Simultaneous assessment of cytotoxic T lymphocyte responses against multiple viral infections by combined usage of optim
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BioMed Central
Open Access
Methodology
Simultaneous assessment of cytotoxic T lymphocyte responses against multiple viral infections by combined usage of optimal epitope matrices, anti- CD3 mAb T-cell expansion and "RecycleSpot" Florian K Bihl1, Elisabetta Loggi3, John V Chisholm III1, Hannah S Hewitt1, Leah M Henry1, Caitlyn Linde1, Todd J Suscovich1, Johnson T Wong2, Nicole Frahm1, Pietro Andreone3 and Christian Brander*1 Address: 1Partners AIDS Research Center, Massachusetts General Hospital, Harvard Medical School, Boston, USA, 2Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, USA and 3Dipartimento di Cardioangiologia ed Epatologia, Ospedale S. OrsolaMalpighi, Università degli Studi di Bologna, Italy Email: Florian K Bihl - [email protected]; Elisabetta Loggi - [email protected]; John V Chisholm - [email protected]; Hannah S Hewitt - [email protected]; Leah M Henry - [email protected]; Caitlyn Linde - [email protected]; Todd J Suscovich - [email protected]; Johnson T Wong - [email protected]; Nicole Frahm - [email protected]; Pietro Andreone - [email protected]; Christian Brander* - [email protected] * Corresponding author
Published: 11 May 2005 Journal of Translational Medicine 2005, 3:20
doi:10.1186/1479-5876-3-20
Received: 09 March 2005 Accepted: 11 May 2005
This article is available from: http://www.translational-medicine.com/content/3/1/20 © 2005 Bihl et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Cytotoxic T CellsHIVEBVCMVHCVHBVCTLepitopepeptidecell expansionanti-CD3ELISpotpeptide matrix
Abstract The assessment of cellular anti-viral immunity is often hampered by the limited availability of adequate samples, especially when attempting simultaneous, high-resolution determination of T cell responses against multiple viral infections. Thus, the development of assay systems, which optimize cell usage, while still allowing for the detailed determination of breadth and magnitude of virusspecific cytotoxic T lymphocyte (CTL) responses, is urgently needed. This study provides an upto-date listing of currently known, well-defined viral CTL epitopes for HIV, EBV, CMV, HCV and HBV and describes an approach that overcomes some of the above limitations through the use of peptide matrices of optimally defined viral CTL epitopes in combination with anti-CD3 in vitro T cell expansion and re-use of cells from negative ELISpot wells. The data show that, when compared to direct ex vivo cell preparations, antigen-unspecific in vitro T cell expansion maintains the breadth of detectable T cell responses and demonstrates that harvesting cells from negative ELISpot wells for re-use in subsequent ELISpot assays (RecycleSpot), further maximized the use of available cells. Further
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