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Classification of the Immune Composition in the Tumor Infiltrate

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Single Cell Methods

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1979))

Abstract

Flow cytometry is one of the most suitable techniques for analyzing and classifying different cell suspensions derived from blood or others compartments. The characterization of all different cellular subtypes is made with different antibodies that detect surface or intracytoplasmic antigens. Here we describe the technique to thoroughly characterize immune cells from tumor infiltrates and proceed to isolation using single-cell sorting.

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References

  1. Jaye DL, Bray RA, Gebel HM, Harris WAC, Waller EK (2012) Translational applications of flow cytometry in clinical practice. J Immunol 188(10):4715–4719

    CAS  PubMed  Google Scholar 

  2. Ma W, Gilligan BM, Yuan J, Li T (2016) Current status and perspectives in translational biomarker research for PD-1/PD-L1 immune checkpoint blockade therapy. J Hematol Oncol 9:47

    PubMed  PubMed Central  Google Scholar 

  3. Koonce NA, Juratli MA, Cai C, Sarimollaoglu M, Menyaev YA, Dent J, Quick CM, Dings RPM, Nedosekin D, Zharov V, Griffin RJ (2017) Real-time monitoring of circulating tumor cell (CTC) release after nanodrug or tumor radiotherapy using in vivo flow cytometry. Biochem Biophys Res Commun 492(3):507–512

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Menck K, Bleckmann A, Wachter A, Hennies B, Ries L, Schulz M, Balkenhol M, Pukrop T, Schatlo B, Rost U, Wenzel D, Klemm F, Binder C (2017) Characterisation of tumour-derived microvesicles in cancer patients’ blood and correlation with clinical outcome. J Extracell Vesicles 6(1):1340745

    PubMed  PubMed Central  Google Scholar 

  5. Burger D, Oleynik P (2017) Isolation and characterization of circulating microparticles by flow cytometry. Methods Mol Biol 1527:271–281

    CAS  PubMed  Google Scholar 

  6. McLaughlin BE, Baumgarth N, Bigos M, Roederer M, De Rosa SC, Altman JD, Nixon DF, Ottinger J, Oxford C, Evans TG, Asmuth DM (2008) Nine-color flow cytometry for accurate measurement of T cell subsets and cytokine responses. Part I: Panel design by an empiric approach. Cytometry A 73(5):400–410

    PubMed  Google Scholar 

  7. Basu S, Campbell HM, Dittel BN, Ray A (2010) Purification of specific cell population by fluorescence activated cell sorting (FACS). J Vis Exp (41):e1546

    Google Scholar 

  8. Herzenberg LA, De Rosa SC, Herzenberg LA (2000) Monoclonal antibodies and the FACS: complementary tools for immunobiology and medicine. Immunol Today 21:383–390

    CAS  PubMed  Google Scholar 

  9. Tung JW, Heydari K, Tirouvanziam R, Sahaf B, Parks DR, Herzenberg LA, Herzenberg LA (2007) Modern flow cytometry: a practical approach. Clin Lab Med 27(3):453–468

    PubMed  PubMed Central  Google Scholar 

  10. Brusa D, Carletto S, Cucchiarale G, Gontero P, Greco A, Simone M, Ferrando U, Tizzani A, Matera L (2011) Prostatectomy restores the maturation competence of blood dendritic cell precursors and reverses the abnormal expansion of regulatory T lymphocytes. Prostate 71:344–352

    CAS  PubMed  Google Scholar 

  11. Gorosito Serrán M, Fiocca Vernengo F, Beccaria CG, Acosta Rodriguez EV, Montes CL, Gruppi A (2015) The regulatory role of B cells in autoimmunity, infections and cancer: perspectives beyond IL10 production. FEBS Lett 589(22):3362–3369

    PubMed  Google Scholar 

  12. Allavena P, Chieppa M, Bianchi G, Solinas G, Fabbri M, Laskarin G, Mantovani A (2010) Engagement of the mannose receptor by tumoral mucins activates an immune suppressive phenotype in human tumor-associated macrophages. Clin Dev Immunol 2010:547179. https://doi.org/10.1155/2010/547179

    Article  CAS  PubMed  Google Scholar 

  13. Brusa D, Simone M, Gontero P, Spadi R, Racca P, Micari J, Degiuli M, Carletto S, Tizzani A, Matera L (2013) Circulating immunosuppressive cells of prostate cancer patients before and after radical prostatectomy: Profile comparison. Int J Urol 20:971–978

    CAS  PubMed  Google Scholar 

  14. Hu P, Zhang Z, Xin H, Deng G (2016) Single cell isolation and analysis. Front Cell Dev Biol 4:116

    PubMed  PubMed Central  Google Scholar 

  15. Reitsma MJ, Lee BR, Uchida N (2002) Method for purification of human hematopoietic stem cells by flow cytometry. Methods Mol Med 63:59–77

    PubMed  Google Scholar 

  16. Karabinus DS, Marazzo DP, Stern HJ, Potter DA, Opanga CI, Cole ML, Johnson LA, Schulman JD (2014) The effectiveness of flow cytometric sorting of human sperm (MicroSort®) for influencing a child’s sex. Reprod Biol Endocrinol 12:106

    PubMed  PubMed Central  Google Scholar 

  17. Battye FL, Light A, Tarlinton DM (2000) Single cell sorting and cloning. J Immunol Methods 243:25–32

    CAS  PubMed  Google Scholar 

  18. Six A, Mariotti-Ferrandiz ME, Chaara W, Magadan S, Pham HP, Lefranc MP, Mora T, Thomas-Vaslin V, Walczak AM, Boudinot P (2013) The past, present, and future of immune repertoire biology – the rise of next-generation repertoire analysis. Front Immunol 4:413

    PubMed  PubMed Central  Google Scholar 

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Author information

Authors and Affiliations

  1. Flow Cytometry Platform, Institute of Experimental and Clinical Research (IREC), Université Catholique de Louvain (UCL), Brussels, Belgium

    Davide Brusa

  2. Pole of Pharmacology and Therapeutics, Institute of Experimental and Clinical Research (IREC), Medical School, Université Catholique de Louvain (UCL), Brussels, Belgium

    Jean-Luc Balligand

Authors
  1. Davide Brusa
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  2. Jean-Luc Balligand
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Corresponding author

Correspondence to Davide Brusa .

Editor information

Editors and Affiliations

  1. Department of Life Sciences and System Biology, University of Turin, Italian Institute for Genomic Medicine, IIGM Turin, Torino, Italy

    Valentina Proserpio

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Brusa, D., Balligand, JL. (2019). Classification of the Immune Composition in the Tumor Infiltrate. In: Proserpio, V. (eds) Single Cell Methods. Methods in Molecular Biology, vol 1979. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9240-9_19

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  • DOI: https://doi.org/10.1007/978-1-4939-9240-9_19

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9239-3

  • Online ISBN: 978-1-4939-9240-9

  • eBook Packages: Springer Protocols

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