Figure 3.

N-BPs-treated LNMSC drive Vδ2 T lymphocyte to effector memory T cells. (A and B) Peripheral blood T lymphocytes (A) or CFSE-labeled purified Vδ2 T cells (B) were co-cultured with LNMSC, either untreated or pre-treated for 12 h with Pam (5 μM) or Zol (1 μM), as indicated. On Day 10, percentage of Vδ2 T cells was evaluated by immunofluorescence using the anti-Vδ2 mAb PE-BB3 and FACS analysis (A). Mean±SD from 4 experiments. *P<0.001 versus T or Vδ2 cells cultured on untreated LNMSC. (B) Proliferation was assessed on the basis of the decreased green (CFSE^dull) fluorescence of dividing cells. Data were analyzed by the Modfit 3.1 computer program and expressed as cell number/generation of proliferating cells. (C and D) Immunophenotype of Vδ2 T cells cultured alone or co-cultured for 10 days with untreated or Pam or Zol pre-treated LNMSC, as indicated, stained with APC-anti-CD27 or PE-anti-CCR7 and FITC-anti-CD45RA. (C) Results are expressed as percentage of effector memory (EM) T cells, i.e. CD45RA-CD27- (white columns) or CD45RA-CCR7- cells (gray columns). Mean±SD from 6 experiments *P<0.001 versus Vδ2 T cells in the absence of LNMSC **P<0.001 versus Vδ2 T cells co-cultured with untreated LNMSC. (D) Vδ2 T cells alone (medium, left dot plots) or Vδ2 T cells co-cultured with untreated or treated LNMSC, as indicated, were stained as in (C). Results are expressed as log green fluorescence intensity (x-axis, MFI, a.u.) versus either log far red (first row) or red (second row) fluorescence intensity (y-axis, MFI, a.u.). One representative experiment out of 6 is shown. Percentages of positive cells in each quadrant show the naive (N, upper right, double positive), central memory (CM, upper left, CD45RA⁻ and CD27⁺ or CCR7⁺), effector memory (EM, lower left, double negative) and terminally differentiated effector memory (TEMRA, lower right, CD45RA⁺ but CD27⁻ or CCR7⁻) cells. Statistical analysis: two-tailed Student’s t-test.