An in vitro-transcribed-mRNA polyepitope construct encoding 32 distinct HLA class I-restricted epitopes from CMV, EBV, and Influenza for use as a functional control in human immune monitoring studies

Journal of Immunological Methods
Julie S NielsenJohn R Webb

Abstract

Interest and activity in the areas of clinical immunotherapy and therapeutic vaccines are growing dramatically, thus there is a pressing need to develop robust tools for assessment of vaccine-induced immunity. CD8+ T cell immunity against specific antigens is normally measured by either flow cytometry using MHC tetramer reagents or via biological assays such as intracellular cytokine staining or ELISPOT after stimulation with specific peptide epitopes. However, these methodologies depend on precise knowledge of HLA-restricted epitopes combined with HLA typing of subjects. As an alternative approach, electroporation of antigen presenting cells (APC) with in vitro-transcribed mRNA (IVT-mRNA) encoding the antigen of interest bypasses the requirements for HLA typing and knowledge of specific epitopes. A current limitation of the IVT-mRNA technique is the lack of robust positive control RNAs to verify the efficacy of electroporation and to ensure that the electroporated APC retain the ability to stimulate T cells. Herein we describe an IVT-mRNA construct wherein all 32 HLA class I-restricted epitopes of the widely used CEF (Cytomegalovirus, Epstein-Barr Virus and Influenza Virus) positive control peptide pool have been genetically s...Continue Reading

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Citations

Jul 26, 2011·Journal of Immunological Methods·Guang Lan ZhangVladimir Brusic
Sep 16, 2015·Annals of Oncology : Official Journal of the European Society for Medical Oncology·S D MartinB H Nelson
Nov 23, 2016·European Journal of Immunology·Mélissa MathieuNathalie Labrecque
Aug 14, 2013·Infection and Immunity·Xianqiong ZouMark C Herzberg
Dec 31, 2014·The Journal of Immunology : Official Journal of the American Association of Immunologists·Colleen R ZaccardRobbie B Mailliard
Dec 4, 2015·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Julie S NielsenBrad H Nelson
Jun 6, 2021·Briefings in Functional Genomics·Xinhui CaiJinyan Li

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