SpringerLink
Log in

Monocyte Adhesion Assays for Detecting Endothelial Cell Activation in Vascular Inflammation and Atherosclerosis

  • Protocol
  • First Online:
Atherosclerosis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2419))

Abstract

Monocyte adhesion assay, a fluorescence-based method, enables the detection and quantification of monocyte adhesion to endothelial cell (EC) monolayers in vitro and measures EC activation. We describe in this chapter a monocyte adhesion assay based on two published papers from our laboratory that can be effectively used in studying the mechanisms of both pro- and anti-inflammatory cytokines in EC activation. Endothelial cell monolayers are cultured and treated with desired drug, cytokines, or other stimuli and incubated with fluorescently labeled monocytes.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
EUR 44.95
Price includes VAT (Germany)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
EUR 203.29
Price includes VAT (Germany)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
EUR 255.73
Price includes VAT (Germany)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info
Hardcover Book
EUR 353.09
Price includes VAT (Germany)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free ship** worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Roth GA, Johnson C, Abajobir A et al (2017) Global, regional, and national burden of cardiovascular diseases for 10 causes, 1990 to 2015. J Am Coll Cardiol 70(1):1–25. https://doi.org/10.1016/j.jacc.2017.04.052

    Article  PubMed  PubMed Central  Google Scholar 

  2. Gimbrone MA Jr, Garcia-Cardena G (2016) Endothelial cell dysfunction and the pathobiology of atherosclerosis. Circ Res 118(4):620–636. https://doi.org/10.1161/CIRCRESAHA.115.306301

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Mestas J, Ley K (2008) Monocyte-endothelial cell interactions in the development of atherosclerosis. Trends Cardiovasc Med 18(6):228–232. https://doi.org/10.1016/j.tcm.2008.11.004

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Libby P, Pasterkamp G, Crea F, Jang IK (2019) Reassessing the mechanisms of acute coronary syndromes. Circ Res 124(1):150–160. https://doi.org/10.1161/circresaha.118.311098

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Iruela-Arispe ML (2008) Endothelial cell activation. In: Angiogenesis. Springer, Boston. https://doi.org/10.1007/978-0-387-71518-6_3

    Chapter  Google Scholar 

  6. Joseph EK, Green PG, Bogen O, Alvarez P, Levine JD (2013) Vascular endothelial cells mediate mechanical stimulation-induced enhancement of endothelin hyperalgesia via activation of P2X2/3 receptors on nociceptors. J Neurosci 33(7):2849–2859. https://doi.org/10.1523/JNEUROSCI.3229-12.2013

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Branco A, Yoshikawa FSY, Pietrobon AJ, Sato MN (2018) Role of histamine in modulating the immune response and inflammation. Mediat Inflamm 2018:9524075. https://doi.org/10.1155/2018/9524075

    Article  CAS  Google Scholar 

  8. Kelesidis T, Papakonstantinou V, Detopoulou P, Fragopoulou E, Chini M, Lazanas MC, Antonopoulou S (2015) The role of platelet-activating factor in chronic inflammation, immune activation, and comorbidities associated with HIV infection. AIDS Rev 17(4):191–201

    PubMed  PubMed Central  Google Scholar 

  9. Ray CJ, Abbas MR, Coney AM, Marshall JM (2002) Interactions of adenosine, prostaglandins and nitric oxide in hypoxia-induced vasodilatation: in vivo and in vitro studies. J Physiol 544(Pt 1):195–209. https://doi.org/10.1113/jphysiol.2002.023440

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Cybulsky MI, Gimbrone MA Jr (1991) Endothelial expression of a mononuclear leukocyte adhesion molecule during atherogenesis. Science 251(4995):788–791. https://doi.org/10.1126/science.1990440

    Article  CAS  PubMed  Google Scholar 

  11. Li X, Wang L, Fang P, Sun Y, Jiang X, Wang H, Yang XF (2018) Lysophospholipids induce innate immune transdifferentiation of endothelial cells, resulting in prolonged endothelial activation. J Biol Chem 293(28):11033–11045. https://doi.org/10.1074/jbc.RA118.002752

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Lowe DJ, Raj K (2015) Quantitation of endothelial cell adhesiveness in vitro. J Vis Exp 100:e52924. https://doi.org/10.3791/52924

    Article  Google Scholar 

  13. All figures were created with Biorender.com https://biorender.com/

  14. Cell BIOLABS I (2020) Leukocyte endothelium adhesion assay

    Google Scholar 

  15. Fuss IJ (2009) Isolation of whole mononuclear cells from peripheral blood and cord blood. Curr Protoc Immunol Chapter 7:Unit7.1. https://doi.org/10.1002/0471142735.im0701s85

  16. Mai J, Nanayakkara G, Lopez-Pastrana J, Li X, Li YF, Wang X, Song A, Virtue A, Shao Y, Shan H, Liu F, Autieri MV, Kunapuli SP, Iwakura Y, Jiang X, Wang H, Yang XF (2016) Interleukin-17A promotes aortic endothelial cell activation via transcriptionally and post-translationally activating p38 mitogen-activated protein kinase (MAPK) pathway. J Biol Chem 291(10):4939–4954. https://doi.org/10.1074/jbc.M115.690081

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Sha X, Meng S, Li X, ** H, Maddaloni M, Pascual DW, Shan H, Jiang X, Wang H, Yang XF (2015) Interleukin-35 inhibits endothelial cell activation by suppressing MAPK-AP-1 pathway. J Biol Chem 290(31):19307–19318. https://doi.org/10.1074/jbc.M115.663286

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Brocker C, Thompson D, Matsumoto A, Nebert DW, Vasiliou V (2010) Evolutionary divergence and functions of the human interleukin (IL) gene family. Hum Genomics 5(1):30–55. https://doi.org/10.1186/1479-7364-5-1-30

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgment

This work was supported by NIH R01 grants to XY.

Author information

Author notes

  1. Fatma Saaoud and Stephanie Yu shared second authorship.

Authors and Affiliations

  1. Cardiovascular Research Center, Lewis Katz School of Medicine at Temple University, Philadelphia, PA, USA

    Keman Xu, Fatma Saaoud, Charles Drummer IV, Ying Shao, Yu Sun, Yifan Lu, **aohua Jiang & **aofeng Yang

  2. Northeastern University, Boston, MA, USA

    Stephanie Yu

  3. Department of Medicine, Sidney Kimmel Medical College, Thomas Jefferson University, Philadelphia, PA, USA

    Jianxin Sun

  4. Metabolic Disease Research, Departments of Pharmacology, Microbiology and Immunology, Lewis Katz School of Medicine at Temple University, Philadelphia, PA, USA

    Jun Yu, **aohua Jiang, Hong Wang & **aofeng Yang

  5. Centers for Cardiovascular Research, Inflammation & Lung Research, Metabolic Disease Research, and Thrombosis Research, Lewis Katz School of Medicine at Temple University, Philadelphia, PA, USA

    **aofeng Yang

Authors
  1. Keman Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fatma Saaoud
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Stephanie Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Charles Drummer IV
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ying Shao
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yu Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yifan Lu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jianxin Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jun Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. **aohua Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Hong Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. **aofeng Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to **aofeng Yang .

Editor information

Editors and Affiliations

  1. School of Biosciences, Cardiff University, Cardiff, UK

    Dipak Ramji

Rights and permissions

Reprints and permissions

Copyright information

© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Xu, K. et al. (2022). Monocyte Adhesion Assays for Detecting Endothelial Cell Activation in Vascular Inflammation and Atherosclerosis. In: Ramji, D. (eds) Atherosclerosis. Methods in Molecular Biology, vol 2419. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1924-7_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1924-7_10

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1923-0

  • Online ISBN: 978-1-0716-1924-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation