Abstract
Isolation of murine neutrophils from several anatomical compartments allows for functional characterization and analysis of these cells. Here we describe the isolation of bone marrow, peripheral blood, and lung airspace and interstitial neutrophil populations, using density gradient separation, lavage, and flow cytometry techniques.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Amulic B, Cazalet C, Hayes GL, Metzler KD, Zychlinsky A (2012) Neutrophil function: from mechanisms to disease. Annu Rev Immunol 30:459–489. https://doi.org/10.1146/annurev-immunol-020711-074942
Ley K, Laudanna C, Cybulsky MI, Nourshargh S (2007) Getting to the site of inflammation: the leukocyte adhesion cascade updated. Nat Rev Immunol 7(9):678–689. https://doi.org/10.1038/nri2156
Mantovani A, Cassatella MA, Costantini C, Jaillon S (2011) Neutrophils in the activation and regulation of innate and adaptive immunity. Nat Rev Immunol 11(8):519–531. https://doi.org/10.1038/nri3024
Summers C, Rankin SM, Condliffe AM, Singh N, Peters AM, Chilvers ER (2010) Neutrophil kinetics in health and disease. Trends Immunol 31(8):318–324. https://doi.org/10.1016/j.it.2010.05.006
Suratt BT, Young SK, Lieber J, Nick JA, Henson PM, Worthen GS (2001) Neutrophil maturation and activation determine anatomic site of clearance from circulation. Am J Physiol Lung Cell Mol Physiol 281(4):L913–L921
Kordonowy LL, Burg E, Lenox CC, Gauthier LM, Petty JM, Antkowiak M, Palvinskaya T, Ubags N, Rincon M, Dixon AE, Vernooy JH, Fessler MB, Poynter ME, Suratt BT (2012) Obesity is associated with neutrophil dysfunction and attenuation of murine acute lung injury. Am J Respir Cell Mol Biol 47(1):120–127. https://doi.org/10.1165/rcmb.2011-0334OC
Palvinskaya T, Antkowiak M, Burg E, Lenox CC, Ubags N, Cramer A, Rincon M, Dixon AE, Fessler MB, Poynter ME, Suratt BT (2013) Effects of acute and chronic low density lipoprotein exposure on neutrophil function. Pulm Pharmacol Ther 26(4):405–411. https://doi.org/10.1016/j.pupt.2012.10.002
Ubags ND, Burg E, Antkowiak M, Wallace AM, Dilli E, Bement J, Wargo MJ, Poynter ME, Wouters EF, Suratt BT (2016) A comparative study of lung host Defense in murine obesity models. Insights into neutrophil function. Am J Respir Cell Mol Biol 55(2):188–200. https://doi.org/10.1165/rcmb.2016-0042OC
Zaynagetdinov R, Sherrill TP, Kendall PL, Segal BH, Weller KP, Tighe RM, Blackwell TS (2013) Identification of myeloid cell subsets in murine lungs using flow cytometry. Am J Respir Cell Mol Biol 49(2):180–189. https://doi.org/10.1165/rcmb.2012-0366MA
Abdulnour RE, Sham HP, Douda DN, Colas RA, Dalli J, Bai Y, Ai X, Serhan CN, Levy BD (2016) Aspirin-triggered resolvin D1 is produced during self-resolving gram-negative bacterial pneumonia and regulates host immune responses for the resolution of lung inflammation. Mucosal Immunol 9(5):1278–1287. https://doi.org/10.1038/mi.2015.129
Shalekoff S, Page-Shipp L, Tiemessen CT (1998) Effects of anticoagulants and temperature on expression of activation markers CD11b and HLA-DR on human leukocytes. Clin Diagn Lab Immunol 5(5):695–702
Plantinga M, Guilliams M, Vanheerswynghels M, Deswarte K, Branco-Madeira F, Toussaint W, Vanhoutte L, Neyt K, Killeen N, Malissen B, Hammad H, Lambrecht BN (2013) Conventional and monocyte-derived CD11b(+) dendritic cells initiate and maintain T helper 2 cell-mediated immunity to house dust mite allergen. Immunity 38(2):322–335. https://doi.org/10.1016/j.immuni.2012.10.016
Bedoret D, Wallemacq H, Marichal T, Desmet C, Quesada Calvo F, Henry E, Closset R, Dewals B, Thielen C, Gustin P, de Leval L, Van Rooijen N, Le Moine A, Vanderplasschen A, Cataldo D, Drion PV, Moser M, Lekeux P, Bureau F (2009) Lung interstitial macrophages alter dendritic cell functions to prevent airway allergy in mice. J Clin Invest 119(12):3723–3738. https://doi.org/10.1172/JCI39717
Yang KD, Liou WY, Lee CS, Chu ML, Shaio MF (1992) Effects of phenobarbital on leukocyte activation: membrane potential, actin polymerization, chemotaxis, respiratory burst, cytokine production, and lymphocyte proliferation. J Leukoc Biol 52(2):151–156
Pecaut MJ, Smith AL, Jones TA, Gridley DS (2000) Modification of immunologic and hematologic variables by method of CO2 euthanasia. Comp Med 50(6):595–602
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Ubags, N.D.J., Suratt, B.T. (2018). Isolation and Characterization of Mouse Neutrophils. In: Alper, S., Janssen, W. (eds) Lung Innate Immunity and Inflammation. Methods in Molecular Biology, vol 1809. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8570-8_4
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8570-8_4
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8569-2
Online ISBN: 978-1-4939-8570-8
eBook Packages: Springer Protocols