Chemokines as novel and versatile reagents for flow cytometry and cell sorting

• Published in: The Journal of immunology (1950) Vol. 192; no. 12; pp. 6120 - 6130
• Main Authors: Le Brocq, Michelle L, Fraser, Alasdair R, Cotton, Graham, Woznica, Kerry, McCulloch, Clare V, Hewitt, Kay D, McKimmie, Clive S, Nibbs, Robert J B, Campbell, John D M, Graham, Gerard J
• Format: Journal Article
• Language: English
• Published: United States 15.06.2014
• Subjects: Animals; Chemokine CCL19 - chemistry; Chemokine CCL19 - immunology; Female; Flow Cytometry - methods; Humans; Male; Mice; Receptors, CCR7 - chemistry; Receptors, CCR7 - immunology
• DOI: 10.4049/jimmunol.1303371
• ISSN: 0022-1767

Cell therapy regimens are frequently compromised by low-efficiency cell homing to therapeutic niches. Improvements in this regard would enhance effectiveness of clinically applicable cell therapy. The major regulators of tissue-specific cellular migration are chemokines, and therefore selection of therapeutic cellular populations for appropriate chemokine receptor expression would enhance tissue-homing competence. A number of practical considerations preclude the use of Abs in this context, and alternative approaches are required. In this study, we demonstrate that appropriately labeled chemokines are at least as effective in detecting their cognate receptors as commercially available Abs. We also demonstrate the utility of biotinylated chemokines as cell-sorting reagents. Specifically, we demonstrate, in the context of CCR7 (essential for lymph node homing of leukocytes), the ability of biotinylated CCL19 with magnetic bead sorting to enrich for CCR7-expressing cells. The sorted cells demonstrate improved CCR7 responsiveness and lymph node-homing capability, and the sorting is effective for both T cells and dendritic cells. Importantly, the ability of chemokines to detect CCR7, and sort for CCR7 positivity, crosses species being effective on murine and human cells. This novel approach to cell sorting is therefore inexpensive, versatile, and applicable to numerous cell therapy contexts. We propose that this represents a significant technological advance with important therapeutic implications.