Analysis of expression of hG6PD in hematopoietic cells of reconstituted mice.

(A) Analysis of hG6PD expression by cellulose acetate gel electrophoresis; h indicates human G6PD homodimer; m, mouse G6PD homodimer; hm, human-mouse heterodimer. C, lysate from an untransplanted control mouse. Mouse 5 illustrates loss of expression 4 months after BMT. Mouse 11 illustrates stable long-term expression in RBCs (R), WBCs (W), spleen (S), BM (B), and thymus (T). Mouse S5 was analyzed 14 months after receiving a transplant from a primary recipient that served as donor 11 months after the primary BMT. (B) Similar levels of hG6PD expression in RBCs and WBCs in individual transplanted mice. For both cell types hG6PD activity was normalized to that of the endogenous mouse G6PD. The slope of the regression line (y = 1.2 × −0.3) is very close to 1. (C) Expression of hG6PD doubles the overall G6PD level in peripheral blood RBCs. Average and SD values are shown for untransplanted control mice (B6) (n = 8) and for long-term expressing primary (I) BMT recipients (n = 16) and secondary (II) BMT recipients (n = 14). (D) Survival of hG6PD expression (Kaplan-Meyer method): VSV-G pseudotyped versus ecotropic virus. Each individual mouse still expressing hG6PD is shown by a short vertical line above each survival curve.