title: Studying the Role of Alveolar Macrophages in Breast Cancer Metastasis.
abstract: This paper describes the application of the syngeneic model of breast cancer (4T1) to the studies on a role of pulmonary alveolar macrophages in cancer metastasis. The 4T1 cells expressing GFP in combination with imaging and confocal microscopy are used to monitor tumor growth, track metastasizing tumor cells, and quantify the metastatic burden. These approaches are supplemented by digital histopathology that allows the automated and unbiased quantification of metastases. In this method the routinely prepared histological lung sections, which are stained with hematoxylin and eosin, are scanned and converted to the digital slides that are then analyzed by the self-trained pattern recognition software. In addition, we describe the flow cytometry approaches with the use of multiple cell surface markers to identify alveolar macrophages in the lungs. To determine impact of alveolar macrophages on metastases and antitumor immunity these cells are depleted with the clodronate-containing liposomes administrated intranasally to tumor-bearing mice. This approach leads to the specific and efficient depletion of this cell population as confirmed by flow cytometry. Tumor volumes and lung metastases are evaluated in mice depleted of alveolar macrophages, to determine the role of these cells in the metastatic progression of breast cancer.
authors.affiliation: Department of Immunotherapeutics and Biotechnology, School of Pharmacy, Texas Tech University Health Science Center.
authors.forename: Surya Kumari
authors.initials: SK
authors.lastname: Vadrevu
authors.affiliation: Department of Immunotherapeutics and Biotechnology, School of Pharmacy, Texas Tech University Health Science Center; Merck Research Labs.
authors.forename: Sharad
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Department of Immunotherapeutics and Biotechnology, School of Pharmacy, Texas Tech University Health Science Center; Department of Surgery, Memorial Sloan Kettering Cancer Center.
authors.forename: Navin
authors.initials: N
authors.lastname: Chintala
authors.affiliation: Department of Immunotherapeutics and Biotechnology, School of Pharmacy, Texas Tech University Health Science Center.
authors.forename: Jalpa
authors.initials: J
authors.lastname: Patel
authors.affiliation: Department of Immunotherapeutics and Biotechnology, School of Pharmacy, Texas Tech University Health Science Center; magdalena.karbowniczek@ttuhsc.edu.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Karbowniczek
authors.affiliation: Department of Immunotherapeutics and Biotechnology, School of Pharmacy, Texas Tech University Health Science Center; maciej.markiewski@ttuhsc.edu.
authors.forename: Maciej
authors.initials: M
authors.lastname: Markiewski
mesh_terms: D000818:Animals; D001943:Breast Neoplasms; D008081:Liposomes; D008168:Lung; D008175:Lung Neoplasms; D016676:Macrophages, Alveolar; D051379:Mice
chemical_list: D008081:Liposomes