Abstract
Pseudotyping lentivirus-based vectors is a strategy used to study conferred vector tropism and mechanisms of envelope glycoprotein function. Lentiviruses and filoviruses both assemble at the plasma membrane and have homotrimeric structural envelope glycoproteins that mediate both receptor binding and fusion. Such similarities help foster efficient pseudotyping. Importantly, filovirus glycoprotein pseudotyping of lentiviral vectors allows investigators to study virus entry at substantially less restrictive levels of biosafety containment than that required for wild-type filovirus work (biosafety level-2 vs. biosafety level-4, respectively). Standard lentiviral vector production involves transient transfection of viral component expression plasmids into producer cells, supernatant collection, and centrifuge concentration. Because the envelope glycoprotein expression plasmid is provided in trans, wild type or variant filoviral glycoproteins from marburgvirus or ebolavirus species may be used for pseudotyping and compared side-by-side. In this chapter we discuss the manufacture of pseudotyped lentiviral vector with an emphasis on small-scale laboratory grade production.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Naldini L, Blomer U, Gallay P, Ory D, Mulligan R, Gage FH, Verma IM, Trono D (1996) In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272:263–267
Sanders DA (2002) No false start for novel pseudotyped vectors. Curr Opin Biotechnol 13(5):437–442
Cronin J, Zhang XY, Reiser J (2005) Altering the tropism of lentiviral vectors through pseudotyping. Curr Gene Ther 5(4):387–398
Sena-Esteves M, Tebbets JC, Steffens S, Crombleholme T, Flake AW (2004) Optimized large-scale production of high titer lentivirus vector pseudotypes. J Virol Methods 122(2):131–139. doi:10.1016/j.jviromet.2004.08.017
Hislop JN, Islam TA, Eleftheriadou I, Carpentier DC, Trabalza A, Parkinson M, Schiavo G, Mazarakis ND (2014) Rabies virus envelope glycoprotein targets lentiviral vectors to the axonal retrograde pathway in motor neurons. J Biol Chem 289(23):16148–16163. doi:10.1074/jbc.M114.549980
Zhang C, Hu B, Xiao L, Liu Y, Wang P (2014) Pseudotyping lentiviral vectors with lymphocytic choriomeningitis virus glycoproteins for transduction of dendritic cells and in vivo immunization. Hum Gene Ther Methods 25(6):328–338. doi:10.1089/hgtb.2014.105
Beyer WR, Westphal M, Ostertag W, von Laer D (2002) Oncoretrovirus and lentivirus vectors pseudotyped with lymphocytic choriomeningitis virus glycoprotein: generation, concentration, and broad host range. J Virol 76(3):1488–1495
Sung VM, Lai MM (2002) Murine retroviral pseudotype virus containing hepatitis B virus large and small surface antigens confers specific tropism for primary human hepatocytes: a potential liver-specific targeting system. J Virol 76(2):912–917
Hsu M, Zhang J, Flint M, Logvinoff C, Cheng-Mayer C, Rice CM, McKeating JA (2003) Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles. Proc Natl Acad Sci U S A 100(12):7271–7276. doi:10.1073/pnas.0832180100
Funke S, Schneider IC, Glaser S, Muhlebach MD, Moritz T, Cattaneo R, Cichutek K, Buchholz CJ (2009) Pseudotyping lentiviral vectors with the wild-type measles virus glycoproteins improves titer and selectivity. Gene Ther 16(5):700–705. doi:10.1038/gt.2009.11
Cheng H, Koning K, O'Hearn A, Wang M, Rumschlag-Booms E, Varhegyi E, Rong L (2015) A parallel genome-wide RNAi screening strategy to identify host proteins important for entry of Marburg virus and H5N1 influenza virus. Virol J 12:194. doi:10.1186/s12985-015-0420-3
Kumar M, Bradow BP, Zimmerberg J (2003) Large-scale production of pseudotyped lentiviral vectors using baculovirus gp64. Hum Gene Ther 14:67–77
Sinn PL, Cooney AL, Oakland M, Dylla DE, Wallen TJ, Pezzulo AA, Chang EH, McCray PB Jr (2012) Lentiviral vector gene transfer to porcine airways. Mol Ther Nucleic Acids 1:e56. doi:10.1038/mtna.2012.47
Kang Y, Stein CS, Heth JA, Sinn PL, Penisten AK, Staber PD, Ratliff KL, Shen H, Barker CK, Martins I, Sharkey CM, Sanders DA, McCray PB Jr, Davidson BL (2002) In vivo gene transfer using a nonprimate lentiviral vector pseudotyped with ross river virus glycoproteins. J Virol 76:9378–9388
Kolokoltsov AA, Weaver SC, Davey RA (2005) Efficient functional pseudotyping of oncoretroviral and lentiviral vectors by Venezuelan equine encephalitis virus envelope proteins. J Virol 79(2):756–763. doi:10.1128/JVI.79.2.756-763.2005
Kobinger GP, Weiner DJ, Yu QC, Wilson JM (2001) Filovirus-pseudotyped lentiviral vector can efficiently and stably transduce airway epithelia in vivo. Nat Biotechnol 19(3):225–230. doi:10.1038/85664
Sinn PL, Hickey MA, Staber PD, Dylla DE, Jeffers SA, Davidson BL, Sanders DA, McCray PB Jr (2003) Lentivirus vectors pseudotyped with filoviral envelope glycoproteins transduce airway epithelia from the apical surface independently of folate receptor alpha. J Virol 77(10):5902–5910
Limberis MP, Bell CL, Heath J, Wilson JM (2010) Activation of transgene-specific T cells following lentivirus-mediated gene delivery to mouse lung. Mol Ther 18(1):143–150. doi:10.1038/mt.2009.190
Kondratowicz AS, Lennemann NJ, Sinn PL, Davey RA, Hunt CL, Moller-Tank S, Meyerholz DK, Rennert P, Mullins RF, Brindley M, Sandersfeld LM, Quinn K, Weller M, McCray PB Jr, Chiorini J, Maury W (2011) T-cell immunoglobulin and mucin domain 1 (TIM-1) is a receptor for Zaire Ebolavirus and Lake Victoria Marburgvirus. Proc Natl Acad Sci U S A 108(20):8426–8431. doi:10.1073/pnas.1019030108
Brindley MA, Hunt CL, Kondratowicz AS, Bowman J, Sinn PL, McCray PB Jr, Quinn K, Weller ML, Chiorini JA, Maury W (2011) Tyrosine kinase receptor Axl enhances entry of Zaire ebolavirus without direct interactions with the viral glycoprotein. Virology 415(2):83–94. doi:10.1016/j.virol.2011.04.002
Alvarez CP, Lasala F, Carrillo J, Muniz O, Corbi AL, Delgado R (2002) C-type lectins DC-SIGN and L-SIGN mediate cellular entry by Ebola virus in cis and in trans. J Virol 76(13):6841–6844
Watson DJ, Kobinger GP, Passini MA, Wilson JM, Wolfe JH (2002) Targeted transduction patterns in the mouse brain by lentivirus vectors pseudotyped with VSV, Ebola, Mokola, LCMV, or MuLV envelope proteins. Mol Ther 5(5 Pt 1):528–537. doi:10.1006/mthe.2002.0584
Medina MF, Kobinger GP, Rux J, Gasmi M, Looney DJ, Bates P, Wilson JM (2003) Lentiviral vectors pseudotyped with minimal filovirus envelopes increased gene transfer in murine lung. Mol Ther 8(5):777–789
Jeffers SA, Sanders DA, Sanchez A (2002) Covalent modifications of the ebola virus glycoprotein. J Virol 76(24):12463–12472
Sandrin V, Boson B, Salmon P, Gay W, Negre D, Le Grand R, Trono D, Cosset FL (2002) Lentiviral vectors pseudotyped with a modified RD114 envelope glycoprotein show increased stability in sera and augmented transduction of primary lymphocytes and CD34+ cells derived from human and nonhuman primates. Blood 100:823–832
Merten CA, Stitz J, Braun G, Poeschla EM, Cichutek K, Buchholz CJ (2005) Directed evolution of retrovirus envelope protein cytoplasmic tails guided by functional incorporation into lentivirus particles. J Virol 79(2):834–840. doi:10.1128/JVI.79.2.834-840.2005
Mammano F, Salvatori F, Indraccolo S, De Rossi A, Chieco-Bianchi L, Gottlinger HG (1997) Truncation of the human immunodeficiency virus type 1 envelope glycoprotein allows efficient pseudotyping of Moloney murine leukemia virus particles and gene transfer into CD4+ cells. J Virol 71(4):3341–3345
Trabalza A, Eleftheriadou I, Sgourou A, Liao TY, Patsali P, Lee H, Mazarakis ND (2014) Enhanced central nervous system transduction with lentiviral vectors pseudotyped with RVG/HIV-1gp41 chimeric envelope glycoproteins. J Virol 88(5):2877–2890. doi:10.1128/JVI.03376-13
Kang Y, Xia L, Tran DT, Stein CS, Hickey M, Davidson BL, McCray PB Jr (2005) Persistent expression of factor VIII in vivo following nonprimate letiviral gene transfer. Blood 106:1552–1558
Stremlau M, Owens CM, Perron MJ, Kiessling M, Autissier P, Sodroski J (2004) The cytoplasmic body component TRIM5alpha restricts HIV-1 infection in old world monkeys. Nature 427(6977):848–853. doi:10.1038/nature02343
Tuschong L, Soenen SL, Blaese RM, Candotti F, Muul LM (2002) Immune response to fetal calf serum by two adenosine deaminase-deficient patients after T cell gene therapy. Hum Gene Ther 13(13):1605–1610
Yamada K, McCarty DM, Madden VJ, Walsh CE (2003) Lentivirus vector purification using anion exchange HPLC leads to improved gene transfer. BioTechniques 34(5):1074–1078, 1080
Matsuoka H, Miyake K, Shimada T (1998) Improved methods of HIV vector mediated gene transfer. Int J Hematol 67(3):267–273
Slepushkin V, Chang N, Cohen R, Gan Y, Jiang B, Deausen E, Berlinger D, Binder G, Andre K, Humeau L, Dropulic B (2003) Large-scale purification of a lentiviral vector by size exclusion chromatography or mustang Q exchange capsule. Bioprocess J 2:89–95
Transfiguracion J, Jaalouk DE, Ghani K, Galipeau J, Kamen A (2003) Size-exclusion chromatography purification of high-titer vesicular stomatitis virus G glycoprotein-pseudotyped retrovectors for cell and gene therapy applications. Hum Gene Ther 14(12):1139–1153
Cooper AR, Patel S, Senadheera S, Plath K, Kohn DB, Hollis RP (2011) Highly efficient large-scale lentiviral vector concentration by tandem tangential flow filtration. J Virol Methods 177(1):1–9. doi:10.1016/j.jviromet.2011.06.019
Acknowledgments
This work was supported by the National Institutes of Health R01 HL105821 (PLS) and R21 AI123616 (WM). The University of Iowa Viral Vector Core is partially supported by the Cystic Fibrosis Foundation and the National Institutes of Health: P30 DK054759, P01 HL051670, and P30 CA086862.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Sinn, P.L., Coffin, J.E., Ayithan, N., Holt, K.H., Maury, W. (2017). Lentiviral Vectors Pseudotyped with Filoviral Glycoproteins. In: Hoenen, T., Groseth, A. (eds) Ebolaviruses. Methods in Molecular Biology, vol 1628. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7116-9_5
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7116-9_5
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7115-2
Online ISBN: 978-1-4939-7116-9
eBook Packages: Springer Protocols