Quantitative GPCR and ion channel transcriptomics in primary alveolar macrophages and macrophage surrogates

BMC Immunology

Toby C Kent¹  Martin Gosling¹  Paul D Wright¹  Lindsay Fawcett¹  Paul J Groot-Kormelink¹ 

[1] Respiratory Disease Area, Novartis Institutes for Biomedical Research, Horsham, RH12 5AB, UK

关键词: High-throughput;    Arrays;    TaqMan;    Microfluidics;    COPD;   

Others  :  1077871
DOI  :  10.1186/1471-2172-13-57

 received in 2012-07-02, accepted in 2012-10-24,  发布年份 2012

【 摘 要 】

Background

Alveolar macrophages are one of the first lines of defence against invading pathogens and play a central role in modulating both the innate and acquired immune systems. By responding to endogenous stimuli within the lung, alveolar macrophages contribute towards the regulation of the local inflammatory microenvironment, the initiation of wound healing and the pathogenesis of viral and bacterial infections. Despite the availability of protocols for isolating primary alveolar macrophages from the lung these cells remain recalcitrant to expansion in-vitro and therefore surrogate cell types, such as monocyte derived macrophages and phorbol ester-differentiated cell lines (e.g. U937, THP-1, HL60) are frequently used to model macrophage function.

Methods

The availability of high throughput gene expression technologies for accurate quantification of transcript levels enables the re-evaluation of these surrogate cell types for use as cellular models of the alveolar macrophage. Utilising high-throughput TaqMan arrays and focussing on dynamically regulated families of integral membrane proteins, we explore the similarities and differences in G-protein coupled receptor (GPCR) and ion channel expression in alveolar macrophages and their widely used surrogates.

Results

The complete non-sensory GPCR and ion channel transcriptome is described for primary alveolar macrophages and macrophage surrogates. The expression of numerous GPCRs and ion channels whose expression were hitherto not described in human alveolar macrophages are compared across primary macrophages and commonly used macrophage cell models. Several membrane proteins known to have critical roles in regulating macrophage function, including CXCR6, CCR8 and TRPV4, were found to be highly expressed in macrophages but not expressed in PMA-differentiated surrogates.

Conclusions

The data described in this report provides insight into the appropriate choice of cell models for investigating macrophage biology and highlights the importance of confirming experimental data in primary alveolar macrophages.

【 授权许可】

   
2012 Groot-Kormelink et al.; licensee BioMed Central Ltd.

【 图 表 】

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Figure 2.

【 参考文献 】

• [1]Global Strategy for the Diagnosis, Management and Prevention of COPD: Global Initiative for Chronic Obstructive Lung Disease (GOLD). 2011. Available from: http://www.goldcopd.org/ webcite
• [2]Barnes PJ: Alveolar macrophages as orchestrators of COPD. COPD 2004, 1:59-70.
• [3]Ebert RH, Florey HW: The extravascular development of the monocyte observed in vivo. Br J Exp Pathol 1939, 20:342-356.
• [4]Daigneault M, Preston JA, Marriott HM, Whyte MKB, Dockrell DH: The Identification of Markers of Macrophage Differentiation in PMA-Stimulated THP-1 Cells and Monocyte-Derived Macrophages. PLoS One 2010, 5:e8668.
• [5]Overington JP, Al-Lazikani B, Hopkins AL: How many drug targets are there? Nat Rev Drug Discov 2006, 5:993-996.
• [6]Rosati B, McKinnon D: Regulation of ion channel expression. Circ Res 2004, 94:874-883.
• [7]Maurel B, Le Digarcher A, Dantec C, Journot L: Genome-wide profiling of G protein-coupled receptors in cerebellar granule neurons using high-throughput, real-time PCR. BMC Genomics 2011, 12:241. BioMed Central Full Text
• [8]Demolombe S, Marionneau C, Le Bouter S, Charpentier F, Escande D: Functional genomics of cardiac ion channel genes. Cardiovasc Res 2005, 67:438-447.
• [9]Lattin J, Zidar DA, Schroder K, Kellie S, Hume DA: G-protein-coupled receptor expression, function and signaling in macrophages. J Leukoc Biol 2007, 82:16-32.
• [10]Newburger PE, Baker RD, Hansen SL, Duncan RA, Greenberger JS: Functionally deficient differentiation of HL-60 promyelocytic leukemia cells induced by phorbol myristate acetate. Cancer Res 1981, 41:1861-1865.
• [11]Devosse T, Guillabert A, D'Haene N, Berton A, De Nadai P, Noel S, Brait M, Franssen J-D, Sozzani S, Salmon I, Parmentier M: Formyl peptide receptor-like 2 is expressed and functional in plasmacytiod dendritic cells, tissue-specific macrophage subpopulations, and eosinophils. J Immunol 2009, 182:4974-4984.
• [12]Alvarez DF, King JA, Weber D, Addison E, Liedtke W, Townsley MI: Transient receptor potential vanilloid 4-mediated disruption of the alveolar septal barrier: a novel mechanism of acute lung injury. Circ Res 2006, 99:988-995.
• [13]Hamanaka K, Jian MY, Townsley MI, King JA, Liedtke W, Weber DS, Eyal FG, Clapp MM, Parker JC: TRPV4 channels augment macrophage activation and ventilator-induced lung injury. Am J Physiol Lung Cell Mol Physiol 2010, 299:L353-L362.
• [14]Insel PA, Snead A, Murray F, Zhang L, Yokouchi H, Katakia T, Kwon O, Dimucci D, Wilderman A: GPCR expression in tissues and cells: are the optimal receptors being used as drug targets? Br J Pharmacol 2012, 165:1613-1616.
• [15]Myrvik Q, Leake ES, Fariss B: Studies on pulmonary alveolar macrophages from the normal rabbit: a technique to procure them in a high state of purity. J Immunol 1961, 86:128-132.
• [16]Burge S, Wedzicha JA: COPD exacerbations: definitions and classifications. Eur Respir J Suppl 2003, 41:46s-53s.
• [17]Berenson CS, Garlipp MA, Grove LJ, Maloney J, Sethi S: Impaired phagocytosis of nontypeable Haemophilus influenzae by human alveolar macrophages in chronic obstructive pulmonary disease. J Infect Dis 2006, 194:1375-1384.
• [18]Taylor AE, Finney-Hayward TK, Quint JK, Thomas CM, Tudhope SJ, Wedzicha JA, Barnes PJ, Donnelly LE: Defective macrophage phagocytosis of bacteria in COPD. Eur Respir J 2010, 35:1039-1047.
• [19]Link TM, Park U, Vonakis BM, Raben DM, Soloski MJ, Caterina MJ: TRPV2 has a pivotal role in macrophage particle binding and phagocytosis. Nat Immunol 2010, 11:232-239.
• [20]Malik M, Jividen K, Padmakumar VC, Cataisson C, Li L, Lee J, Howard OM, Yuspa SH: Inducible NOS-induced chloride intracellular channel 4 (CLIC4) nuclear translocation regulates macrophage deactivation. PNAS 2012, 109:6130-6135.
• [21]Trebst C, Staugaitis SM, Kivisäkk P, Mahad D, Cathcart MK, Tucky B, Wei T, Rani MR, Horuk R, Aldape KD, Pardo CA, Lucchinetti CF, Lassmann H, Ransohoff RM: CC chemokine receptor 8 in the central nervous system is associated with phagocytic macrophages. Am J Pathol 2003, 162:427-438.
• [22]Reimer MK, Brange C, Rosendahl A: CCR8 signaling influences Toll-like receptor 4 responses in human macrophages in inflammatory diseases. Clin Vaccine Immunol 2011, 18:2050-2059.
• [23]Collman RG, Perno CF, Crowe SM, Stevenson M, Montaner LJ: HIV and cells of macrophage/dendritic lineage and other non-T cell reservoirs: new answers yield new questions. J Leukoc Biol 2003, 74:631-634.
• [24]Sharova N, Swingler C, Sharkey M, Stevenson M: Macrophages archive HIV-1 virions for dissemination in trans. EMBO J 2005, 24:2481-2489.
• [25]Worgall S, Connor R, Kaner RJ, Fenamore E, Sheridan K, Singh R, Crystal RG: Expression and use of human immunodeficiency virus type 1 coreceptors by human alveolar macrophages. J Virol 1999, 73:5865-5874.
• [26]Feng Y, Broder CC, Kennedy PE, Berger EA: HIV-1 entry cofactor: functional cDNA cloning of a seven-transmembrane, G protein-coupled receptor. Science 1996, 272:872-877.
• [27]Alkhatib G, Combadiere C, Broder CC, Feng Y, Kennedy PE, Murphy PM, Berger EA: CC CKR5: a RANTES, MIP-1alpha, MIP-1beta receptor as a fusion cofactor for macrophage-tropic HIV-1. Science 1996, 272:1955-1958.
• [28]Kootstra NA, Schuitemaker H: Determination of cell tropism of HIV-1. Methods Mol Biol 2005, 304:317-325.
• [29]Lee CH, Lee GC, Chan YJ, Chiou CJ, Ahn JH, Hayward GS: Factors affecting human cytomegalovirus gene expression in human monocyte cell lines. Mol Cells 1999, 9:37-44.
• [30]Schuitemaker H, Kootstra NA, Groenink M, De Goede RE, Miedema F, Tersmette M: Differential tropism of clinical HIV-1 isolates for primary monocytes and promonocytic cell lines. AIDS Res Hum Retroviruses 1992, 8:1679-1682.
• [31]Ackerman JJ, Duerre JA: Long-term culturing of TPA-induced differentiated HL-60 cells results in increased levels of lytic enzymes. Exp Cell Res 1989, 183:353-360.
• [32]Mosser DM, Edwards JP: Exploring the full spectrum of macrophage activation. Nat Rev Immunol 2008, 8:958-969.
• [33]Gordon S, Taylor PR: Monocyte and macrophage heterogeneity. Nature Rev Immunol 2005, 5:953-964.
• [34]Nibbering PH, van der Heide A, Van Furth R: Macrophages in bronchoalveolar lavage fluid are not representative of macrophages in granulomas of the lungs of BCG-infected mice. J Pathol 1989, 157:253-261.
• [35]Armstrong J, Sargent C, Singh D: Glucocorticoid sensitivity of lipopolysaccharide-stimulated chronic obstructive pulmonary disease alveolar macrophages. Clin Exp Immunol 2009, 158:74-83.