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Detecting Secreted Analytes from Immune Cells: An Overview of Technologies

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The Tumor Microenvironment

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1458))

Abstract

The tumor microenvironment is largely shaped by secreted factors and infiltrating immune cells and the nature of this environment can profoundly influence tumor growth and progression. As such, there is an increasing need to identify and quantify secreted factors by tumor cells, tumor-associated cells, and infiltrating immune cells. To meet this need, the dynamic range of immunoassays such as ELISAs and ELISpots have been improved and the scope of reagents commercially available has been expanded. In addition, new bead-based and membrane-based screening arrays have been developed to allow for the simultaneous detection of multiple analytes in one sample. Similarly, the optimization of intracellular staining for flow cytometry now allows for the quantitation of multiple cytokines from either a purified cell population or a complex mixed cell suspension. Herein, we review the rapidly evolving technologies that are currently available to detect secreted analytes. Emphasis is placed on discussing the advantages and disadvantages of these assays and their applications.

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References

  1. McAllister SS, Weinberg RA (2014) The tumour-induced systemic environment as a critical regulator of cancer progression and metastasis. Nat Cell Biol 16:717–727

    Article  CAS  PubMed  Google Scholar 

  2. Chai EZ, Siveen KS, Shanmugam MK, Arfuso F, Sethi G (2015) Analysis of the intricate relationship between chronic inflammation and cancer. Biochem J 468:1–15

    Article  CAS  PubMed  Google Scholar 

  3. Gajewski TF, Schreiber H, Fu YX (2013) Innate and adaptive immune cells in the tumor microenvironment. Nat Immunol 14:1014–1022

    Article  CAS  PubMed Central  Google Scholar 

  4. Mahmoud S, Lee A, Ellis I, Green A (2012) CD8(+) T lymphocytes infiltrating breast cancer: a promising new prognostic marker? Oncoimmunology 1:364–365

    Article  PubMed  PubMed Central  Google Scholar 

  5. Galon J et al (2012) The immune score as a new possible approach for the classification of cancer. J Transl Med 10:1

    Article  PubMed  PubMed Central  Google Scholar 

  6. Motz GT, Coukos G (2013) Deciphering and reversing tumor immune suppression. Immunity 39:61–73

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Landskron G, De la Fuente M, Thuwajit P, Thuwajit C, Hermoso MA (2014) Chronic inflammation and cytokines in the tumor microenvironment. J Immunol Res 2014:149185

    Article  PubMed  PubMed Central  Google Scholar 

  8. Wang D, DuBois RN (2015) Immunosuppression associated with chronic inflammation in the tumor microenvironment. Carcinogenesis 36:1085–1093

    Article  CAS  PubMed  Google Scholar 

  9. Floros T, Tarhini AA (2015) Anticancer cytokines: biology and clinical effects of interferon-alpha2, interleukin (IL)-2, IL-15, IL-21, and IL-12. Sem Oncol 42:539–548

    Article  CAS  Google Scholar 

  10. Wu AA, Drake V, Huang HS, Chiu S, Zheng L (2015) Reprogramming the tumor microenvironment: tumor-induced immunosuppressive factors paralyze T cells. Oncoimmunology 4:e1016700

    Article  PubMed  PubMed Central  Google Scholar 

  11. Mittal SK, Roche PA (2015) Suppression of antigen presentation by IL-10. Curr Opin Immunol 34:22–27

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Dennis KL, Blatner NR, Gounari F, Khazaie K (2013) Current status of interleukin-10 and regulatory T-cells in cancer. Curr Opin Oncol 25:637–645

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Liu J, Cao X (2015) Regulatory dendritic cells in autoimmunity: a comprehensive review. J Autoimmun 63:1–12

    Article  CAS  PubMed  Google Scholar 

  14. Benencia F, Muccioli M, Alnaeeli M (2014) Perspectives on reprograming cancer-associated dendritic cells for anti-tumor therapies. Front Oncol 4:72

    Article  PubMed  PubMed Central  Google Scholar 

  15. Schuerwegh AJ, Stevens WJ, Bridts CH, De Clerck LS (2001) Evaluation of monensin and brefeldin A for flow cytometric determination of interleukin-1 beta, interleukin-6, and tumor necrosis factor-alpha in monocytes. Cytometry 46:172–176

    Article  CAS  PubMed  Google Scholar 

  16. Nylander S, Kalies I (1999) Brefeldin A, but not monensin, completely blocks CD69 expression on mouse lymphocytes: efficacy of inhibitors of protein secretion in protocols for intracellular cytokine staining by flow cytometry. J Immunol Methods 224:69–76

    Article  CAS  PubMed  Google Scholar 

  17. O'Neil-Andersen NJ, Lawrence DA (2002) Differential modulation of surface and intracellular protein expression by T cells after stimulation in the presence of monensin or brefeldin A. Clin Diagn Lab Immunol 9:243–250

    PubMed  PubMed Central  Google Scholar 

  18. Zola H (2004) High-sensitivity immunofluorescence/flow cytometry: detection of cytokine receptors and other low-abundance membrane molecules. Curr Prot Cytom Chapter 6:Unit 6.3

    Google Scholar 

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Acknowledgements

We acknowledge the support of the Leukemia and Lymphoma Society of Canada and the Canadian Cancer Society Research Institute to K.A.P., and the CIHR and NSERC for funding to C.K.

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Correspondence to Connie M. Krawczyk .

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© 2016 Springer Science+Business Media New York

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Pike, K.A., Hui, C., Krawczyk, C.M. (2016). Detecting Secreted Analytes from Immune Cells: An Overview of Technologies. In: Ursini-Siegel, J., Beauchemin, N. (eds) The Tumor Microenvironment. Methods in Molecular Biology, vol 1458. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3801-8_9

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  • DOI: https://doi.org/10.1007/978-1-4939-3801-8_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3799-8

  • Online ISBN: 978-1-4939-3801-8

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