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

Le Brocq, M. L., Fraser, A. R., Cotton, G., Woznica, K., McCulloch, C. V., Hewit, K. D., McKimmie, C. S., Nibbs, R. J.B. , Campbell, J. D.M. and Graham, G. J. (2014) Chemokines as novel and versatile reagents for flow cytometry and cell sorting. Journal of Immunology, 192(12), pp. 6120-6130. (doi: 10.4049/jimmunol.1303371) (PMID:24850722)

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Abstract

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.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Fraser, Dr Alasdair and Campbell, Dr John and Nibbs, Professor Rob and Le Brocq, Dr Michelle and Hewit, Dr Kay and McCulloch, Mrs Clare and McKimmie, Dr Clive and Graham, Professor Gerard
Authors: Le Brocq, M. L., Fraser, A. R., Cotton, G., Woznica, K., McCulloch, C. V., Hewit, K. D., McKimmie, C. S., Nibbs, R. J.B., Campbell, J. D.M., and Graham, G. J.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
College of Medical Veterinary and Life Sciences > School of Medicine, Dentistry & Nursing
Journal Name:Journal of Immunology
Publisher:American Association of Immunologists
ISSN:0022-1767
ISSN (Online):1550-6606

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
600961Dissecting the chemokine basis for the orchestration of the in vivo inflammatory responseGerard GrahamWellcome Trust (WELLCOME)099251/Z/12/ZIII -IMMUNOLOGY