Ex vivo expansion and lentiviral transduction of Macaca nemestrina CD4+ T cells.
BACKGROUND
Macaca nemestrina is a nonhuman primate used as a model in preclinical studies of hematopoietic stem cell transplantation and adoptive transfer of T cells. Adoptive T cell transfer studies typically require ex vivo expansion of substantial numbers of T cells prior to their reinfusion into the subject.


METHODS
Pigtailed macaque peripheral blood CD4(+) cells were expanded using CD3 and CD28 antibody-coated beads. These cells were transformed using Herpesvirus saimiri and were also transduced with HIV-1 based lentiviral vectors.


RESULTS
We report an efficient method for the ex vivo expansion of CD4(+) T cells from Macaca nemestrina peripheral blood. With this protocol, primary CD4(+) T cells can be expanded between 300- to 6000-fold during 24-day period and can be efficiently transduced with lentiviral vectors. Furthermore, these T cells can be transformed by Herpesvirus saimiri and maintained in culture for several months. The transformed T cell lines can be productively infected with the simian immunodeficiency virus (SIV) strain SIV(mac239).


CONCLUSIONS
We have established methods for the expansion and transformation of primary M. nemestrina CD4(+) T cells and demonstrated the utility of these methods for several applications.
Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
Nina M
NM
Muñoz
Grant D
GD
Trobridge
Hans-Peter
HP
Kiem
D000818:Animals; D018106:CD28 Antigens / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000821:virology; D018929:Cell Culture Techniques; D002461:Cell Line, Transformed; D006801:Humans; D018189:Immunomagnetic Separation; D008254:Macaca nemestrina / Q000276:immunology*; D016097:Simian Acquired Immunodeficiency Syndrome; D015302:Simian Immunodeficiency Virus
D018106:CD28 Antigens; D017252:CD3 Complex