Cancer Immunology, Immunotherapy

, Volume 57, Issue 2, pp 185–195

PBMC are as good a source of tumor-reactive T lymphocytes as TIL after selection by Melan-A/A2 multimer immunomagnetic sorting


    • INSERM U601
  • Nadine Gervois
    • INSERM U601
    • Faculté des SciencesUniversité de Nantes
  • Annabelle Bonnin
    • INSERM U601
  • Régis Bouquié
    • INSERM U601
    • INSERM U601
    • Faculté des SciencesUniversité de Nantes
  • François Lang
    • INSERM U601
    • UFR des Sciences PharmaceutiquesUniversité de Nantes
Original Article

DOI: 10.1007/s00262-007-0361-9

Cite this article as:
Labarrière, N., Gervois, N., Bonnin, A. et al. Cancer Immunol Immunother (2008) 57: 185. doi:10.1007/s00262-007-0361-9


Choosing a reliable source of tumor-specific T lymphocytes and an efficient method to isolate these cells still remains a critical issue in adoptive cellular therapy (ACT). In this study, we assessed the capacity of MHC/peptide based immunomagnetic sorting followed by polyclonal T cell expansion to derive pure polyclonal and tumor-reactive Melan-A specific T cell populations from melanoma patient’s PBMC and TIL. We first demonstrated that this approach was extremely efficient and reproducible. We then used this procedure to compare PBMC and TIL-derived cells from three melanoma patients in terms of avidity for Melan-A A27L analog, Melan-A26–35 and Melan-A27–35, tumor reactivity (lysis and cytokine production) and repertoire. Regardless of their origin, i.e., fresh PBMC, peptide stimulated PBMC or TIL, all sorted populations (from the three patients) were cytotoxic against HLA-A2+ melanoma cell lines expressing Melan-A. Although some variability in peptide avidity, lytic activity and cytokine production was observed between populations of different origins in a given patient, it differed from one patient to another and thus no correlation could be drawn between T cell source and reactivity. Analysis of Vβ usage within the sorted populations showed the recurrence of Vβ3 and Vβ14 subfamilies in the three patients but differences in the rest of the Melan-A repertoire. In addition, in two patients, we observed major repertoire differences between populations sorted from the three sources. We especially documented that in vitro peptide stimulation of PBMC, used to facilitate the sort by enriching in specific T lymphocytes, could significantly alter their repertoire and reactivity towards tumor cells. We conclude that PBMC which are easily obtained from all melanoma patients, can be as good a source as TIL to derive high amounts of tumor-reactive Melan-A specific T cells, with this selection/amplification procedure. However, the conditions of peptide stimulation should be improved to prevent a possible loss of reactive clonotypes.


Melan-AMHC/peptideImmunomagnetic sortingMelanomaAdoptive immunotherapy

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© Springer-Verlag 2007