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. 2023 Mar 17;42(3):112269. doi: 10.1016/j.celrep.2023.112269

Figure 4.

Figure 4

ILC3s are required to prevent intestinal bacterial spread to liver and spleen

(A and B) (A) SPF Rorc(γt)−/+ and Rorc(γt)−/− littermate mice and (B) SPF Rag1−/− or Rorc(γt)−/−Rag2−/− mice underwent PH. Antibiotic-resistant E. coli JM83 was intragastrically (i.g.) delivered 12 h before PH for efficient trackability. E. coli titers (CFU) were assessed in the liver and spleen 24 h after PH.

(C) Experimental setup used in experiments for Thy 1.2 cell depletion using anti-Thy 1.2 or anti-immunoglobulin G (IgG) antibody administered 6 days, 3 days and 1 day prior to PH in SPF (D and E) wild-type or (F–H) Rag1−/− mice. Twelve hours before the PH or sham surgery, 1010 CFU of E. coli JM83 was i.g. delivered to the mice.

(D) Depletion efficiency of ILC3s in wild-type mice at day 0 (d0).

(E) E. coli titers (CFU) in the liver following Thy1.2 depletion in wild-type mice 24 h after PH.

(F) Depletion efficiency of ILC3s in Rag1−/− at d0.

(G and H) E. coli titers (CFU) in (G) MLN and (H) liver and spleen following Thy1.2 depletion in Rag1−/− mice 24 h after PH.

(I–K) Sort-purified CD19+ B cells and CD3+CD5+ T cells from spleen and MLN of Thy1.1 mice were transferred i.v. to Rag1−/− Thy1.2 recipient mice, and i.p. injection of anti-Thy1.2 was performed three times prior to PH. (I and J) Depletion efficiency of Thy 1.2+ cells and presence of T and B cells at d0. (K) E. coli titers (CFU) in the liver and spleen 24 h after PH.

(L) Expression of IL1b and IL18 in sham-operated or PH wild-type mice determined by RT-qPCR.

(M) Colonized wild-type, IL-18−/−, and Asc−/− underwent PH. Antibiotic-resistant E. coli JM83 was i.g. delivered 12 h before PH. E. coli titers (CFU) were assessed in the liver and spleen 24 h after PH. Geometric means for log scales and arithmetic means for linear scales are shown. Normalized values were analyzed by Student’s t test to compare two experimental groups or by ANOVA to compare more than two groups in parallel. Data in (D)–(K) are representative of n = 3–5 mice per group in two independent experiments, and data in (A), (B), and (M) are pooled based on n = 5–17 mice from two or more independent experiments. The p values are indicated as follows: ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.001, and ∗∗∗∗p ≤ 0.0001. See also Figure S4.