Fig 4. Effect of anti-gal-9 mAbs on the “T_CM-like” phenotype shifting induced by gal-9 in resting PBMCs.

A-B. PBMCs freshly isolated from healthy donors were treated for one week with human recombinant gal-9 (Gal-9S; 40 nM) with or without combination with lactose (5 mM), control isotype antibody (ctrl IgG1), Gal-Nab1 or Gal-Nab2 (67 nM i.e. 10 μg/mL). A. Flow cytometry analysis was performed to determine the level of naive (CCR7⁺ CD45RO^-), central memory (CCR7⁺ CD45RO⁺) effector memory (CCR7^- CD45RO⁺) and effector (CCR7^- CD45RO^-) T cells within the CD3⁺ population. At least 50% of the cells were alive. Dead cells were gated out. B. For each treatment condition, percentages of cells with an apparent central memory phenotype were normalized with the basal level of this subpopulation in untreated cells. Data are represented as means ± SEM of four independent experiments with different donors. **p<0.01; ns: not significant; compared with gal-9 treatment (one-way ANOVA/Dunnet post-test).