Extended Data Fig. 4: Flow cytometry analyses of RBCs in SCD patients.
From: Long-term outcomes of lentiviral gene therapy for the β-hemoglobinopathies: the HGB-205 trial
(a) Controls showing that the Pacific Blue (PB) labeled anti-HbA monoclonal antibody (Rockland) does not cross react with HbS in individualized RBCs by flow cytometry analysis. S/S RBC exhibit low level of fluorescence because the anti-HbA antibody also reacts with HbA2 (data from Rockland). However, the anti-HbA antibody does not cross react with HbF (data from Rockland). Because the epitope recognized by the anti-HbA antibody is also present on HbAT87Q, the latter is equally well recognized (data not shown). (b) Distribution of γ-globin and βA-T87Q-globin expressing RBCs in SCD patients’ blood by flow cytometry using antibodies that recognize either βA-T87Q-globin (anti-HbA PB-labeled) or γ-globin (anti-HbF PE-labeled). Co-stainings performed at M36, M14 and M15 for SCD-1, SCD-2 and SCD-3, respectively, showing reduced levels of HbF in high containing HbAT87Q RBCs, and inversely, for SCD-2 and SCD-3. (c) Distribution of βA-T87Q-globin vs. γ-globin expressing RBCs in SCD patients’ blood by flow cytometry using antibodies that recognize either βA-T87Q-globin (left) or γ-globin (right). M, months after GT; PB, Pacific Blue; PE, phycoerythrin. Corresponding datasets are represented in Tables 2a and b.
