Orthotopic Implantation and Peripheral Immune Cell Monitoring in the II-45 Syngeneic Rat Mesothelioma Model

• Published in: Journal of visualized experiments no. 104
• Main Authors: Weir, Chris J, Hudson, Amanda L, Peters, Lyndsay, Howell, Viive M
• Format: Journal Article
• Language: English
• Published: United States MyJove Corporation 02.10.2015
• Subjects: Animals; B-Lymphocytes - immunology; Cell Line, Tumor; Disease Models, Animal; Female; Flow Cytometry - methods; Killer Cells, Natural - immunology; Lung Neoplasms - immunology; Lung Neoplasms - pathology; Medicine; Mesothelioma - immunology; Mesothelioma - pathology; Mesothelioma, Malignant; Monitoring, Immunologic - methods; Monocytes - immunology; Neoplasm Transplantation - methods; Rats; Rats, Inbred F344; T-Lymphocyte Subsets - immunology
• ISSN: 1940-087X; 1940-087X
• DOI: 10.3791/53019

The enormous upsurge of interest in immune-based treatments for cancer such as vaccines and immune checkpoint inhibitors, and increased understanding of the role of the tumor microenvironment in treatment response, collectively point to the need for immune-competent orthotopic models for pre-clinical testing of these new therapies. This paper demonstrates how to establish an orthotopic immune-competent rat model of pleural malignant mesothelioma. Monitoring disease progression in orthotopic models is confounded by the internal location of the tumors. To longitudinally monitor disease progression and its effect on circulating immune cells in this and other rat models of cancer, a single tube flow cytometry assay requiring only 25 µl whole blood is described. This provides accurate quantification of seven immune parameters: total lymphocytes, monocytes and neutrophils, as well as the T-cell subsets CD4 and CD8, B-cells and Natural Killer cells. Different subsets of these parameters are useful in different circumstances and models, with the neutrophil to lymphocyte ratio having the greatest utility for monitoring disease progression in the mesothelioma model. Analyzing circulating immune cell levels using this single tube method may also assist in monitoring the response to immune-based treatments and understanding the underlying mechanisms leading to success or failure of treatment.