Circulating CD14+CD204+ Cells Predict Postoperative Recurrence in Non–Small-Cell Lung Cancer Patients  Ryo Maeda, MD, Genichiro Ishii, MD, PhD, Shinya.

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Circulating CD14+CD204+ Cells Predict Postoperative Recurrence in Non–Small-Cell Lung Cancer Patients  Ryo Maeda, MD, Genichiro Ishii, MD, PhD, Shinya Neri, MD, Kazuhiko Aoyagi, PhD, Hironori Haga, MD, PhD, Hiroki Sasaki, MD, PhD, Kanji Nagai, MD, PhD, Atsushi Ochiai, MD, PhD  Journal of Thoracic Oncology  Volume 9, Issue 2, Pages 179-188 (February 2014) DOI: 10.1097/JTO.0000000000000044 Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 1 Cell surface protein expression of CD204 on CD14+ cells from PA cultured in lung cancer cell line–conditioned medium. A, Blood samples from ligated PA of surgically resected lungs. Change in cell surface protein expressions of CD204 on CD14+ cells as determined using flow cytometry. B, Blood samples from ligated PA of surgically resected lungs. Change in cell surface protein expressions of CD204 on CD14+ cells as determined using flow cytometry. B, Change in CD204 cells per CD14 cell ratio (n = 4 for each group). *Compared with cases cultured in fibroblast-conditioned medium (all, p < 0.05). PA, pulmonary artery; FITC, fluorescein isothiocyanate. Journal of Thoracic Oncology 2014 9, 179-188DOI: (10.1097/JTO.0000000000000044) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 2 Correlation among the number of CD204+ macrophages infiltrating the stroma of the primary tumor and the recurrence-free probability in stage I adenocarcinoma patients. A, Immunohistochemical staining for CD204 in lung cancer specimens. The sections were obtained from tumor specimens with low or high levels of CD204+ cells. B, Significantly more cases with a high number of CD204+ macrophages develop early recurrence within 3 years after resection (p < 0.01).C, CD204+ macrophages are found not only in the stroma of the tumor but also in intratumoral blood vessels. H.E., hematoxylin and eosin staining (×200); VVG, Victoria blue van Gieson staining (×200); CD204, CD204 staining (×200). Journal of Thoracic Oncology 2014 9, 179-188DOI: (10.1097/JTO.0000000000000044) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 3 Correlation among the number of CD204+ macrophages infiltrating the stroma of the primary tumor, the number of CD14+CD204+ cells from the PV, and the probability of postoperative recurrence in lung cancer patients. A, Quantification of CD14+CD204+ cells among blood mononuclear cells from the PV. Representative data for two flow cytometry analyses. B, The number of CD14+CD204+ cells from the PV is correlated with the number of CD204+ macrophages in the stroma of the primary tumor. R = 0.658. *p < 0.05. C, A significantly greater number of CD204+ macrophages in the stroma are observed among the patients with high levels of CD14+CD204+ cells from the PV. D, Significantly more cases with a high number of CD14+CD204+ cells from the PV develop early recurrence within 2 years after resection (p < 0.01). PV, pulmonary vein. Journal of Thoracic Oncology 2014 9, 179-188DOI: (10.1097/JTO.0000000000000044) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions

FIGURE 4 CD14+CD204+ cells facilitated the lung metastasis of cancer cells more effectively than CD14+CD204− cells. A, Matrigel invasion assay. A significantly increased number of invading A549 cells on the lower surface of the filters is identified when CD14+CD204+ cells are added. *p < 0.05. B, CD14+CD204+ cells promote the metastasis of A549 cancer cells. *p < 0.05. C, Our concept that CD204+ tumor–associated macrophages remote from the primary tumor directly contribute to the metastasis of cancer cells at distant sites. Journal of Thoracic Oncology 2014 9, 179-188DOI: (10.1097/JTO.0000000000000044) Copyright © 2014 International Association for the Study of Lung Cancer Terms and Conditions