1. Volume 18, Issue 4, Pages 725-733 (April 2010)
Cocal-pseudotyped Lentiviral Vectors Resist Inactivation by Human Serum and Efficiently Transduce Primate Hematopoietic Repopulating Cells 
Grant D Trobridge, Robert A Wu, Michael Hansen, Christina Ironside, Korashon L Watts, Philip Olsen, Brian C Beard, Hans-Peter Kiem 
Molecular Therapy 
Volume 18, Issue 4, Pages (April 2010)
DOI: /mt
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

2. Figure 1
Envelope constructs and cell tropism. (a) Envelope plasmids. All envelopes were expressed from a cytomegalovirus (CMV) promoter with a human or rabbit β-globin intron (hBGint, rBGint) 5′ to the open reading frame (ORF) and a human or rabbit β-globin poly A sequence (hBGpA, rBGpA). The cocal ORF was codon-optimized for human cells. (b) Tropism of cocal envelope relative to vesicular stomatitis virus envelope glycoprotein (VSV-G) and RD114/TR envelopes. The indicated cell lines and primary cell cultures were transduced at a multiplicity of infection of 5 and the percentage of enhanced green fluorescent protein (EGFP)-expressing cells was determined 6 days after vector exposure.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

3. Figure 2
Serum neutralization of lentiviral pseudotypes. Serum from (a) 10 individuals, (b) 5 dogs, and (c) 5 macaques was incubated with vector at 37 °C for 30 minutes then added to HT1080 cells to evaluate the number of enhanced green fluorescent protein (EGFP) transducing units. EGFP expression was evaluated by flow cytometry 3 days after vector exposure, and the percentage of EGFP-expressing cells after incubation in the serum was determined relative to the percentage of EGFP-expressing cells in the vector-only control to determine the fold increase or decrease in titer after exposure to serum. VSV-G, vesicular stomatitis virus envelope glycoprotein.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

4. Figure 3
Comparison of pseudotypes for gene transfer efficiency to hematopoietic progenitors. (a) Human CD34+ cells were exposed to vector for 20 hours and then added to each well of a 24-well plate for colony forming unit (CFU) analysis. The percentage of enhanced green fluorescent protein (EGFP)-expressing CFUs was determined by fluorescent microscopy 14 days after vector exposure. (b) Pigtailed macaque CD34+ cells from three donors were exposed to vector at a multiplicity of infection (MOI) of 5 and the percentage of EGFP-expressing cells was determined 10 days after vector exposure. (c) Dog CD34+ cells from three donors were exposed to vector at an MOI of 5 and the percentage of EGFP-expressing cells was determined 10 days after vector exposure. The mean and the SE from three replicates are shown. VSV-G, vesicular stomatitis virus envelope glycoprotein.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

5. Figure 4
Comparison of cocal and vesicular stomatitis virus envelope glycoprotein (VSV-G)-mediated gene transfer to human CD34+ severe combined immunodeficient–repopulating cells (SRCs). The mean marking for the cocal-pseudotyped enhanced green fluorescent protein (EGFP)-expressing vector and the VSV-G-pseudotyped EYFP-expressing vector is shown in pretransplant liquid cultures, progenitor cultures, and in human SRCs from mouse bone marrow (BM) 5 weeks after transplantation as determined by flow cytometry. The mean engraftment of human CD45+ cells as determined by flow cytometry is also shown in mouse BM at 5 weeks. The SE from five transplanted mice is shown. CFU, colony forming unit.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

6. Figure 5
Transgene expression from cocal and vesicular stomatitis virus envelope glycoprotein (VSV-G)-pseudotyped vectors in macaque peripheral blood cells. (a) The percentages of enhanced green fluorescent protein (EGFP) (cocal) and enhanced yellow fluorescent protein (EYFP) (VSV-G)-expressing leukocytes in peripheral blood detected by flow cytometry are shown. (b) A representative example of the gating used to distinguish EGFP- and EYFP-expressing leukocytes.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions

7. Figure 6
Comparison of cocal and vesicular stomatitis virus envelope glycoprotein (VSV-G)-mediated gene transfer in peripheral blood leukocyte subpopulations and bone marrow (BM) CD34+ cells. The percentage of leukocytes that (a) express enhanced green fluorescent protein (EGFP) (cocal) or (b) express enhanced yellow fluorescent protein (EYFP) (VSV-G) in different leukocyte subpopulations was determined using phycoerythrin (PE)-labeled lineage-specific antibodies. PB CD3+ T lymphocytes, CD20+ B lymphocytes, and CD14+ myeloid cells are shown at day 104 after transplantation. For each subset either EYFP- (a), or EGFP- (b) positive cells in this subset are excluded by gating and the percentage of cocal EGFP-positive cells (a) or VSV-G EYFP-positive cells (b) are shown. (c) The percentage of EYFP/EGFP cells in each lineage is graphed for each experimental arm. For both pseudotypes, transgene-expressing cells were found in all lineages examined at similar frequencies relative to BM CD34+ cells.
Molecular Therapy  , DOI: ( /mt )
Copyright © 2010 The American Society of Gene & Cell Therapy Terms and Conditions