Skip to Main content Skip to Navigation
New interface
Journal articles

A fast and efficient HLA multimer-based sorting procedure that induces little apoptosis to isolate clinical grade human tumor specific T lymphocytes

Abstract : HLA multimers are now widely used to stain and sort CD8 T lymphocytes speciWc for epitopes from viral or tumoral antigens presented in an HLA class I context. However, the transfer of this technology to a clinical setting to obtain clinical grade CD8 T lymphocytes that may be used in adoptive cell transfer (ACT) is hindered by two main obstacles: the Wrst obstacle is the use of streptavi-din or derived products that are not available in clinical grade to multimerize HLA/peptide monomers and the second is the reported high degree of apoptosis that eventually occurs when T cell receptors are crosslinked by HLA multi-mers. In the present report, we describe new HLA multi-mers composed of immunomagnetic beads covalently coupled to a mAb speciWc for the AviTag peptide and coated with HLA/peptide monomers bearing the non bio-tinylated AviTag at the COOH terminus of the HLA heavy chain. Thus, all the components of this new reagent can be obtained in clinical grade. We compared these new multi-mers with the previously described multimers made with streptavidin beads coated with biotinylated HLA/peptide monomers, in terms of sorting eYciency, recovery of functional T cells, apoptosis and activation. We provide evidence that the new multimers could very eYciently sort pure populations of T lymphocytes speciWc for three diVer-ent melanoma antigens (Melan-A, gp100 and NA17-A) after a single peptide stimulation of melanoma patients' PBMC. The recovered speciWc T cells were cytotoxic against the relevant melanoma cell-lines and, in most cases, produced cytokines. In addition, in marked contrast with streptavidin-based multimers, our new multimers induced very little apoptosis or activation after binding speciWc T lymphocytes. Altogether, these new multimers fulWll all the necessary requirements to select clinical grade T lympho-cytes and should facilitate the development of ACT protocols in cancer patients.
Complete list of metadata

Cited literature [33 references]  Display  Hide  Download
Contributor : Nathalie LABARRIERE Connect in order to contact the contributor
Submitted on : Monday, February 17, 2020 - 5:35:39 PM
Last modification on : Thursday, September 1, 2022 - 11:10:29 AM
Long-term archiving on: : Monday, May 18, 2020 - 6:34:05 PM


 Restricted access
To satisfy the distribution rights of the publisher, the document is embargoed until : jamais

Please log in to resquest access to the document



Régis Bouquié, Annabelle Bonnin, Karine Bernardeau, Amir Khammari, Brigitte Dréno, et al.. A fast and efficient HLA multimer-based sorting procedure that induces little apoptosis to isolate clinical grade human tumor specific T lymphocytes. Cancer Immunology, Immunotherapy, 2008, 58 (4), pp.553-566. ⟨10.1007/s00262-008-0578-2⟩. ⟨inserm-02481983⟩



Record views