Induction of γδ T-lymphocyte effector functions by bisphosphonate zoledronic acid in cancer patients in vivo by Francesco Dieli, Nicola Gebbia, Fabrizio.

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Induction of γδ T-lymphocyte effector functions by bisphosphonate zoledronic acid in cancer patients in vivo by Francesco Dieli, Nicola Gebbia, Fabrizio Poccia, Nadia Caccamo, Carla Montesano, Fabio Fulfaro, Carlo Arcara, Maria R. Valerio, Serena Meraviglia, Caterina Di Sano, Guido Sireci, and Alfredo Salerno Blood Volume 102(6):2310-2311 September 15, 2003 ©2003 by American Society of Hematology

Zoledronic acid activates in vitro Vγ9Vδ2 cells and induces subset distribution in cancer patients in vivo. Zoledronic acid activates in vitro Vγ9Vδ2 cells and induces subset distribution in cancer patients in vivo. (A) PBMCs were incubated in vitro with IPP (▵) or zoledronic acid (□) at the indicated final concentrations and 10 U/mL IL-2. Then, 7 days later, cells were collected and the percentage of Vγ9Vδ2+ cells within the CD3+ population was assessed. (B) Vγ9Vδ2 T-cell subsets were analyzed by 3-color fluorescence-activated cell-sorter (FACS) analysis before or after in vivo treatment with zoledronic acid. The following antibodies were used in different combinations: anti-Vδ2 fluorescein isothiocyanate (FITC, IMMU389; Coulter, Miami, FL), anti-CD27 phycoerythrin (PE) (M-T271; BD Pharmingen, San Diego, CA), and anti-CD45RA PE–cytochrome 5 (Cy5, 2H4; Coulter). Data were acquired on a FACSCalibur instrument (BD Biosciences, San Diego, CA) and analyzed using CellQuest software (BD Immunocytometry Systems, San Jose, CA). Error bars indicate standard deviation. Francesco Dieli et al. Blood 2003;102:2310-2311 ©2003 by American Society of Hematology

Proliferation and IFNγ production by Vγ9Vδ2 cells from cancer patients before and after treatment with zoledronic acid. Proliferation and IFNγ production by Vγ9Vδ2 cells from cancer patients before and after treatment with zoledronic acid. PBMCs were cultured at 37°C, in the presence of 5% CO2, at 106/mL in 96-well flat-bottomed plates (0.2 mL/well), with isopentenylpyrophosphate (IPP; Sigma Chemical, St Louis, MO; 100 μM/mL final concentration) and 20 U/mL final concentration human recombinant IL-2 (▪) or with IL-2 alone as a control (□). Proliferation (A) was measured 7 days later by adding 1 μCi/well (0.037 MBq) [3H]thymidine (Amersham, Arlington Heights, IL) during the last 6 hours of culture. Cells were then harvested and [3H]thymidine incorporation was measured with a liquid scintillation β-counter. Results are expressed as mean counts per minute (cpm) of triplicate wells ± standard deviation (SD). IFNγ levels in the 48-hour culture supernatants (B) were assessed by 2 monoclonal antibodies (mAbs) sandwich enzyme-linked immunosorbent assay (ELISA) assay following the manufacturer's recommendations (R&D Systems, Minneapolis, MN). Francesco Dieli et al. Blood 2003;102:2310-2311 ©2003 by American Society of Hematology