Long-term high-level reconstitution of NADPH oxidase activity in murine X-linked chronic granulomatous disease using a bicistronic vector expressing gp91phox and a ΔLNGFR cell surface marker

Mohammed A. Sadat, Nancy Pech, So Saulnier, Brendan A. Leroy, Johann P. Hossle, Manuel Grez, Mary C. Dinauer

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

A murine model of X-linked chronic granuiomatous disease (X-CGD), an inherited immune deficiency with absent phagocyte NADPH oxidase activity caused by defects in the gp91phox gene, was used to evaluate a bicistronic retrovirai vector in which expression of human gp91phox and a linked gene for ΔLNGFR, a truncated form of human low-affinity nerve growth factor receptor, are under the control of a spleen focus-forming virus long-terminal repeat (LTR). Four independent cohorts of 11-Gy irradiated X-CGD mice (total, 22 mice) were transplanted with or without preselection of transduced X-CGD bone marrow (BM). Transplanted mice had high-level correction of neutrophii gp91phox expression and reconstitution of NADPH oxidase activity. Expression lasted for at least 14 months in primary transplants, and persisted in secondary and tertiary transplants. Both gp91phox and ΔLNGFR were detected on circulating granulocytes, iymphocytes, lymphoid, and (for ΔLNGFR) red blood cells. Mice receiving transduced bone marrow [BM] preselected ex vivo for ΔLNGFR expression had high-level (= 80%) reconstitution with transduced cells, with an improved fraction of oxidase-corrected neutrophils posttransplant. Analysis of secondary and tertiary CFU-S showed that silencing of individual provirus integrants can occur even after preselection for ΔLNGFR prior to transplantation, and that persistent provirus expression was associated with multiple integration sites in most cases. No obvious adverse consequences of transgenic protein expression were observed.

Original languageEnglish
Pages (from-to)651-666
Number of pages16
JournalHuman Gene Therapy
Volume14
Issue number7
DOIs
StatePublished - May 1 2003

Fingerprint

Dive into the research topics of 'Long-term high-level reconstitution of NADPH oxidase activity in murine X-linked chronic granulomatous disease using a bicistronic vector expressing gp91phox and a ΔLNGFR cell surface marker'. Together they form a unique fingerprint.

Cite this