Abstract
Activation of cancer-specific T cells by tumor vaccines can lead to the rejection of the tumor. However, monitoring the tumor-specific T cell response continues to be a challenge. The magnitude and cytokine effector lineage of the vaccine-induced T cells, their ability to kill, and the avidity of the T cells for the tumor are among the primary parameters that define the T cell’s potential impact on the tumor. Due to determinant spreading, immunization with a tumor antigen can result in recruitment of T cells with specificity for unrelated antigens of the tumor. The efficacy of this second wave T cell repertoire will depend on clonal sizes, effector lineages, and their avidities for the tumor. Comprehensive immune monitoring should be able to measure all the above parameters of the primary and secondary T cell responses to the tumor. In this chapter, we propose an ELISPOT-based protocol which can help accomplish the above goal with reasonable cost and effort, requiring only 10 ml of patient blood.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Baars A, Claessen AM, van den Eertwegh AJ, Gall HE, Stam AG, Meijer S et al (2000) Skin tests predict survival after autologous tumor cell vaccination in metastatic melanoma: experience in 81 patients. Ann Oncol 11(8):965–970
Barth RJ Jr, Fisher DA, Wallace PK, Channon JY, Noelle RJ, Gui J et al (2010) A randomized trial of ex vivo CD40L activation of a dendritic cell vaccine in colorectal cancer patients: tumor-specific immune responses are associated with improved survival. Clin Cancer Res 16(22):5548–5556
Bartholomae WC, Rininsland FH, Eisenberg JC, Boehm BO, Lehmann PV, Tary-Lehmann M (2004) T cell immunity induced by live, necrotic, and apoptotic tumor cells. J Immunol 173(2):1012–1022
Bates GJ, Fox SB, Han C, Leek RD, Garcia JF, Harris AL et al (2006) Quantification of regulatory T cells enables the identification of high-risk breast cancer patients and those at risk of late relapse. J Clin Oncol 24(34):5373–5380
de Vries IJ, Bernsen MR, Lesterhuis WJ, Scharenborg NM, Strijk SP, Gerritsen MJ et al (2005) Immuno monitoring tumor-specific T cells in delayed-type hypersensitivity skin biopsies after dendritic cell vaccination correlates with clinical outcome. J Clin Oncol 23(24):5779–5787
Ding ZC, Zhou G (2012) Cytotoxic chemotherapy and CD4+ effector T cells: an emerging alliance for durable antitumor effects. Clin Dev Immunol 2012:890178
Disis ML (2011) Immunologic biomarkers as correlates of clinical response to cancer immunotherapy. Cancer Immunol Immunother 60(3):433–442
Disis ML, Schiffman K, Gooley TA, McNeel DG, Rinn K, Knutson KL (2000) Delayed-type hypersensitivity response is a predictor of peripheral blood T-cell immunity after HER-2/neu peptide immunization. Clin Cancer Res 6(4):1347–1350
Disis ML, Wallace DR, Gooley TA, Dang Y, Slota M, Lu H et al (2009) Concurrent trastuzumab and HER2/neu-specific vaccination in patients with metastatic breast cancer. J Clin Oncol 27(28):4685–4692
Dittrich M, Lehmann PV (2012) Statistical analysis of ELISPOT assays. Methods Mol Biol 792:173–183
Gulley JL, Arlen PM, Madan RA, Tsang KY, Pazdur MP, Skarupa L et al (2010) Immunologic and prognostic factors associated with overall survival employing a poxviral-based PSA vaccine in metastatic castrate-resistant prostate cancer. Cancer Immunol Immunother 59(5):663–674
Hardwick N, Chain B (2011) Epitope spreading contributes to effective immunotherapy in metastatic melanoma patients. Immunotherapy 3(6):731–733
Hawkins N, Self S, Wakefield J (2006) The automated counting of spots for the ELISpot assay. J Immunol Methods 316(1–2):52–58
Herzog BA, Ott PA, Dittrich MT, Quast S, Karulin AY, Kalbacher H et al (2004) Increased in vivo frequency of IA-2 peptide-reactive IFNgamma+/IL-4- T cells in type 1 diabetic subjects. J Autoimmun 23(1):45–54
Hesse MD, Karulin AY, Boehm BO, Lehmann PV, Tary-Lehmann M (2001) A T cell clone’s avidity is a function of its activation state. J Immunol 167(3):1353–1361
Hofstetter HH, Targoni OS, Karulin AY, Forsthuber TG, Tary-Lehmann M, Lehmann PV (2005) Does the frequency and avidity spectrum of the neuroantigen-specific T cells in the blood mirror the autoimmune process in the central nervous system of mice undergoing experimental allergic encephalomyelitis? J Immunol 174(8):4598–4605
Inderberg-Suso EM, Trachsel S, Lislerud K, Rasmussen AM, Gaudernack G (2012) Widespread CD4+ T-cell reactivity to novel hTERT epitopes following vaccination of cancer patients with a single hTERT peptide GV1001. Oncoimmunology 1(5):670–686
Jaffee EM, Hruban RH, Biedrzycki B, Laheru D, Schepers K, Sauter PR et al (2001) Novel allogeneic granulocyte-macrophage colony-stimulating factor-secreting tumor vaccine for pancreatic cancer: a phase I trial of safety and immune activation. J Clin Oncol 19(1):145–156
Karulin AY, Hesse MD, Tary-Lehmann M, Lehmann PV (2000) Single-cytokine-producing CD4 memory cells predominate in type 1 and type 2 immunity. J Immunol 164(4):1862–1872
Kenter GG, Welters MJ, Valentijn AR, Lowik MJ, Berends-van der Meer DM, Vloon AP, et al (2009) Vaccination against HPV-16 oncoproteins for vulvar intraepithelial neoplasia. N Engl J Med 361(19):1838–1847
Kirkwood JM, Lee S, Moschos SJ, Albertini MR, Michalak JC, Sander C et al (2009) Immunogenicity and antitumor effects of vaccination with peptide vaccine ± granulocyte-monocyte colony-stimulating factor and/or IFN-alpha2b in advanced metastatic melanoma: Eastern Cooperative Oncology Group Phase II Trial E1696. Clin Cancer Res 15(4):1443–1451
Kreher CR, Dittrich MT, Guerkov R, Boehm BO, Tary-Lehmann M (2003) CD4+ and CD8+ cells in cryopreserved human PBMC maintain full functionality in cytokine ELISPOT assays. J Immunol Methods 278(1–2):79–93
Kuerten S, Nowacki TM, Kleen TO, Asaad RJ, Lehmann PV, Tary-Lehmann M (2008) Dissociated production of perforin, granzyme B, and IFN-gamma by HIV-specific CD8(+) cells in HIV infection. AIDS Res Hum Retroviruses 24(1):62–71
Kuerten S, Batoulis H, Recks MS, Kraracsony E, Zhang W, Subbramanian RA et al (2012) Resting of cryopreserved PBMC does not in general benefit the performance of antigen-specific T cell ELISPOT assays. Cells 1(3):409–427
Lehmann PV, Zhang W (2012) Unique strengths of ELISPOT for T cell diagnostics. Methods Mol Biol 792:3–23
Lehmann PV, Forsthuber T, Miller A, Sercarz EE (1992) Spreading of T-cell autoimmunity to cryptic determinants of an autoantigen. Nature 358(6382):155–157
Lehmann PV, Sercarz EE, Forsthuber T, Dayan CM, Gammon G (1993) Determinant spreading and the dynamics of the autoimmune T-cell repertoire. Immunol Today 14(5):203–208
Lonsdorf AS, Kuekrek H, Stern BV, Boehm BO, Lehmann PV, Tary-Lehmann M (2003) Intratumor CpG-oligodeoxynucleotide injection induces protective antitumor T cell immunity. J Immunol 171(8):3941–3946
Lopez MN, Pereda C, Segal G, Munoz L, Aguilera R, Gonzalez FE et al (2009) Prolonged survival of dendritic cell-vaccinated melanoma patients correlates with tumor-specific delayed type IV hypersensitivity response and reduction of tumor growth factor beta-expressing T cells. J Clin Oncol 27(6):945–952
Mohan JF, Unanue ER (2012) Unconventional recognition of peptides by T cells and the implications for autoimmunity. Nat Rev Immunol 12(10):721–728
Mosmann TR, Kobie JJ, Lee FE, Quataert SA (2009) T helper cytokine patterns: defined subsets, random expression, and external modulation. Immunol Res 45(2–3):173–184
Nowacki TM, Kuerten S, Zhang W, Shive CL, Kreher CR, Boehm BO et al (2007) Granzyme B production distinguishes recently activated CD8(+) memory cells from resting memory cells. Cell Immunol 247(1):36–48
Quast S, Zhang W, Shive C, Kovalovski D, Ott PA, Herzog BA et al (2005) IL-2 absorption affects IFN-gamma and IL-5, but not IL-4 producing memory T cells in double color cytokine ELISPOT assays. Cell Immunol 237(1):28–36
Ramachandran H, Laux J, Moldovan I, Caspell P, Lehmann PV (2012) Optimal thawing of cryopreserved peripheral blood mononuclear cells for use in high-throughput human immune monitoring studies. Cells 1(3):313–324
Rininsland FH, Helms T, Asaad RJ, Boehm BO, Tary-Lehmann M (2000) Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity. J Immunol Methods 240(1–2):143–155
Schlingmann TR, Rininsland FH, Bartholomae WC, Kuekrek H, Lehmann PV, Tary-Lehmann M (2009a) Vaccination with tumor cells pulsed with foreign peptide induces immunity to the tumor itself. Clin Immunol 133(1):45–51
Schlingmann TR, Shive CL, Targoni OS, Tary-Lehmann M, Lehmann PV (2009b) Increased per cell IFN-gamma productivity indicates recent in vivo activation of T cells. Cell Immunol 258(2):131–137
Slingluff CL Jr, Petroni GR, Chianese-Bullock KA, Smolkin ME, Hibbitts S, Murphy C et al (2007) Immunologic and clinical outcomes of a randomized phase II trial of two multipeptide vaccines for melanoma in the adjuvant setting. Clin Cancer Res 13(21):6386–6395
Stern BV, Boehm BO, Tary-Lehmann M (2002) Vaccination with tumor peptide in CpG adjuvant protects via IFN-gamma-dependent CD4 cell immunity. J Immunol 168(12):6099–6105
Targoni OS, Lehmann PV (1998) Endogenous myelin basic protein inactivates the high avidity T cell repertoire. J Exp Med 187(12):2055–2063
Thakur A, Pedersen LE, Jungersen G (2012) Immune markers and correlates of protection for vaccine induced immune responses. Vaccine 30(33):4907–4920
Tian J, Lehmann PV, Kaufman DL (1997) Determinant spreading of T helper cell 2 (Th2) responses to pancreatic islet autoantigens. J Exp Med 186(12):2039–2043
Tigno-Aranjuez JT, Jaini R, Tuohy VK, Lehmann PV, Tary-Lehmann M (2009a) Encephalitogenicity of complete Freund’s adjuvant relative to CpG is linked to induction of Th17 cells. J Immunol 183(9):5654–5661
Tigno-Aranjuez JT, Lehmann PV, Tary-Lehmann M (2009b) Dissociated induction of cytotoxicity and DTH by CFA and CpG. J Immunother 32(4):389–398
Zhang W, Lehmann PV (2012) Objective, user-independent ELISPOT data analysis based on scientifically validated principles. Methods Mol Biol 792:155–171
Zhang W, Caspell R, Karulin AY, Ahmad M, Haicheur N, Abdelsalam A et al (2009) ELISPOT assays provide reproducible results among different laboratories for T-cell immune monitoring–even in hands of ELISPOT-inexperienced investigators. J Immunotoxicol 6(4):227–234
Zhang W, Moldovan I, Targoni OS, Subbramanian RA, Lehmann PV (2012) How much of virus-specific CD8 T cell reactivity is detected with a peptide pool when compared to individual peptides? Viruses 4(11):2636–2649
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Lehmann, P.V., Sundararaman, S. (2013). When Results of T cell Immune Monitoring Match/Do Not Match Clinical Outcomes of Tumor Vaccine Trials: What More Could and Should We Measure?. In: Shurin, M., Umansky, V., Malyguine, A. (eds) The Tumor Immunoenvironment. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-6217-6_32
Download citation
DOI: https://doi.org/10.1007/978-94-007-6217-6_32
Published:
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-007-6216-9
Online ISBN: 978-94-007-6217-6
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)