High-resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow-derived macrophages

Manman Guo, Anetta Härtlova, Brian D. Dill, Alan R. Prescott, Marek Gierliński, Matthias Trost (Lead / Corresponding author)

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Abstract

Macrophages are important immune cells operating at the forefront of innate immunity by taking up foreign particles and microbes through phagocytosis. The RAW 264.7 cell line is commonly used for experiments in the macrophage and phagocytosis field. However, little is known how its functions compare to primary macrophages. Here, we have performed an in-depth proteomics characterisation of phagosomes from RAW 264.7 and bone marrow-derived macrophages by quantifying more than 2500 phagosomal proteins. Our data indicates that there are significant differences for a large number of proteins including important receptors such as mannose receptor 1 and Siglec-1. Moreover, BMDM phagosomes mature considerably faster by fusion with endosomes and the lysosome when validated using fluorogenic phagocytic assays. We provide a valuable resource for researcher in the field and recommend careful use of the RAW 264.7 cell line when studying phagosome functions. This article is protected by copyright. All rights reserved.

Original languageEnglish
Pages (from-to)3169-3174
Number of pages6
JournalProteomics
Volume15
Issue number18
Early online date5 Feb 2015
DOIs
Publication statusPublished - Sep 2015

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Phagosomes
Macrophages
Proteome
Bone
Phagocytosis
Sialic Acid Binding Ig-like Lectin 1
Cells
Endosomes
Lysosomes
Innate Immunity
Proteomics
Assays
Proteins
Fusion reactions
Research Personnel
Experiments
RAW 264.7 Cells

Cite this

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title = "High-resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow-derived macrophages",
abstract = "Macrophages are important immune cells operating at the forefront of innate immunity by taking up foreign particles and microbes through phagocytosis. The RAW 264.7 cell line is commonly used for experiments in the macrophage and phagocytosis field. However, little is known how its functions compare to primary macrophages. Here, we have performed an in-depth proteomics characterisation of phagosomes from RAW 264.7 and bone marrow-derived macrophages by quantifying more than 2500 phagosomal proteins. Our data indicates that there are significant differences for a large number of proteins including important receptors such as mannose receptor 1 and Siglec-1. Moreover, BMDM phagosomes mature considerably faster by fusion with endosomes and the lysosome when validated using fluorogenic phagocytic assays. We provide a valuable resource for researcher in the field and recommend careful use of the RAW 264.7 cell line when studying phagosome functions. This article is protected by copyright. All rights reserved.",
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year = "2015",
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language = "English",
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High-resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow-derived macrophages. / Guo, Manman; Härtlova, Anetta; Dill, Brian D.; Prescott, Alan R.; Gierliński, Marek; Trost, Matthias (Lead / Corresponding author).

In: Proteomics, Vol. 15, No. 18, 09.2015, p. 3169-3174.

Research output: Contribution to journalArticle

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T1 - High-resolution quantitative proteome analysis reveals substantial differences between phagosomes of RAW 264.7 and bone marrow-derived macrophages

AU - Guo, Manman

AU - Härtlova, Anetta

AU - Dill, Brian D.

AU - Prescott, Alan R.

AU - Gierliński, Marek

AU - Trost, Matthias

N1 - This article is protected by copyright. All rights reserved.

PY - 2015/9

Y1 - 2015/9

N2 - Macrophages are important immune cells operating at the forefront of innate immunity by taking up foreign particles and microbes through phagocytosis. The RAW 264.7 cell line is commonly used for experiments in the macrophage and phagocytosis field. However, little is known how its functions compare to primary macrophages. Here, we have performed an in-depth proteomics characterisation of phagosomes from RAW 264.7 and bone marrow-derived macrophages by quantifying more than 2500 phagosomal proteins. Our data indicates that there are significant differences for a large number of proteins including important receptors such as mannose receptor 1 and Siglec-1. Moreover, BMDM phagosomes mature considerably faster by fusion with endosomes and the lysosome when validated using fluorogenic phagocytic assays. We provide a valuable resource for researcher in the field and recommend careful use of the RAW 264.7 cell line when studying phagosome functions. This article is protected by copyright. All rights reserved.

AB - Macrophages are important immune cells operating at the forefront of innate immunity by taking up foreign particles and microbes through phagocytosis. The RAW 264.7 cell line is commonly used for experiments in the macrophage and phagocytosis field. However, little is known how its functions compare to primary macrophages. Here, we have performed an in-depth proteomics characterisation of phagosomes from RAW 264.7 and bone marrow-derived macrophages by quantifying more than 2500 phagosomal proteins. Our data indicates that there are significant differences for a large number of proteins including important receptors such as mannose receptor 1 and Siglec-1. Moreover, BMDM phagosomes mature considerably faster by fusion with endosomes and the lysosome when validated using fluorogenic phagocytic assays. We provide a valuable resource for researcher in the field and recommend careful use of the RAW 264.7 cell line when studying phagosome functions. This article is protected by copyright. All rights reserved.

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