[Efficient gene transfer mediated by retrovirus vector in leukemic cells].
OBJECTIVE
To establish an efficient and safe gene transfer system mediated by retrovirus for providing an experimental basis of gene therapy in human leukemia.


METHODS
The retrovirus vector pLXSN, carrying the NeoR, was transferred into amphotropic packaging cell line GP + envAm12 by liposome method or by transduction. Retrovirus with higher titer was used to infect human leukemic cell lines NB4, U937 and THP-1. The efficiency of gene transduction was tesed on colonies formed by K562 cells after gene transduction.


RESULTS
The titer of DOSPER liposome transfected GP + envAm12 pool on NIH3T3 cells was 8.0 x 10(5) CFU/ml while that of producer pool infected with mouse retrovirus was 1.6 x 10(7) CFU/ml. Integration of NeoR into human leukemic cells was confirmed by PCR. By both nested PCR and NeoR rescue assay, no helper virus was found. NeoR gene transfer with the efficiency as high as 93.3 to 100% in K562 cells was verified by semi-nested PCR on the colonies after seven days' culture.


CONCLUSIONS
The efficiency and safety of the gene transfer system may provide an optimal experimental system in gene therapy for human leukemia.
Jiangsu Institute of Hematology, First Affiliated Hospital of Suzhou Medical College.
J
J
Fu
Z
Z
Chen
J
J
Cen
D000818:Animals; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D020014:K562 Cells; D007938:Leukemia / Q000473:pathology / Q000628:therapy*; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods; D012190:Retroviridae / Q000235:genetics*; D014407:Tumor Cells, Cultured