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Cell identification and isolation on the basis of cytokine secretion: A novel tool for investigating immune responses

Nature Medicine volume 7, pages 373–376 (2001)Cite this article

The function and differentiation state of many cell types is more accurately defined by the molecules they secrete rather than by their surface phenotype. For example, the T-helper (CD4+) lymphocyte subsets, Th1 and Th2, are best defined by their production of interferon-γ (IFN-γ) and interleukin (IL)-4, respectively1. Moreover, recently described regulatory subsets of T cells are characterized by the secretion of transforming growth factor (TGF)-β and IL-10 (refs. 2,3). Investigations into the function of such cells and their potential uses in therapy require cytokine-based identification methods that do not compromise cell viability or physiological state. Here we describe a novel method that allows precise identification of cytokine secretion patterns in viable cells. The method, where the cells are encapsulated in a permeable antibody-capture matrix, is based on technologies previously established to isolate secretory cells4,5 and have been used for the selection of antibody-secreting B-cell hybridomas6. The adaptations that we have made to the technique allow it to be easily and conveniently used to detect cytokine production and sort cells on the basis of secretion patterns. Here we used the new technique in conjunction with flow cytometry to distinguish lymphocytes secreting IFN-γ, IL-4, IL-10 or TGF-β. We present evidence that the method neither alters the secretion patterns of the cells, nor affects their response to conventional stimulation following removal of the agarose matrix. The ability to combine this technique with cell-surface staining indicates its potential as a powerful tool in understanding the role of T-cell subsets in regulating the immune system and also makes it applicable in isolating a broad range of other cell types.

Characterization of the capture matrix for secreted cytokines

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Figure 1: Cytokine detection using gel microdrop micro-encapsulation.
Figure 2: Microencapsulation allows the detection of cytokine production.
Figure 3: Microencapsulation does not alter cytokine secretion by human PBMCs.

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Acknowledgements

We thank S. Sreckovic for assistance with cell sorting and Ian Patterson for the V69C5 cells. N.A.W. is a Wellcome Trust Fellow.

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Authors and Affiliations

  1. Department of Pathology and Microbiology, University of Bristol, School of Medical Sciences, University Walk, Bristol, UK

    Victor Turcanu & Neil A. Williams

  2. Department of Pediatrics, Imperial College of Medicine, Norfolk PLace, London, UK

    Victor Turcanu

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  1. Victor Turcanu
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  2. Neil A. Williams
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Correspondence to Neil A. Williams.

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Turcanu, V., Williams, N. Cell identification and isolation on the basis of cytokine secretion: A novel tool for investigating immune responses. Nat Med 7, 373–376 (2001). https://doi.org/10.1038/85533

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