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Fit for purpose? A case study: validation of immunological endpoint assays for the detection of cellular and humoral responses to anti-tumour DNA fusion vaccines

  • Focussed Research Review
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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Clinical trials are governed by an increasingly stringent regulatory framework, which applies to all levels of trial conduct. Study critical immunological endpoints, which define success or failure in early phase clinical immunological trials, require formal pre-trial validation. In this case study, we describe the assay validation process, during which the sensitivity, and precision of immunological endpoint assays were defined. The purpose was the evaluation of two multicentre phase I/II clinical trials from our unit in Southampton, UK, which assess the effects of DNA fusion vaccines on immune responses in HLA-A2+ patients with carcinoembryonic antigen (CEA)-expressing malignancies and prostate cancer. Validated immunomonitoring is being performed using ELISA and IFNγ ELISPOTs to assess humoral and cellular responses to the vaccines over time. The validated primary endpoint assay, a peptide-specific CD8+ IFNγ ELISPOT, was tested in a pre-trial study and found to be suitable for the detection of low frequency naturally occurring CEA- and prostate-derived tumour-antigen-specific T cells in patients with CEA-expressing malignancies and prostate cancer.

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Acknowledgments

Serum samples from healthy volunteers vaccinated with tetanus toxoid were kindly provided by Dr. Gianfranco Di Genova. We would like to thank Dr. Richard Sugar (QA Manager, Drug Development Office, Cancer Research UK) for his valuable support and assistance in the planning and documentation of the assay validations. Our thanks to Dr. J.P.Kerr for collation of clinical data for the study of pre-existing cellular immunity. AM and CO are supported by Cancer Research UK (CR-UK), FC is supported by Experimental Cancer Medicine Centre (ECMC) grant funding (joint CR–UK and the Department of Health of England, Scotland, Wales and Northern Ireland).

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None declared.

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Authors and Affiliations

  1. Cancer Sciences Division, School of Medicine, Southampton General Hospital, University of Southampton, Somers Cancer Research Building, Mailpoint 824, Tremona Road, Southampton, SO16 6YD, UK

    Ann Mander, Ferdousi Chowdhury, Lindsey Low & Christian H. Ottensmeier

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  1. Ann Mander
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  2. Ferdousi Chowdhury
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  3. Lindsey Low
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  4. Christian H. Ottensmeier
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Correspondence to Ann Mander.

Additional information

This paper is a Focussed Research Review based on a presentation given at the Sixth Annual Meeting of the Association for Immunotherapy of Cancer (CIMT), held in Mainz, Germany, 15–16 May 2008.

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Mander, A., Chowdhury, F., Low, L. et al. Fit for purpose? A case study: validation of immunological endpoint assays for the detection of cellular and humoral responses to anti-tumour DNA fusion vaccines. Cancer Immunol Immunother 58, 789–800 (2009). https://doi.org/10.1007/s00262-008-0633-z

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