Efforts to utilize genetically-modified T lymphocytes for adoptive T cell therapy have been limited by the relatively inefficient transduction efficiency obtained using retroviral vectors In preparation for utilizing the SIV/macaque model for in vivo studies of adoptive T cell therapy for AIDS, we have analyzed conditions affecting retroviral transduction of rhesus T cells To more accurately assess transduction efficiency, we utilized retroviral vectors encoding the murine CD2 molecule, thereby allowing precise determination of transduced cells by flow cytometry Transduction efficiency of rhesus lymphocytes using retroviral vector pseudotyped with the gibbon ape leukemia virus envelope was 5 to 10-fold greater as compared with vector containing the murine amphotropic envelope The introduction of centrifugation and phosphate depletion steps to our standard transduction protocol further enhanced transduction efficiency in NIH-3T3 or COS cells However, althoug h we can consistently enhance transduction efficiency upon phosphate starvation of various fibroblast cell lines, this step alone was not always effective for rhesus lymphocytes In fact, transduction efficiency of rhesus lymphocytes was significantly enhanced solely by centrifugation Results from these experiments will be used to design a protocol for the large scale transduction and expansion of genetically-modified rhesus T cells for use in in vivo studies in macaques

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Primate Research Center Grants (P51)
Project #
5P51RR000168-40
Application #
6453769
Study Section
Project Start
2001-05-01
Project End
2002-04-30
Budget Start
1997-10-01
Budget End
1998-09-30
Support Year
40
Fiscal Year
2001
Total Cost
$111,112
Indirect Cost
Name
Harvard University
Department
Type
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
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