A fast and efficient HLA multimer-based sorting procedure that induces little apoptosis to isolate clinical grade human tumor specific T lymphocytes |
| |
Authors: | Régis Bouquié Annabelle Bonnin Karine Bernardeau Amir Khammari Brigitte Dréno Francine Jotereau Nathalie Labarrière François Lang |
| |
Institution: | (1) INSERM U892, 9 quai Moncousu, 44093 Nantes Cedex 1, France;(2) CHU of Nantes, Unit of Skin Cancer, 44093 Nantes, France;(3) Faculté des Sciences, Université de Nantes, 44322 Nantes, France;(4) UFR des Sciences Pharmaceutiques, Université de Nantes, 44322 Nantes, France |
| |
Abstract: | HLA multimers are now widely used to stain and sort CD8 T lymphocytes specific 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 first obstacle is
the use of streptavidin 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 multimers.
In the present report, we describe new HLA multimers composed of immunomagnetic beads covalently coupled to a mAb specific
for the AviTag peptide and coated with HLA/peptide monomers bearing the non biotinylated 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 multimers
with the previously described multimers made with streptavidin beads coated with biotinylated HLA/peptide monomers, in terms
of sorting efficiency, recovery of functional T cells, apoptosis and activation. We provide evidence that the new multimers
could very efficiently sort pure populations of T lymphocytes specific for three different melanoma antigens (Melan-A, gp100
and NA17-A) after a single peptide stimulation of melanoma patients’ PBMC. The recovered specific 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 specific T lymphocytes. Altogether,
these new multimers fulfill all the necessary requirements to select clinical grade T lymphocytes and should facilitate the
development of ACT protocols in cancer patients. |
| |
Keywords: | Immunotherapy Cell-sorting HLA multimers Melanoma T lymphocytes |
本文献已被 PubMed SpringerLink 等数据库收录! |
|