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. 2022 Dec 16;8(50):eabn7983. doi: 10.1126/sciadv.abn7983

Fig. 2. Panitumumab treatment affects the TME in IBC tumors.

Fig. 2.

(A and B) Changes in CD8+ T cells, Tregs, and M2 macrophages in tissues of IgG2- and panitumumab-treated SUM149-hu-NSG-SGM3 mice analyzed by flow cytometry (A) and multiplexed immunofluorescence staining (B). (A) *P = 0.05 and **P < 0.001. (B) *P < 0.05. (C) Panitumumab-treated tissues have increased IFNG gene expression compared to IgG2-treated tissues in SUM149-hu-NSG-SGM3 mice. *P < 0.05. (D) Panitumumab-treated tissues have more CD3+CD8+G&B+ cells than IgG2-treated tissues in SUM149-hu-NSG-SGM3 mice as analyzed by multiplexed immunofluorescence staining. Scale bars, 50 μm. (E to G) Changes in CD8+ T cells, M2 macrophages, and Tregs after panitumumab treatment in matched tissues from three patients with IBC having a pCR (F) or five patients with IBC without a pCR (G) to panitumumab/NAC in primary HER2-negative IBC (NCT01036087). (E) Representative images of multiplexed immunofluorescence staining of CD3, CD8, CD68, CD163, FOXP3, and CK7 in IBC patient tissues at baseline (before panitumumab treatment, left) and week 2 (after panitumumab treatment, right). The numbers of CD8+ T cells (CD3+CD8+), M2 macrophages (CD68+CD163+), and Tregs (CD3+FOXP3+) in five randomly selected areas of each slide were calculated. Each symbol represents the same patient. Scale bars, 50 μm. (H) EGFR knockdown SUM149 clones shEGFR-4 and shEGFR-5 induce less migration of M2 macrophages and Tregs than control knockdown clone shCtrl in in vitro coculture transwell assays. *P < 0.01. G&B, granzyme B; DAPI, 4′,6-diamidino-2-phenylindole; FBS, fetal bovine serum. Experiments in (C) and (H) were independently repeated with three replicates each time.