European Urology Oncology

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European Urology Oncology Mannose Receptor–positive Macrophage Infiltration Correlates with Prostate Cancer Onset and Metastatic Castration-resistant Disease Jelani C. Zarif, Javier A. Baena-Del Valle, Jessica L. Hicks, Christopher M. Heaphy, Igor Vidal, Jacob Luo, Tamara L. Lotan, Jody E. Hooper, William B. Isaacs, Kenneth J. Pienta, Angelo M. De Marzo European Urology Oncology DOI: 10.1016/j.euo.2018.09.014 Copyright © 2018 Terms and Conditions

Fig. 1 Analytical validation of immunohistochemistry detection of CD206 expression. (A) Human M2 macrophages were stained for CD206 as a positive control and (B) PC3 cells were stained as a negative control. (C–H) Normal human prostate tissue and prostate cancer tissue were assessed for CD206 positivity. (C) Normal prostate tissue. (D) In benign regions with somewhat higher levels of inflammatory infiltrates (predominantly mononuclear) there was an increase in CD206-positive cells. (E) Normal prostate. (F) Gleason 3+4+7 core. (G) Lymph node metastasis. (H) Distant metastasis. European Urology Oncology DOI: (10.1016/j.euo.2018.09.014) Copyright © 2018 Terms and Conditions

Fig. 2 Immunofluorescent staining for CD206-positive macrophages. Deparaffinized prostate sections were stained for three markers. (A) Infiltration of cells with dual CD206/CD68 positivity cells (arrows) within the prostatic stromal compartment and the lumen (Lu); CD206-negative cells (short arrow) were also present. (B) D240 was used as a marker for the lumen of lymphatic vessels. Cells with dual CD206/D240 positivity (arrows) surround the lymphatic lumen; CD206-positive/D240-negative cells (short arrows) were present but not fused to the lymphatic vessel wall. 4′,6-Diamidino-2-phenylindole (DAPI) staining was also used for nuclei in (A) and (B). European Urology Oncology DOI: (10.1016/j.euo.2018.09.014) Copyright © 2018 Terms and Conditions

Fig. 3 Mannose receptor expression was greater in prostate cancer metastasis relative to normal benign prostate tissue. Using prostate cancer tissue microarrays and immunohistochemistry, MRC1/CD206 expression was quantified and compared for normal human prostate tissue and prostate cancer tissue. Comparisons were performed using a Mann-Whitney test and/or t test as appropriate. Mets=metastases. European Urology Oncology DOI: (10.1016/j.euo.2018.09.014) Copyright © 2018 Terms and Conditions

Fig. 4 Dual mannose receptor and scavenger receptor expression in metastatic castration-resistant prostate cancer (mCRPC) rapid autopsy samples obtained 0–4h after death. M2 macrophage infiltration was determined via flow cytometry. Coronal lymph node in mCRPC with (A) CD163 staining and (B) CD206 staining. Paratracheal lymph node in mCRPC with (C) CD163 staining and (D) CD206 staining. First right rib in mCRPC with (E) CD163 staining, (F) CD206 staining, and (G) dual CD206 and CD163 staining. European Urology Oncology DOI: (10.1016/j.euo.2018.09.014) Copyright © 2018 Terms and Conditions