Correction: Ponzo et al. Nucleolin Therapeutic Targeting Decreases Pancreatic Cancer Immunosuppression. Cancers 2022 , 14, 4265

Abstract

In the original publication [...].

Figure 3

N6L increases lymphocyte infiltration and activation in mPDAC tumours. (A) Tumour sections of control and N6L-treated mice were immunostained by an anti-CD8 antibody to detect CD8⁺ cells (arrows); scale bar: 50 μm. (B) CD8⁺ cells in tumoral regions (at least 4) of tumour slices were counted by using QuPath software, and the results were plotted as a mean for each tumour (n = 6) as the number of cells/mm². Two-tailed Mann-Whitney U-test (**, p < 0.01; n = 5 tumours) was applied. (C) Effector T cell activation was analysed by flow cytometry, and the % of CD3⁺IFNγ⁺ cells in control and N6L-treated mice is plotted (two-tailed Mann-Whitney U-test **, p < 0.01; n = 6 mice).

Figure 6

Anti-IL-6 antibody mimics N6L effects on PDAC immune microenvironment. mPDAC tumours were generated as in Figure 2 and treated with anti-IL-6 blocking antibody or IgG control antibody for three weeks, three times per week, by i.p. injection. Mice were sacrificed 21 days after the treatments. (A) Tumour volumes were measured as in Figure 2, and graphs show a representative experiment (from two independent experiments). (B–D) Immune cell populations in tumour tissues were analysed by flow cytometry as in Figure 2. Tregs and PMN-MDSC were analysed, and the results are plotted as the fold change of the % of (B) CD45⁺CD4⁺FoxP3⁺ cells and (C) CD11b⁺Ly6G⁺Ly6C^low cells in N6L-treated tumours relative to control tumours. (D) Fold change of the CD8/Treg ratio between control and N6L-treated tumours. (E) Tumour sections of control and anti-IL-6-treated mice were immunostained by an anti-CD8 antibody, CD8⁺ cells were counted (arrows), and results were plotted as (F) the number of cells/mm². Scale bar: 50 μm. p-values were calculated between indicated conditions by two-tailed Mann-Whitney t-test (**, p < 0.01; *, p < 0.05; n = 5 mice). (G) Effector T cell activation was analysed by flow cytometry as in Figure 3C, and the % of CD3⁺IFNγ⁺ cells in control and N6L-treated mice is plotted (two-tailed Mann-Whitney U-test **, p < 0.01; control, n = 5 mice; anti-IL-6, n = 4 mice).

Supplementary Figure S1

Characterization of mPDAC immune microenvironment. Immuno-competent syngenic FVB/n mice were injected with mPDAC cells into the pancreas. After three weeks, mice (n = 6) were sacrificed and tumours and draining lymph nodes were collected. (A) Tumour sections were immunostained an anti-CD45 antibody (arrows). (B,C) Immune cell populations of tumours and lymph nodes were analyzed by flow cytometry and frequency were calculated among CD45⁺ cells. Graphs show the % of CD45⁺CD8⁺, CD45⁺CD4⁺, CD45⁺CD11b⁺, CD45⁺CD4⁺FoxP3⁺ (Tregs among CD4⁺, and the ratio of CD8/Treg in tumours. All statistical tests are Two-tailed Mann-Whitney U-test; **, p < 0.01; ns = not significant, n = 6. Scale bars 50 μm.