Comprehensive assessment of chemokine expression profiles by flow cytometry.
The chemokines are a large family of mainly secreted molecules involved in the regulation of numerous physiological and pathophysiological processes. Despite many years of investigation, the precise cellular sources of most chemokines have remained incompletely defined as a consequence of the limited availability of suitable reagents to visualize the expression of chemokine proteins at the single-cell level. Here, we developed a simple flow cytometry-based assay using commercially available chemokine-specific antibodies for efficient cell-associated detection of 37 of 39 murine chemokines. To demonstrate the utility of this methodology, we used it to reevaluate the nature of homeostatic chemokines in the hematopoietic compartment, to delineate the complete chemokine profiles of NK cells and B cells in response to major polyclonal stimuli, and to assess the chemokine response of DCs to bacterial infection. The versatility of this analytical methodology was further demonstrated by its application to selected human chemokines and should greatly facilitate any future investigation into chemokine biology at large.
Eberlein
Jens
J
Barbara Davis Center for Childhood Diabetes, Department of Medicine, University of Colorado Denver, Aurora, 80045-0511, USA.
Nguyen
Tom T
TT
Victorino
Francisco
F
Golden-Mason
Lucy
L
Rosen
Hugo R
HR
Homann
Dirk
D
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D019556:COS Cells; D002460:Cell Line; D018925:Chemokines / Q000096:biosynthesis* / Q000276:immunology; D002522:Chlorocebus aethiops; D005434:Flow Cytometry / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D007694:Killer Cells, Natural / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout
D000906:Antibodies; D018925:Chemokines