Efficient transduction and selection of human T-lymphocytes with bicistronic Thy1/HSV1-TK retroviral vector produced by a human packaging cell line

2004 ◽  
Vol 6 (4) ◽  
pp. 374-386 ◽  
Author(s):  
François M. Lemoine ◽  
Mariana Mesel-Lemoine ◽  
Mustapha Cherai ◽  
Géraldine Gallot ◽  
Henri Vié ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Cell Line ◽  
Retroviral Vector ◽  
Packaging Cell Line ◽  
Packaging Cell ◽  
Human T Lymphocytes ◽  
Selection Of

scholarly journals Defective Retroviral Endogenous RNA Is Efficiently Transmitted by Infectious Particles Produced on an Avian Retroviral Vector Packaging Cell Line

Virology ◽  
1995 ◽  
Vol 207 (1) ◽  
pp. 271-275 ◽  
Author(s):  
Corinne Ronfort ◽  
Anne Girod ◽  
François-Loı̈c Cosset ◽  
Catherine Legras ◽  
Victor Marc Nigon ◽  
...  
Keyword(s):  
Cell Line ◽  
Retroviral Vector ◽  
Packaging Cell Line ◽  
Packaging Cell

scholarly journals Association of proliferating cell nuclear antigen with cyclin-dependent kinases and cyclins in normal and transformed human T lymphocytes

Blood ◽  
1994 ◽  
Vol 84 (10) ◽  
pp. 3413-3421 ◽  
Author(s):  
A Szepesi ◽  
EW Gelfand ◽  
JJ Lucas
Keyword(s):  
T Cells ◽  
T Lymphocytes ◽  
Cell Line ◽  
Proliferating Cell Nuclear Antigen ◽  
Kinase Activity ◽  
Jurkat Cells ◽  
Nuclear Antigen ◽  
Human T Lymphocytes ◽  
Normal Human ◽  
Proliferating Cell

Abstract The proliferating cell nuclear antigen (PCNA) is an auxiliary protein of DNA polymerase delta and appears to be needed for both DNA synthesis and DNA repair. It is present in low amount in resting normal human T lymphocytes and, upon mitogenic stimulation with phorbol dibutyrate and ionomycin, begins to increase in mid-G1 phase, approximately 12 to 15 hours before entry into S phase. PCNA continues to increase in amount throughout the cell cycle and remains high in proliferating cultures. PCNA was extracted from activated normal T cells and from the transformed T-lymphoblastoid cell line Jurkat by a method that recovered approximately 98% of total cellular PCNA but yet retained its associations with other proteins. PCNA immunoprecipitates possessed H1 histone kinase activity, which increased in parallel with increasing cellular content of PCNA. Both the cdc2 and cdk2 kinases were found associated with PCNA in normal T cells, in amounts consistent with detected kinase activity. The results indicate that PCNA is not an inhibitory molecule of cdk/cyclin activity. Both normal and transformed T cells contained PCNA in association with cdk2, cdk4, cdk5, and cdk6, with the amount of PCNA associated with these molecules increasing in the order listed. Relatively high amounts of PCNA were also found associated with cyclins D2 and D3, the major cyclin partners of cdk6 in T cells. Though detected in normal cells, PCNA/cdc2 complexes were present in exceedingly low amount, if at all, in Jurkat cells. This cell line appeared to contain more of nearly all of the cdk and cyclin molecules analyzed, but there seemed to be little difference in the patterns of association of these molecules with PCNA in the cell line as compared with normal human T cells.

Development and testing of a packaging cell line for avian retroviral vectors

1991 ◽  
Vol 119 (3-4) ◽  
pp. 257-264 ◽  
Author(s):  
N. L. Meyers ◽  
S. C. Booth ◽  
N. Bumstead ◽  
L. Vick ◽  
K. Simkiss
Keyword(s):  
Cell Line ◽  
Retroviral Vectors ◽  
Packaging Cell Line ◽  
Packaging Cell

Highly efficient gene transfer into baboon marrow repopulating cells using GALV-pseudotype oncoretroviral vectors produced by human packaging cells

Blood ◽  
2002 ◽  
Vol 100 (12) ◽  
pp. 3960-3967 ◽  
Author(s):  
Peter A. Horn ◽  
Max S. Topp ◽  
Julia C. Morris ◽  
Stanley R. Riddell ◽  
Hans-Peter Kiem
Keyword(s):  
Stem Cells ◽  
Gene Transfer ◽  
Cell Line ◽  
Flow Cytometric Analysis ◽  
Lentiviral Vectors ◽  
Transduction Efficiency ◽  
Packaging Cell Line ◽  
Hematopoietic Stem ◽  
Packaging Cell ◽  
Cd34 Cells

Vector-containing medium harvested from murine packaging cell lines has been shown to contain factors that can negatively influence the transduction and maintenance of hematopoietic stem cells. Thus, we generated a human packaging cell line with a gibbon ape leukemia virus pseudotype (Phoenix-GALV), and we evaluated vectors produced by Phoenix-GALV for their ability to transduce hematopoietic progenitor/stem cells. In 3 baboons, we used a competitive repopulation assay to directly compare GALV-pseudotype retrovirus vectors produced by either Phoenix-GALV or by the NIH 3T3–derived packaging cell line, PG13. In 3 additional baboons we compared Phoenix-GALV–derived vectors to more recently developed lentiviral vectors. Gene transfer efficiency into hematopoietic repopulating cells was assessed by evaluating the number of genetically modified peripheral blood and marrow cells using flow cytometry and real-time polymerase chain reaction. Transduction efficiency of hematopoietic repopulating cells was significantly higher using the Phoenix-GALV–derived vector as compared with the PG13-derived vectors or lentiviral vectors, with stable transduction levels up to 25%. We followed 2 animals for more than one year. Flow cytometric analysis of hematopoietic subpopulations in these animals revealed transgene expression in CD13+ granulocytes, CD20+ B lymphocytes, CD3+ T lymphocytes, CD61+ platelets, as well as red blood cells, indicating multilineage engraftment of cells transduced by Phoenix-GALV–pseudotype vectors. In addition, transduction of human CD34+ cells was significantly more efficient than transduction of baboon CD34+ cells, suggesting that Phoenix-GALV–derived oncoretroviral vectors may be even more efficient in human stem cell gene therapy applications.

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