Flow Cytometry Detection of Chemokine Receptors for the Identification of Murine Monocyte and Neutrophil Subsets

Bonavita, Ornella; Massara, Matteo; Anselmo, Achille; Somma, Paolo; Brühl, Hilke; Mack, Matthias; Locati, Massimo; Bonecchi, Raffaella

doi:10.1016/bs.mie.2015.09.011

Cited by 1 publication

(2 citation statements)

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“…Blood was collected from the retro-orbital plexus and by cardiac puncture as described 53 . Briefly, blood was collected in 2KD-EDTA spray-coated tubes (BD Bioscience), washed in FACS buffer (PBS −/− , 1% bovine serum albumin, and 0.05 % sodium azide), red blood cells were lysed, washed again, and cells were stained as indicated.…”

Section: Methodsmentioning

confidence: 99%

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ACKR2 in hematopoietic precursors as a checkpoint of neutrophil release and anti-metastatic activity

et al. 2018

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Atypical chemokine receptors (ACKRs) are regulators of leukocyte traffic, inflammation, and immunity. ACKR2 is a scavenger for most inflammatory CC chemokines and is a negative regulator of inflammation. Here we report that ACKR2 is expressed in hematopoietic precursors and downregulated during myeloid differentiation. Genetic inactivation of ACKR2 results in increased levels of inflammatory chemokine receptors and release from the bone marrow of neutrophils with increased anti-metastatic activity. In a model of NeuT-driven primary mammary carcinogenesis ACKR2 deficiency is associated with increased primary tumor growth and protection against metastasis. ACKR2 deficiency results in neutrophil-mediated protection against metastasis in mice orthotopically transplanted with 4T1 mammary carcinoma and intravenously injected with B16F10 melanoma cell lines. Thus, ACKR2 is a key regulator (checkpoint) of mouse myeloid differentiation and function and its targeting unleashes the anti-metastatic activity of neutrophils in mice.

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Section: Methodsmentioning

confidence: 99%

“…Flow cytometry analysis was performed as previously described 53 . To exclude dead cells from analysis, cells were stained with Violet dead cell stain kit (Thermo Fisher).…”

Section: Methodsmentioning

confidence: 99%