Harmonisation of short-term in vitro culture for the expansion of antigen-specific CD8+ T cells with detection by ELISPOT and HLA-multimer staining

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• 作者：Lindsey Chudley (1)
  Katy J. McCann (1)
  Adam Coleman (1)
  Angelica M. Cazaly (1)
  Nicole Bidmon (2)
  Cedrik M. Britten (2)
  Sjoerd H. van der Burg (3)
  Cecile Gouttefangeas (4)
  Camilla Jandus (5)
  Karoline Laske (4)
  Dominik Maurer (6)
  Pedro Romero (5)
  Helene Schr?der (2)
  Linda F. M. Stynenbosch (3)
  Steffen Walter (6)
  Marij J. P. Welters (3)
  Christian H. Ottensmeier (1) (7)
  
• 关键词：T cell ; In vitro stimulation ; ELISPOT ; Multimer ; Harmonisation ; Inter ; laboratory testing
• 刊名：Cancer Immunology, Immunotherapy
• 出版年：2014
• 出版时间：November 2014
• 年：2014
• 卷：63
• 期：11
• 页码：1199-1211
• 全文大小：1,945 KB
• 参考文献：1. Boaz MJ, Hayes P, Tarragona T, Seamons L, Cooper A, Birungi J, Kitandwe P, Semaganda A, Kaleebu P, Stevens G, Anzala O, Farah B, Ogola S, Indangasi J, Mhlanga P, Van Eeden M, Thakar M, Pujari A, Mishra S, Goonetilleke N, Moore S, Mahmoud A, Sathyamoorthy P, Mahalingam J, Narayanan PR, Ramanathan VD, Cox JH, Dally L, Gill DK, Gilmour J (2009) Concordant proficiency in measurement of T-cell immunity in human immunodeficiency virus vaccine clinical trials by peripheral blood mononuclear cell and enzyme-linked immunospot assays in laboratories from three continents. Clin Vaccine Immunol 16:147-55 CrossRef
  2. Schloot NC, Meierhoff G, Karlsson Faresj? M, Ott P, Putnam A, Lehmann P, Gottlieb P, Roep BO, Peakman M, Tree T (2003) Comparison of cytokine ELISpot assay formats for the detection of islet antigen autoreactive T cells. Report of the third immunology of diabetes society T-cell workshop. J Autoimmun 21:365-76 CrossRef
  3. Scheibenbogen C, Romero P, Rivoltini L, Herr W, Schmittel A, Cerottini JC, Woelfel T, Eggermont AM, Keilholz U (2000) Quantitation of antigen-reactive T cells in peripheral blood by IFNgamma-ELISPOT assay and chromium-release assay: a four-centre comparative trial. J Immunol Methods 244:81-9 CrossRef
  4. Smith SG, Joosten SA, Verscheure V, Pathan AA, McShane H, Ottenhoff TH, Dockrell HM, Mascart F (2009) Identification of major factors influencing ELISpot-based monitoring of cellular responses to antigens from Mycobacterium tuberculosis. PLoS ONE 4:e7972 CrossRef
  5. Cox, J. H., Ferrari, G., Kalams, S. A., Lopaczynski, W., Oden, N., D’souza, M. P., Group, E. C. S (2005) Results of an ELISPOT proficiency panel conducted in 11 laboratories participating in international human immunodeficiency virus type 1 vaccine trials. AIDS Res Hum Retroviruses 21:68-1 CrossRef
  6. Mander A, Gouttefangeas C, Ottensmeier C, Welters MJ, Low L, van der Burg SH, Britten CM (2010) Serum is not required for ex vivo IFN-gamma ELISPOT: a collaborative study of different protocols from the European CIMT Immunoguiding Program. Cancer Immunol Immunother 59:619-27 CrossRef
  7. Janetzki S, Panageas KS, Ben-Porat L, Boyer J, Britten CM, Clay TM, Kalos M, Maecker HT, Romero P, Yuan J, Kast WM, Hoos A (2008) Results and harmonization guidelines from two large-scale international Elispot proficiency panels conducted by the Cancer Vaccine Consortium (CVC/SVI). Cancer Immunol Immunother 57:303-15 CrossRef
  8. Filbert H, Attig S, Bidmon N, Renard BY, Janetzki S, Sahin U, Welters MJ, Ottensmeier C, van der Burg SH, Gouttefangeas C, Britten CM (2013) Serum-free freezing media support high cell quality and excellent ELISPOT assay performance across a wide variety of different assay protocols. Cancer Immunol Immunother 62:615-27 CrossRef
  9. Janetzki S, Price L, Britten CM, van der Burg SH, Caterini J, Currier JR, Ferrari G, Gouttefangeas C, Hayes P, Kaempgen E, Lennerz V, Nihlmark K, Souza V, Hoos A (2010) Performance of serum-supplemented and serum-free media in IFNgamma Elispot Assays for human T cells. Cancer Immunol Immunother 59:609-18 CrossRef
  10. Britten CM, Janetzki S, Ben-Porat L, Clay TM, Kalos M, Maecker H, Odunsi K, Pride M, Old L, Hoos A, Romero P (2009) Harmonization guidelines for HLA-peptide multimer assays derived from results of a large scale international proficiency panel of the Cancer Vaccine Consortium. Cancer Immunol Immunother 58:1701-713 CrossRef
  11. Britten CM, Gouttefangeas C, Welters MJ, Pawelec G, Koch S, Ottensmeier C, Mander A, Walter S, Paschen A, Muller-Berghaus J, Haas I, Mackensen A, Kollgaard T, Thor Straten P, Schmitt M, Giannopoulos K, Maier R, Veelken H, Bertinetti C, Konur A, Huber C, Stevanovic S, Wolfel T, van der Burg SH (2008) The C
• 作者单位：Lindsey Chudley (1)
  Katy J. McCann (1)
  Adam Coleman (1)
  Angelica M. Cazaly (1)
  Nicole Bidmon (2)
  Cedrik M. Britten (2)
  Sjoerd H. van der Burg (3)
  Cecile Gouttefangeas (4)
  Camilla Jandus (5)
  Karoline Laske (4)
  Dominik Maurer (6)
  Pedro Romero (5)
  Helene Schr?der (2)
  Linda F. M. Stynenbosch (3)
  Steffen Walter (6)
  Marij J. P. Welters (3)
  Christian H. Ottensmeier (1) (7)
  
  1. Cancer Sciences Unit, Faculty of Medicine, Experimental Cancer Medicine Centre, Southampton General Hospital, University of Southampton, Tremona Road, Southampton, SO16 6YD, UK
  2. Translational Oncology, University Medical Center, Johannes-Gutenberg University GmbH, Mainz, Germany
  3. Department of Clinical Oncology, Leiden University Medical Centre, Leiden, The Netherlands
  4. Department of Immunology, Institute for Cell Biology, Eberhard-Karls University, Tübingen, Germany
  5. Translational Tumour Immunology, Ludwig Institute for Cancer Research, Lausanne, Switzerland
  6. Immatics Biotechnologies GmbH, Tübingen, Germany
  7. Somers Cancer Research Building (Mailpoint 824), Cancer Sciences Unit, Faculty of Medicine, Southampton General Hospital, University of Southampton, Tremona Road, Southampton, SO16 6YD, UK
  
• ISSN：1432-0851

文摘

Ex vivo ELISPOT and multimer staining are well-established tests for the assessment of antigen-specific T cells. Many laboratories are now using a period of in vitro stimulation (IVS) to enhance detection. Here, we report the findings of a multi-centre panel organised by the Association for Cancer Immunotherapy Immunoguiding Program to investigate the impact of IVS protocols on the detection of antigen-specific T cells of varying ex vivo frequency. Five centres performed ELISPOT and multimer staining on centrally prepared PBMCs from 3 donors, both ex vivo and following IVS. A harmonised IVS protocol was designed based on the best-performing protocol(s), which was then evaluated in a second phase on 2 donors by 6 centres. All centres were able to reliably detect antigen-specific T cells of high/intermediate frequency both ex vivo (Phase I) and post-IVS (Phase I and II). The highest frequencies of antigen-specific T cells ex vivo were mirrored in the frequencies following IVS and in the detection rates. However, antigen-specific T cells of a low/undetectable frequency ex vivo were not reproducibly detected post-IVS. Harmonisation of the IVS protocol reduced the inter-laboratory variation observed for ELISPOT and multimer analyses by approximately 20?%. We further demonstrate that results from ELISPOT and multimer staining correlated after (P?R 2?=?0.5113), but not before IVS. In summary, IVS was shown to be a reproducible method that benefitted from method harmonisation.