Figure 2.
rLIF administration decreases tissue inflammation and donor immune cell infiltration after allo-BMT. Lethally irradiated B6C3F1 and C57BL/6 mice received BM and BM + T cells from C57BL/6 and BALB/c mice, respectively, along with or without rLIF treatment. (A-B) BM+T increased spleen weight (A) and length of SI (B) in B6C3F1 mice and C57BL/6 mice at 7 days and 10 days after allo-BMT, respectively, which was largely abolished by rLIF administration. (A, left) Representative images of spleen tissues. For B6C3F1 mice: n = 4 for BM; n = 16 for both BM + T and BM + T + rLIF; for C57BL/6 mice: n ≥ 4 for BM; n ≥ 5 for both BM + T and BM + T + rLIF. (C) rLIF administration reduced the expression of majority inflammatory cytokines examined in B6C3F1 mice at 7 days after allo-BMT. Relative mRNA expression levels of TNFα, CXCL1, CCL2, IL-1a, IL-1b, and IL-22 in SI were determined by quantitative real-time PCR assays and normalized with β-actin. n ≥ 5 mice/group. (D) rLIF administration decreased the infiltration of donor CD45+ immune cells in epithelium of the intestine (EPI) from B6C3F1 mice at 7 days after allo-BMT. Representative flow cytometry images (left) and quantifications (right) show the percentage of donor CD45+ immune cells (H2kk–CD45+) in EPI from B6C3F1 mice at 7 days after allo-BMT. n = 8 mice/group. (E-F) rLIF administration decreased the donor immune cell infiltration in B6C3F1 (left) and C57BL/6 (right) mice at 7 and 10 days after allo-BMT, respectively. (E) The numbers of infiltrating donor cells in spleen, MLN, and LP tissues determined by flow cytometric assays. (F) The numbers of a set of infiltrating donor immune cells, including CD45, CD4, CD8, neutrophil, DC, macrophage and natural killer (NK) cells, in spleen (upper), MLN (middle), and LP (lower) tissues. The number of cells in mice that received BM + T without rLIF treatment was defined as 1. Gating strategies are shown in supplemental Figure 3. n ≥ 7 mice/group. Data are presented as mean ± standard deviation from at least 3 independent experiments. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, unpaired t test with Welch’s correction.

rLIF administration decreases tissue inflammation and donor immune cell infiltration after allo-BMT. Lethally irradiated B6C3F1 and C57BL/6 mice received BM and BM + T cells from C57BL/6 and BALB/c mice, respectively, along with or without rLIF treatment. (A-B) BM+T increased spleen weight (A) and length of SI (B) in B6C3F1 mice and C57BL/6 mice at 7 days and 10 days after allo-BMT, respectively, which was largely abolished by rLIF administration. (A, left) Representative images of spleen tissues. For B6C3F1 mice: n = 4 for BM; n = 16 for both BM + T and BM + T + rLIF; for C57BL/6 mice: n ≥ 4 for BM; n ≥ 5 for both BM + T and BM + T + rLIF. (C) rLIF administration reduced the expression of majority inflammatory cytokines examined in B6C3F1 mice at 7 days after allo-BMT. Relative mRNA expression levels of TNFα, CXCL1, CCL2, IL-1a, IL-1b, and IL-22 in SI were determined by quantitative real-time PCR assays and normalized with β-actin. n ≥ 5 mice/group. (D) rLIF administration decreased the infiltration of donor CD45+ immune cells in epithelium of the intestine (EPI) from B6C3F1 mice at 7 days after allo-BMT. Representative flow cytometry images (left) and quantifications (right) show the percentage of donor CD45+ immune cells (H2kk–CD45+) in EPI from B6C3F1 mice at 7 days after allo-BMT. n = 8 mice/group. (E-F) rLIF administration decreased the donor immune cell infiltration in B6C3F1 (left) and C57BL/6 (right) mice at 7 and 10 days after allo-BMT, respectively. (E) The numbers of infiltrating donor cells in spleen, MLN, and LP tissues determined by flow cytometric assays. (F) The numbers of a set of infiltrating donor immune cells, including CD45, CD4, CD8, neutrophil, DC, macrophage and natural killer (NK) cells, in spleen (upper), MLN (middle), and LP (lower) tissues. The number of cells in mice that received BM + T without rLIF treatment was defined as 1. Gating strategies are shown in supplemental Figure 3. n ≥ 7 mice/group. Data are presented as mean ± standard deviation from at least 3 independent experiments. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, unpaired t test with Welch’s correction.

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