il10 (F) and $T g f β$ (G) transcript expression in colon of OVA-immunized (PBS) or OVA-tolerized mice exposed to a vehicle or $food-grade silicon dioxide$ for 60 d ( $lowercase italic n equals 7$ mice per group). (H,I) Amounts of IL-10 (H) and $TGF- β$ (I) secreted by mesenteric lymph node cells from OVA-immunized (PBS) or OVA-tolerized mice exposed to a vehicle or $food-grade silicon dioxide$ for 60 d ( $lowercase italic n equals 7$ mice per group). Secreted $TGF- β$ amounts by colon explants from OVA-immunized (PBS) or OVA-tolerized mice exposed to a vehicle or $food-grade silicon dioxide$ for 60 d ( $lowercase italic n equals 7$ mice per group). The data are expressed as median with interquartile range and whiskers extending from minimum to maximum ± SEM. * $lowercase italic p less than 0.05$ ; ** $lowercase italic p less than 0.01$ ; *** $lowercase italic p less than 0.001$ by one-way ANOVA and post hoc Tukey test (E–J) or Kruskal-Wallis test followed by a post hoc Dunn’s test (C,D). Data behind graphs are reported in Table S4. Note: ANOVA, analysis of variance; BW, body weight; CFA, complete Freund adjuvant; IgG, immunoglobulin G; $IFN- γ$ , interferon gamma; IL-10, interleukin 10; OD, optical density; PBS, phosphate-buffered saline; SEM, standard error of the mean; $TGF- β$ , transforming growth factor beta.