Ex vivo expansion and lentiviral transduction of Macaca nemestrina CD4+ T cells.

ABSTRACT

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.