Analysis of gene transfer efficiency of retrovirus producer cell transplantation for in situ gene transfer to hematopoietic cells.
OBJECTIVE
The aim of this study was to assess the gene transfer efficiency of an in situ administration protocol for hematopoietic stem/progenitor cells in the rhesus macaque (Macaca mulatta) animal model.


METHODS
Moloney murine leukemia virus amphotropic vector producer cells (1--2 x 10(8) cells/animal) were transplanted into the femoral bone marrow cavities of six macaques. To determine if the levels of gene transfer could be increased, a second injection at the same dose of producer cells was performed into the iliac crest in three of the six macaques.


RESULTS
We demonstrated that 0.02-0.1% of peripheral blood mononuclear cells contained the vector transgene for up to 12 months following the initial administration of producer cells. Hematopoietic progenitor cell assays indicated that the neomycin phosphotransferase gene was detected in 10--30% of progenitor cell colonies. A humoral immune response directed toward viral particles was demonstrated in all animals. Additionally, we demonstrated that an increase in the levels of transduced cells, up to 1% of circulating peripheral blood mononuclear cells and granulocytes, contain the transgene following producer cell readministration.


CONCLUSIONS
These data demonstrate the successful in situ gene transfer to hematopoietic stem/progenitor cells and circulating peripheral blood mononuclear cells that persists as long as 12 months postinjection, in the absence of any preconditioning.
Newbound
G C
GC
Children's Research Institute, Children's Hospital, Columbus, OH 43205, USA.
Cooper
J R
JR
O'Rourke
J P
JP
Baskin
C R
CR
Bunnell
B A
BA
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D001853:Bone Marrow; D002460:Cell Line; D017690:Cell Transplantation; D005269:Femur; D005434:Flow Cytometry; D015870:Gene Expression; D022761:Gene Transfer, Horizontal; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000378:metabolism*; D019868:Kanamycin Kinase / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D008253:Macaca mulatta; D008979:Moloney murine leukemia virus / Q000235:genetics* / Q000276:immunology; D012313:RNA / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000914:Antibodies, Viral; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D012313:RNA; D019868:Kanamycin Kinase