Skip to main content

Advertisement

SpringerLink
Log in

Optimized flow cytometry protocol for analysis of surface expression of interleukin-1 receptor types I and II

  • Method in Cell Science
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

The biological effects of interleukin (IL)-1 are realized through binding to specific membrane-bound receptors. The efficiency of IL-1 action depends on the number of receptors on the cell. We determined the percentage of cells that express IL-1 receptor type I (IL-1RI) and IL-1 receptor type II (IL-1RII) by flow cytometry using phycoerythrin (PE)-labelled antibodies to the IL-1Rs, and the mean absolute number of membrane-bound IL-1Rs per cell using QuantiBRITE PE calibration beads. We showed that different subpopulations of immunocompetent cells expressed different numbers of molecules of membrane-bound IL-1RI and IL-1RII. We also established that when cells were stimulated with bacterial lipopolysaccharide, there was a significant increase in the number of IL-1RI expressed, and a significant decrease in the mean number of IL-1RII molecules per cell. Determination of the mean number of membrane-bound IL-1R molecules using this protocol enables us to obtain precise and reproducible data that are necessary for full evaluation of expression levels.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Böyum A (1968) Separation of leukocytes from blood and bone marrow. Introduction. Scand J Clin Lab Invest Suppl 97:7

  • Colotta F, Re F, Muzio M, Bertini R, Polentarutti N, Sironi M, Giri JG, Dower SK, Sims JE, Mantovani A (1993) Interleukin-1 type II receptor: a decoy target for IL-1 that regulated by IL-4. Science 261:472–475

    Article  CAS  Google Scholar 

  • D’hautcourt JL (2002) Quantitative flow cytometric analysis of membrane antigen expression. Curr Protoc Cytom 6.12.1–6.12.22

  • Dinarello CA (1991) Interleukin-1 and interleukin-1 antagonism. Blood 77:1627–1652

    CAS  Google Scholar 

  • Dinarello CA (2001) IL-1 receptor type I. In: Oppenheim JJ, Feldmann M, Durum SK, Hirano T, Vilcek J, Nicola NA (eds) Cytokine reference: A compendium of cytokines and other mediators of host defense. Academic Press, London, pp 1587–1600

    Google Scholar 

  • Dinarello CA (2010) IL-1: discoveries, controversies and future directions. Eur J Immunol 40:599–606

    Article  CAS  Google Scholar 

  • Donati D, Degiannis D, Mazzola E, Gastaldi L, Raskova J, Raska KJ, Camussi G (1997) Interleukin-1 receptors and receptor antagonist in haemodialysis. Nephrol Dial Transplant 12:111–118

    Article  CAS  Google Scholar 

  • Dower SK, Kronheim SR, March CJ, Conion PJ, Hopp TP, Gillis S, Urdal DL (1985) Detection and characterization of high affinity plasma membrane receptors for human interleukin 1. J Exp Med 162:501–515

    Article  CAS  Google Scholar 

  • Gudipaty L, Humphreys BD, Buell G, Dubyak GR (2001) Regulation of P2X(7) nucleotide receptor function in human monocytes by extracellular ions and receptor density. Am J Physiol Cell Physiol 280:C943–C953

    CAS  Google Scholar 

  • Jasper GA, Arun I, Venzon D, Kreitman RJ, Wayne AS, Yuan CM, Marti GE, Stetler-Stevenson M (2011) Variables affecting the quantitation of CD22 in neoplastic B cells. Cytometry B Clin Cytom 80:83–90

    Google Scholar 

  • Mantovani A, Locati M, Vecchi A, Sozzani S, Allavena P (2001) Decoy receptors: a strategy to regulate inflammatory cytokines and chemokines. Trends Immunol 22:328–336

    Article  CAS  Google Scholar 

  • Matsushima K, Akahoshi T, Yamada M, Furutani Y, Oppenheim JJ (1986) Properties of a specific interleukin 1 (IL1) receptor on human Epstein Barr virus-transformed B lymphocytes: identity of a receptor for IL 1-alpha and IL 1-beta. J Immunol 136:4496–4502

    CAS  Google Scholar 

  • Moraga I, Harari D, Schreiber G, Uzé G, Pellegrini S (2009) Receptor density is key to the Alpha2/Beta interferon differential activities. Mol Cell Biol 29:4778–4787

    Article  CAS  Google Scholar 

  • Orlova DY, Borisov VI, Kozhevnikov VS, Maltscev VP, Chernyshev AV (2011) Distribution function approach to the study of the kinetics of IgM antibody binding to FcγRIIIb (CD16b) receptors on neutrophils by flow cytometry. J Theor Biol 290:1–6

    Article  CAS  Google Scholar 

  • Pannu KK, Joe ET, Iyer SB (2001) Performance evaluation of QuantiBRITE phycoerythrin beads. Cytometry 45:250–258

    Article  CAS  Google Scholar 

  • Penton-Rol G, Orlando S, Polentarutti N, Bernasconi S, Muzio M, Introna M, Mantovani A (1999) Bacterial lipopolysaccharide causes rapid shedding, followed by inhibition of mRNA expression, of the IL-1 type II receptor, with concomitant up-regulation of the type I receptor and induction of incompletely spliced transcripts. J Immunol 162:2931–2938

    CAS  Google Scholar 

  • Reynes J, Portales P, Segondy M, Baillat V, André P, Réant B, Avinens O, Couderc G, Benkirane M, Clot J, Eliaou JF, Corbeau P (2000) CD4 + T cell surface CCR5 density as a determining factor of virus load in persons infected with human immunodeficiency virus type 1. J Infect Dis 181:927–933

    Article  CAS  Google Scholar 

  • Rossmann ED, Lenkei R, Lundin J, Mellstedt H, Osterborg A (2007) Performance of calibration standards for antigen quantitation with flow cytometry in chronic lymphocytic leukemia. Cytometry B Clin Cytom 72:450–457

    Google Scholar 

  • Saccani S, Polentarutti N, Penton-Rol G, Sims JE, Mantovani A (1998) Divergent effects of LPS on expression of IL-1 receptor family members in mononuclear phagocytes in vitro and in vivo. Cytokine 10:773–780

    Article  CAS  Google Scholar 

  • Sims JE, Gayle MA, Slack JL, Alderson MR, Bird TA, Giri JG, Colotta F, Re F, Mantovani A, Shanebeck K, Grabstein KH, Dower SK (1993) Interleukin 1 signaling occurs exclusively via the type I receptor. Proc Natl Acad Sci USA 90:6155–6159

    Article  CAS  Google Scholar 

  • Stadnichuk IN (1990) Phycobiliproteins. VINITI, Moscow

    Google Scholar 

  • Stylianou E, O’Neill LA, Rawlinson L, Edbrooke MR, Woo P, Saklatvala J (1992) Interleukin 1 induces NF-kappa B through its type I but not its type II receptor in lymphocytes. J Biol Chem 267:15836–15841

    CAS  Google Scholar 

  • Takeuchi M, Nagai S, Tsutumi T, Mio T, Izumi T (1999) The number of interleukin 1 receptors on lung fibroblasts in patients with idiopathic pulmonary fibrosis. Respiration 66:236–241

    Article  CAS  Google Scholar 

  • Wang L, Abbasi F, Jasper GA, Kreitman RJ, Liewehr DJ, Marti GE, Stetler-Stevenson M (2011) Variables in the quantification of CD4 in normals and hairy cell leukemia patients. Cytometry B Clin Cytom 80:51–56

    Google Scholar 

Download references

Acknowledgments

This study was supported by the Federal Target Program “Scientific and scientific-pedagogical personnel of innovative Russia” for 2009–2013 (state contract no. 02.740.11.0707).

Author information

Authors and Affiliations

  1. Laboratory of Molecular Immunology, Federal State Budgetary Institution “Research Institute of Clinical Immunology”, Russian Academy of Medical Sciences Siberian Branch, 14, Yadrincevskaja str., 630099, Novosibirsk, Russia

    Filipp Filippovich Vasilyev, Julia Anatolievna Lopatnikova & Sergey Vitalievich Sennikov

Authors
  1. Filipp Filippovich Vasilyev
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Julia Anatolievna Lopatnikova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sergey Vitalievich Sennikov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sergey Vitalievich Sennikov.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vasilyev, F.F., Lopatnikova, J.A. & Sennikov, S.V. Optimized flow cytometry protocol for analysis of surface expression of interleukin-1 receptor types I and II. Cytotechnology 65, 795–802 (2013). https://doi.org/10.1007/s10616-013-9546-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-013-9546-6

Keywords

Search

Navigation