High-sensitivity immunofluorescence for detection of the pro- and anti-inflammatory cytokines gamma interferon and interleukin-10 on the surface of cytokine-secreting cells (original) (raw)

Nature Medicine volume 6, pages 107–110 (2000)Cite this article

Immunofluorescence-based analyses have become a powerful tool to characterize cells according to the expression of specific proteins. However, the sensitivity of the technology is limited because of the low number of fluorochromes that can be conjugated to one staining antibody. A minimum of several thousand antigens per cell is required for discrimination between positive and negative cells1, although functionally important molecules, for example, many cytokine or hormone receptors2, or the pre-B- and pre-T-cell receptors3,4, are expressed at much lower copy numbers. Here, we show that antibody-conjugated magnetofluorescent liposomes, which can increase fluorescence signal intensity 100-fold to 1,000-fold compared with conventional methods5, allow the detection of 50–100 target molecules per cell. Using this technique, we have characterized the expression of a new surface-bound form of interleukin (IL)-10 and a recently described surface form of interferon (IFN)-γ (ref. 6), which are almost undetectable by conventional technologies, although there is indirect evidence for the existence of physiologically active surface-bound IFN-γ from functional studies7,8. In addition to this functional role, surface IFN-γ and IL-10 represent the first natural surface markers for the unambiguous identification and isolation of viable IFN-γ and IL-10-producing cells, the two essential regulators of inflammatory and anti-inflammatory immune responses.

Characteristics of magnetofluorescent liposomes

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Acknowledgements

We thank R. Christine, D. Kunkel and C. Raulfs for critical reading of the manuscript.

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Authors and Affiliations

  1. Deutsches Rheuma-Forschungszentrum Berlin, Hannoversche Str. 27, Berlin, 10115, Germany
    Alexander Scheffold, Luzie Reiners-Schramm, Roland Lauster & Andreas Radbruch
  2. Miltenyi Biotec GmbH, Bergisch Gladbach, Germany
    Mario Assenmacher

Authors

  1. Alexander Scheffold
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  2. Mario Assenmacher
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  3. Luzie Reiners-Schramm
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  4. Roland Lauster
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  5. Andreas Radbruch
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Correspondence toAlexander Scheffold.

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Scheffold, A., Assenmacher, M., Reiners-Schramm, L. et al. High-sensitivity immunofluorescence for detection of the pro- and anti-inflammatory cytokines gamma interferon and interleukin-10 on the surface of cytokine-secreting cells.Nat Med 6, 107–110 (2000). https://doi.org/10.1038/71441

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