Transduction efficiency of MLV but not of HIV-1 vectors is pseudotype dependent on human primary T lymphocytes

Abstract

The success of several gene therapeutic approaches requires efficient transduction of human primary T lymphocytes. For this it is important to enhance the transduction efficiency, and this can be achieved by various means, mainly technical development of transduction procedures and use of different vectors and vector pseudotypes. We analyzed the transduction efficiency of an HIV-1 vector encoding enhanced green fluorescent protein (GFP) as a marker gene and pseudotyped with the envelopes of MLV-A, MLV-10A1, GaLV, RD114, and VSV for human primary T lymphocytes in comparison to an MLV vector pseudotyped with the same envelopes. Pseudotyping of the MLV vector with the envelopes of 10A1 and GaLV resulted in efficient transduction of preactivated human primary T lymphocytes (32.4% and 32.7% CD3+/GFP+ cells, respectively) while MLV-A (14.0%), RD114 (8.8%), and VSV (1.5%) envelopes were less efficient when using titrated vector stocks equilibrated to a multiplicity of infection of 1. In contrast, the HIV-1 vectors pseudotyped with these envelope proteins transduced preactivated T lymphocytes with similar efficiency (approx. 20% CD3+/GFP+ cells). Thereby, CD4+ and CD8+ T lymphocyte subpopulations were transduced at equivalent levels. The similar performance of the different HIV-1 vector pseudotypes may be due in part to the similar half-lives of the vector particles. Independently of the envelope used for pseudotyping neither the MLV nor the HIV-1 vectors yielded any significant transduction in nonactivated T lymphocytes (below 0.55% of GFP+ cells)

Transduction of Nondividing Human Macrophages with Gammaretrovirus-Derived Vectors

“…Lymphocytes were treated with either PHA plus IL-2 or with IL-7, which induce cell proliferation or G 0 /G 1b transition but induced only marginal cell proliferation under the conditions used here, respectively (4,11). Dividing PHA-IL-2-stimulated lymphocytes were susceptible to transduction with both retroviral vectors, although the percentage of F-MLV-transduced cells was lower than that of HIV-1-transduced cells, as reported by others (26). On the contrary, although IL-7-treated lymphocytes were susceptible to HIV-1 transduction, albeit with variations depending on the donor, they were largely resistant to F-MLV and were transduced at rates below 1% for a multiplicity of infection (MOI) of 25.…”

Altering the Tropism of Lentiviral Vectors through Pseudotyping

“…In addition, amphotropic MLV pseudotypes transduced cytokine-mobilized, human peripheral blood CD34+ cells capable of establishing hematopoiesis in immunodeficient mice more efficiently than VSV-G pseudotypes. Work reported by Muhlebach et al [Muhlebach, 2003] revealed that MLV vectors pseudotyped with the MLV 10A1 and the GALV GPs resulted in efficient transduction of preactivated human primary T lymphocytes while pseudotypes bearing the amphotropic MLV, RD114 and VSV-G GPs were less efficient. In contrast, HIV-1 vectors pseudotyped with the same GPs transduced preactivated T lymphocytes with similar efficiencies.…”

Redirecting T lymphocyte specificity by T cell receptor gene transfer – A new era for immunotherapy

“…The high transduction efficiency of primary human T cells using the MLV-10A1 pseudotype is based on the usage of both Pit1 and Pit2 for cell entry Miller and Chen, 1996;Uckert et al, 1998). There are contrasting reports with regard to the transduction efficiency of human T cells achieved with the endogenous feline retrovirus RD114 pseudotype (Porter et al, 1996;Onodera et al, 1998;Uckert et al, 2000;Muhlebach et al, 2003;Strom et al, 2003). High transduction efficiencies obtained with the envelope of vesicular stomatitis virus G protein (VSV-G) are most probably due to the use of high multiplicities of infection (MOI).…”

“…High transduction efficiencies obtained with the envelope of vesicular stomatitis virus G protein (VSV-G) are most probably due to the use of high multiplicities of infection (MOI). When VSV-G pseudotyped retrovirus vectors were carefully equilibrated in titer to other pseudotypes transduction efficiency dramatically decreased (Muhlebach et al, 2003). Primary murine T cells are most efficiently transduced by retroviral vectors carrying the ecotropic MLV envelope (Hagani et al, 1999;Engels et al, 2003).…”