Quantifying Human Monocyte Chemotaxis In Vitro and Murine Lymphocyte Trafficking In Vivo

• Published in: Journal of visualized experiments no. 128
• Main Authors: Prangley, Eliza, Kumar, Terrence, Ponda, Manish P
• Format: Journal Article
• Language: English
• Published: United States MyJove Corporation 30.10.2017
• Subjects: Animals; Chemotaxis, Leukocyte - physiology; Flow Cytometry - methods; Immunology; Lymphocytes - cytology; Lymphocytes - metabolism; Lymphocytes - physiology; Male; Mice; Mice, Inbred C57BL; Monocytes - cytology
• ISSN: 1940-087X; 1940-087X
• DOI: 10.3791/56218

Chemotaxis is migration along a specific chemical gradient . Chemokines are chemotactic cytokines that promote cellular trafficking with anatomic and temporal specificity . Chemotaxis is a critical function of lymphocytes and other immune cells that can be quantitatively assessed in vitro. This manuscript describes methods that permit the evaluation of chemotaxis, both in vitro and in vivo, for diverse cell types including cell lines and native cells. The in vitro, plate-based format permits the comparison of several conditions simultaneously in real-time, and can be completed within 1-4 h. In vitro assay conditions can be manipulated to introduce agonists and antagonists, as well as differentiate chemotaxis from chemokinesis, which is random movement. For in vivo trafficking assessments, immune cells can be labeled with multiple fluorescent dyes and used for adoptive transfer. The differential labeling of cells allows for mixed cell populations to be introduced into the same animal, thereby decreasing variance and reducing the number of animals required for an adequately powered experiment. Migration into lymphoid tissue occurs in as little as 1 h, and multiple tissue compartments can be sampled. Flow cytometry following tissue harvest allows for a rapid and quantitative analysis of the migratory patterns of multiple cell types.