Figure 2 from Estrogen Receptor Blockade Potentiates Immunotherapy for Liver Metastases by Altering the Liver Immunosuppressive Microenvironment

Estrogen depletion alters the cytokines/chemokines repertoire of the liver and increases expression of CCR5 and CCL5 in NKT and CD8⁺ T cells. A, Cytokine array analysis was conducted on whole liver protein lysates (four livers/group) derived from MC-38 tumor-bearing mice. Data were normalized to SHAM-operated values. In B–E are FC results for the indicated cell populations (n = 4). In F (left)—representative RNAscope images of frozen liver sections probed with CCL5 and NK1.1, followed by CD3 staining, and in F (right) image analysis results based on 15 fields per group (n = 3). Yellow arrows denote NK1.1⁺CCL5⁺CD3⁺ cells, and green arrows denote NK1.1⁺CD3⁺ cells with high CCL5 density. Experimental LM were generated by intrasplenic/portal injection of 10⁵ MC-38 cells. Intraperitoneal Mvc or vehicle injections began the following day and continued for 6 days. Mice were sacrificed and LM enumerated on day 18 post-inoculation. Results shown are of a representative experiment of two performed. The number of metastases per liver are shown in G (n = 4–7; bars indicate medians) and representative livers are shown in H. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001, Student t test (FC) and Mann–Whitney test (LM).