Haematopoietic progenitor cells from the common marmoset as targets of gene transduction by retroviral and adenoviral vectors.
To establish a new non-human primate model for human cytokine and gene therapy, we characterized lymphocytes and haematopoietic progenitor cells of the small New World monkey, the common marmoset. We first assessed the reactions of marmoset bone marrow (BM) and peripheral blood (PB) cells to mouse anti-human monoclonal antibodies (mAbs) for the purpose of isolating marmoset lymphocytes and haematopoietic progenitor cells. Both cell fractions stained with CD4 and CD8 mAbs were identified as lymphocytes by cell proliferation assay and morphological examination. Myeloid-specific mAbs such as CD14 and CD33 did not react with marmoset BM and PB cells. No available CD34 and c-kit mAbs could be used to purify the marmoset haematopoietic progenitor cells. Furthermore, we studied the in vitro transduction of the bacterial beta-galactosidase (LacZ) gene into CFU-GM derived from marmoset BM using retroviral and adenoviral vectors. The transduction efficiency was increased by using a mixed culture system consisting of marmoset BM stromal cells and retroviral producer cells. It was also possible to transduce LacZ gene into marmoset haematopoietic progenitor cells with adenoviral vectors as well as retroviral vectors. The percentage of adenovirally transduced LacZ-positive clusters was 15% at day 4 (multiplicity of infection=200), but only 1-2% at day 14. The differential use of viral vector systems is to be recommended in targeting different diseases. Our results suggested that marmoset BM progenitor cells were available to examine the transduction efficiency of various viral vectors in vitro.
Hibino
H
H
Department of Hematology/Oncology, Institute of Medical Science, The University of Tokyo, Central Institute for Experimental Animals Laboratories Inc., Kawasaki, Kanagawa, Japan.
Tani
K
K
Sugiyama
H
H
Suzuki
S
S
Wu
M S
MS
Izawa
K
K
Hase
H
H
Nakazaki
Y
Y
Tanabe
T
T
Ooi
J
J
Izeki
T
T
Tojo
A
A
Saitoh
I
I
Tanioka
Y
Y
Asano
S
S
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology; D002144:Callithrix; D003429:Cross Reactions / Q000276:immunology; D005260:Female; D005822:Genetic Vectors / Q000592:standards; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007763:Lac Operon / Q000235:genetics; D008214:Lymphocytes / Q000276:immunology; D008297:Male; D023421:Models, Animal; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*
D000911:Antibodies, Monoclonal