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OMIP-018

Chemokine receptor expression on human T helper cells

Tess Brodie, Elena Brenna, Federica Sallusto Cytometry PART A, Volume 83A, Issue 6, 530-532 (2013)

PURPOSE: This panel was developed for the enumeration of chemokine receptor expression on CD4 T cells from the naïve (TN), stem cell memory (TSCM), central memory (TCM), and effector memory (TEM) cell subsets (Table 1). Eight chemokine receptors were chosen based upon previously published observations implicating their preferential expression on T helper type 1 (Th1), Th2, Th17, or Th22 cells, and their role in controlling lymphocyte migration to secondary lymphoid tissues, B cell follicles or non-lymphoid tissues, both in the steady state and during immune responses to pathogens, autoantigens, or allergens (1). This panel has been optimized for use with fresh peripheral blood mononuclear cells (PBMCs) with the observation that 24 h after blood draw, the expression of some chemokine receptors is reduced. Optimal staining requires PBMCs to be processed and acquired within 5 h of blood draw and may be performed in a 96-well plate format for high throughput experiments.

CELL TYPE: Fresh PBMC

MACHINE: BD LSR Fortessa

Marker Target Clone Fluorochrome Purpose
ccr7 Human G043H7 Brilliant Violet 421 maturity
CD194 Human 1G1 PE-Cy7 polarization
CD183 Human 1C6/CXCR3 PE-Cy5 polarization
CCR5 Human 2D7/CCR5 APC-Cy7 polarization
CD193 Human 61828 APC polarization
CD294 Human BM16 FITC polarization
CD3 Human PE-Cy5.5 T cell
CD4 Human S3.5 PE-Texas Red T cell
CD45RA Human MEM-56 Qdot 655 maturity
CD95 Human Dx2 PerCP-eFluor 710 maturity
CCR6 Human 11A9 Biotin polarization
Biotin Human Qdot 800 secondary reagent to CCR6
CCR10 Human 314305 PE polarization
CD14 Human M5E2 V500 exclusion
CD16 Human 3G8 V500 exclusion
CD19 Human HIB19 V500 exclusion
Dead cells Human Live/Dead Fix Aqua viability/exclusion
CD185 Human 51505 Qdot 605 polarization