Pe conjugated human cd45 antibody clone hi30

Primary human T-ALL samples were obtained from the Barts Hospital (London, UK) after informed consent via a protocol approved by the East London Research Ethics Committee and carried out in accordance with the principles of the Helsinki declaration (see Supplementary Table 2 for details), prior to treatment being administered to the patients. Primary cells from two distinct patients were immunophenotyped, and CD45⁺/CD7⁺/CD4^-/low/CD8^-/low cells sorted and infused I.V. in a non-conditioned Osterix-CreGFP/NOD/SCID/γ recipient mice. Primary xenograft transplantation was assessed via peripheral blood sampling and/or bone marrow aspiration. Bone marrow and spleen-derived primary xenografts were infused I.V. in non-conditioned NOD/SCID/γ secondary recipient mice for therapy experiments. Intravital imaging was performed as described above. Human T-ALL cells were labelled by injecting 10 μg of PE-conjugated human CD45 antibody (clone HI30, Biolegend) 15-30 minutes prior to the imaging session. For dexamethasone therapy experiments, mice were treated with daily injections of 15mg/kg I.V31 (link). Number of human T-ALL cells in therapy experiments was quantified using reference beads as described in36 (link).