Gene transfer by MPIN into PNH-phenotype primary erythroid cells from a patient with PNH restores PIG-A function and resistance to hemolysis.

(A) GPI-AP (CD59), NGFR, E6, and CD45 surface expression of MPIN-transduced erythroid cells generated from GPI-AP⁻(CD59⁻) BMMCs on day 12. (B) Detection of the region including PIG-A in MPIN-transduced BMMCs by PCR. DNA was isolated from transduced (MN or MPIN) or untransduced (mock) BMMCs. A region including vector plus 5′ PIG-A was amplified by PCR. For positive control, the MPIN vector plasmid was used as a template, and Marker 6 was loaded as a marker. (C) (D) Small-scaled Ham test (panel C), and aerolysin assay (panel D). The percentage of lysis of CD59⁺NGFR⁺ (+ +) and CD59⁻NGFR⁻ (− −) cells from MPIN-transduced BMMCs were compared. For 100% lysis, water was used in place of serum, and for 0% lysis, heat-inactivated serum was used. Results are shown as mean ± SE.