Polyclonal long-term MFGS-gp91phox marking in rhesus macaques after nonmyeloablative transplantation with transduced autologous peripheral blood progenitor cells.
We have recently reported that the RD114-pseudotyped MFGS-gp91phox vector achieves unprecedented levels of correction of the NADPH-oxidase gp91phox (approved gene symbol CYBB) defect in CD34(+) cells from patients with X-linked chronic granulomatous disease in the NOD/SCID mouse model. Considering clinical use of this vector, we transplanted autologous mobilized peripheral blood CD34(+) progenitor cells, transduced with the RD114-MFGS-gp91phox vector, into two healthy rhesus macaques following nonmyeloablative conditioning. The moderately high levels of in vivo marking seen in the first months following transduction decreased and stabilized at about 8 months posttransplant. Marking for both healthy animals after 15 months was 0.3 to 1.3 vector copies per 100 cells in lymphocytes, neutrophils, and monocytes. Vector insertion analyses performed by linear amplification-mediated PCR and sequencing identified 32 and 45 separate insertion sites in the animals. Identical insertion sites were found in myeloid cells and lymphocytes, demonstrating the successful transduction of lymphomyeloid progenitors. Some inserts landed in the vicinity of genes controlling cell cycle and proliferation. Statistical analyses of insertion sites 1 year posttransplant suggest a high diversity of insertion sites despite low marking.
Brenner
Sebastian
S
Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA. sebastian.brenner@uniklinikum-dresden.de
Ryser
Martin F
MF
Choi
Uimook
U
Whiting-Theobald
Narda
N
Kuhlisch
Eberhard
E
Linton
Gilda
G
Kang
Elizabeth
E
Lehmann
Romy
R
Rosen-Wolff
Angela
A
Rudikoff
Andrew G
AG
Farese
Ann M
AM
Macvittie
Thomas J
TJ
Roesler
Joachim
J
Horwitz
Mitchell E
ME
Malech
Harry L
HL
D000818:Animals; D018952:Antigens, CD34 / Q000032:analysis; D017988:Gammaretrovirus / Q000235:genetics; D018628:Gene Dosage; D005822:Genetic Vectors; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells; D008253:Macaca mulatta; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics*; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000235:genetics*; D014161:Transduction, Genetic; D019076:Transgenes; D019172:Transplantation Conditioning; D014182:Transplantation, Autologous; D016662:Virus Integration; D014916:Whole-Body Irradiation
D018952:Antigens, CD34; D008562:Membrane Glycoproteins; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases