Page 166 - Haematologica April 2020

V.P. Vaikari et al.
EV and CD99-S tissues compared with CD99-L tissues, confirming a decrease in leukemia engraftment in mice with CD99-L cells (Figure 6I-M).
In a primary blast murine model, we engrafted primary AML samples (AML-4) transduced with CD99-L (n=3) or EV (n=3) in sub-lethally irradiated mice. Mice were sac-
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rificed four months post engraftment (Figure 6N-O). CD99-L mice had less BM engraftment, though this was not significant compared with the EV mice (1.52 vs. 4.76%, P=0.1) (Figure 6P), and a significant decrease in hCD45+ cells in PB (7.5% vs. 26.9 %, P=0.005) (Figure 6Q and Online Supplementary Figures S10 and S12).
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Figure 7. Effect of CD99 mAB on acute myeloid leukemia (AML) cells. (A) Cell viability of AML blasts treated with 20μg/mL of mAbCD99 for 48 hours (h) and meas- ured using trypan-blue (n=7). (B) Total number of colonies comparison between AML blasts treated with mAb CD99 and control blasts (n=6). (C) Viability of THP-1, MOLM-13 and healthy donor peripheral blood mononuclear cells (PBMC) treated with 5μg/mL of mAbCD99 for 48 h and measured using alamar blue. (D and E) Apoptosis measured in THP-1 and MOLM-13 cells treated with mAbCD99 for 72 h and stained with Annexin V for flow cytometry analysis. (F and G) Quantitative analy- sis and representative images of migration of THP-1, MOLM-13 and PBMC treated with mAB CD99 towards SDF-1α performed in a transwell plate. (H) CD11b meas- ured by flow cytometry in THP-1 cells 72 h post treatment with 2.5 μg/mL of mAB CD99. (I) Representative images for Wright-Giemsa staining of THP-1 cells treated with 2.5 μg/mL of mAb CD99 for 72 h. (***P<0.001; **P<0.01; *P<0.05).
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haematologica | 2020; 105(4)