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3. Results
3.1. CCL1 driven migration through CCR8 in activated eosinophils
To establish a model to monitor endogenous function of CCR8 functions, we evaluated primary eosinophils and an eosinophilic like cell line AML14.3D10. A subpopulation of fresh na?ve eosinophils expressed low surface levels of CCR8, while fresh AML14.3D10 Silmitasertib expressed almost undetectable levels of CCR8 (Fig. 1A). However, during differentiation towards an active eosinophilic phenotype the surface expression of CCR8 is markedly enhanced on AML14.3D10 cells (Fig. 1B). Maximum expression is reached at day 5 in culture when >60% of these cells express a high surface CCR8 level. Longer culture negatively influenced expression level (Fig. 1B). These data demonstrate for the first time that human eosinophils acquire CCR8 expression during maturation.
Fig. 1.
Expression of surface CCR8 on na?ve human eosinophils and undifferentiated AML cells (A) and differentiated AML cells (B) was evaluated by flow cytometry. Percentage CCR8 positive cells as compared to isotype staining is shown. Chemotaxis of na?ve human eosinophils (C) and fully differentiated AML cells (D) in response to CCL1 and CCL11. Values in C and D indicate the maximum specific response to the given chemokine. Data show one representative experiment of at least three. Standards variation was routinely less than 20%.