Efficiency of onco-retroviral and lentiviral gene transfer into primary mouse and human B-lymphocytes is pseudotype dependent.

B lymphocytes are attractive targets for gene therapy of genetic diseases associated with B-cell dysfunction and for immunotherapy. Transduction of B lymphocytes was evaluated using green fluorescent protein (GFP)-encoding onco-retroviral and HIV-derived lentiviral vectors which were pseudotyped with ecotropic, amphotropic or vesicular stomatitis virus (VSV-G) envelopes. Transduction of mouse B lymphocytes activated with lipopolysaccharides (LPS) or by cross-linking CD40 in conjunction with interleukin-4 (IL-4) was significantly more efficient (p < 0.003) with ecotropic (11%) than with VSV-G pseudotyped onco-retroviral vectors (1%). Using high-titer cell-free ecotropic viral supernatant or by coculture with ecotropic onco-retroviral vector-producing cells, transduction efficiency increased significantly (p < 0.001) to approximately 50%, whereas transduction efficiency by coculture with VSV-G pseudotyped vector-producing cells remained low (< 2%). Similarly, transduction of mouse B lymphocytes was significantly more efficient (twofold, p < 0.01) with the ecotropic (7%) than with the VSV-G pseudotyped lentiviral vectors although gene transfer efficiency remained low because of dose-limiting toxicity of the concentrated vector preparations on the LPS-activated murine B cells. Consistent with murine B-cell transduction, human B cells activated with CD40L and IL-4 were also found to be relatively refractory to VSV-G pseudotyped onco-retroviral vectors (< 1%). However, higher transduction efficiencies could be achieved in activated primary human B lymphocytes using VSV-G pseudotyped lentiviral vectors instead (5%-6%). Contrary to the significant increase in mouse B-cell transduction efficiency with ecotropic vectors, the use of amphotropic onco-retroviral or lentiviral vectors did not increase transduction efficiency in primary human B cells. The present study shows that the transduction efficiency of onco-retroviral and lentiviral vectors in human and mouse B lymphocytes is pseudotype-dependent and challenges the widely held assumption that VSV-G pseudotyping facilitates gene transfer into all cell types.

[1]  L. Naldini,et al.  Molecular evidence of inefficient transduction of proliferating human B lymphocytes by VSV-pseudotyped HIV-1-derived lentivectors. , 2004, Virology.

[2]  G. Kobinger,et al.  Transduction of human islets with pseudotyped lentiviral vectors. , 2004, Human gene therapy.

[3]  A. Sheriff,et al.  Efficient in vitro transduction of naive murine B cells with lentiviral vectors. , 2004, Biochemical and biophysical research communications.

[4]  L. Naldini,et al.  Lentiviral vectors containing the human immunodeficiency virus type-1 central polypurine tract can efficiently transduce nondividing hepatocytes and antigen-presenting cells in vivo. , 2002, Blood.

[5]  G. Kobinger,et al.  Targeted transduction patterns in the mouse brain by lentivirus vectors pseudotyped with VSV, Ebola, Mokola, LCMV, or MuLV envelope proteins. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[6]  S. S. Case,et al.  Lentiviral vector gene transfer into fetal rhesus monkeys (Macaca mulatta): lung-targeting approaches. , 2001, Molecular therapy : the journal of the American Society of Gene Therapy.

[7]  A. Keegan,et al.  Efficient transduction of murine B lymphocytes and B lymphoma lines by modified adenoviral vectors: enhancement via targeting to FcR and heparan-containing proteins , 2001, Gene Therapy.

[8]  D. Rowe,et al.  Use of VSV-G pseudotyped retroviral vectors to target murine osteoprogenitor cells. , 2001, Virology.

[9]  Y. Ron,et al.  An inducible packaging cell system for safe, efficient lentiviral vector production in the absence of HIV-1 accessory proteins. , 2001, Virology.

[10]  Y. Ron,et al.  A gene therapy approach for treating T-cell-mediated autoimmune diseases. , 2001, Blood.

[11]  D. Scott,et al.  Mechanisms of Tolerance Induction by a Gene-Transferred Peptide-IgG Fusion Protein Expressed in B Lineage Cells1 2 , 2000, The Journal of Immunology.

[12]  D. Bodine,et al.  Lentivirus-based vectors transduce mouse hematopoietic stem cells with similar efficiency to moloney murine leukemia virus-based vectors. , 2000, Blood.

[13]  S. Chang,et al.  Generation of retroviral packaging and producer cell lines for large-scale vector production and clinical application: improved safety and high titer. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[14]  R. Morgan,et al.  Lentiviral-mediated gene transfer into human lymphocytes: role of HIV-1 accessory proteins. , 2000, Blood.

[15]  W. Uckert,et al.  Lentiviral vectors pseudotyped with envelope glycoproteins derived from gibbon ape leukemia virus and murine leukemia virus 10A1. , 2000, Virology.

[16]  J. Dick,et al.  Transduction of human CD34+ CD38- bone marrow and cord blood-derived SCID-repopulating cells with third-generation lentiviral vectors. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[17]  L. Ailles,et al.  Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences , 2000, Nature Genetics.

[18]  Y. Takeuchi,et al.  Progress with retroviral gene vectors , 2000, Reviews in medical virology.

[19]  A. Miller,et al.  Superior transduction of mouse hematopoietic stem cells with 10A1 and VSV-G pseudotyped retrovirus vectors. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[20]  B. Barnhart,et al.  A Transcriptional Defect Underlies B Lymphocyte Dysfunction in a Patient Diagnosed with Non-X-Linked Hyper-IgM Syndrome1 , 2000, The Journal of Immunology.

[21]  D. Scott,et al.  Retroviral gene therapy with an immunoglobulin-antigen fusion construct protects from experimental autoimmune uveitis. , 2000, The Journal of clinical investigation.

[22]  B. Rapoport,et al.  Autoimmune Response to the Thyroid in Humans: Thyroid Peroxidase-The Common Autoantigenic Denominator , 2000, International reviews of immunology.

[23]  W. Anderson,et al.  High efficiency in vitro gene transfer into vascular tissues using a pseudotyped retroviral vector without pseudotransduction , 1999, Gene Therapy.

[24]  M. Sadelain,et al.  Activation conditions determine susceptibility of murine primary T‐lymphocytes to retroviral infection , 1999, The journal of gene medicine.

[25]  T. VandenDriessche,et al.  Long-term expression of human coagulation factor VIII and correction of hemophilia A after in vivo retroviral gene transfer in factor VIII-deficient mice. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[26]  D. Jolly,et al.  Animal Testing of Retroviral-Mediated Gene Therapy for Factor VIII Deficiency , 1999, Thrombosis and Haemostasis.

[27]  H. Iba,et al.  Dose-dependent transduction of vesicular stomatitis virus G protein-pseudotyped retrovirus vector into human solid tumor cell lines and murine fibroblasts. , 1999, Virology.

[28]  W. Anderson,et al.  Receptor-Mediated Moloney Murine Leukemia Virus Entry Can Occur Independently of the Clathrin-Coated-Pit-Mediated Endocytic Pathway , 1999, Journal of Virology.

[29]  D. Trono,et al.  Self-Inactivating Lentivirus Vector for Safe and Efficient In Vivo Gene Delivery , 1998, Journal of Virology.

[30]  K. Weinberg,et al.  Gene delivery to human B-precursor acute lymphoblastic leukemia cells. , 1998, Blood.

[31]  M F Hoylaerts,et al.  Mechanism and kinetics of factor VIII inactivation: study with an IgG4 monoclonal antibody derived from a hemophilia A patient with inhibitor. , 1998, Blood.

[32]  H. Brems,et al.  Bone marrow stromal cells as targets for gene therapy of hemophilia A. , 1998, Human gene therapy.

[33]  M. Sadelain,et al.  Recombinant retroviruses pseudotyped with the vesicular stomatitis virus G glycoprotein mediate both stable gene transfer and pseudotransduction in human peripheral blood lymphocytes. , 1997, Blood.

[34]  R. Morgan,et al.  Efficient in vivo marking of primary CD4+ T lymphocytes in nonhuman primates using a gibbon ape leukemia virus-derived retroviral vector. , 1997, Blood.

[35]  Elias T. Zambidis,et al.  Genetically Transferred Central and Peripheral Immune Tolerance via Retroviral-Mediated Expression of Immunogenic Epitopes in Hematopoietic Progenitors or Peripheral B Lymphocytes , 1997, Molecular medicine.

[36]  A. Miller.  Cell-surface receptors for retroviruses and implications for gene transfer. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[37]  R. Mulligan,et al.  A stable human-derived packaging cell line for production of high titer retrovirus/vesicular stomatitis virus G pseudotypes. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[38]  F. Gage,et al.  In Vivo Gene Delivery and Stable Transduction of Nondividing Cells by a Lentiviral Vector , 1996, Science.

[39]  M. Kay,et al.  Pseudotransduction of hepatocytes by using concentrated pseudotyped vesicular stomatitis virus G glycoprotein (VSV-G)-Moloney murine leukemia virus-derived retrovirus vectors: comparison of VSV-G and amphotropic vectors for hepatic gene transfer , 1996, Journal of virology.

[40]  K. Weinberg,et al.  Multiple modifications in cis elements of the long terminal repeat of retroviral vectors lead to increased expression and decreased DNA methylation in embryonic carcinoma cells , 1995, Journal of virology.

[41]  O. Danos,et al.  Receptor-binding domain of murine leukemia virus envelope glycoproteins , 1995, Journal of virology.

[42]  J. Burns,et al.  A general method for the generation of high-titer, pantropic retroviral vectors: highly efficient infection of primary hepatocytes. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[43]  Y. Ron,et al.  A murine model for B-lymphocyte somatic cell gene therapy. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[44]  Wei Wei Wu,et al.  Monoclonal antibodies to murine CD40 define two distinct functional epitopes , 1994, European journal of immunology.

[45]  W. Anderson,et al.  pH-independent murine leukemia virus ecotropic envelope-mediated cell fusion: implications for the role of the R peptide and p12E TM in viral entry , 1994, Journal of virology.

[46]  D. Campana,et al.  X‐Linked Agammaglobulinemia: New Approaches to Old Questions based on the Identification of the Defective Gene , 1994, Immunological reviews.

[47]  K. Nagashima,et al.  Function of the cytoplasmic domain of a retroviral transmembrane protein: p15E-p2E cleavage activates the membrane fusion capability of the murine leukemia virus Env protein , 1994, Journal of virology.

[48]  R. Edwards,et al.  Cloning of the cellular receptor for amphotropic murine retroviruses reveals homology to that for gibbon ape leukemia virus. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[49]  K. Gilbert,et al.  Tolerogenicity of resting and activated B cells , 1994, The Journal of experimental medicine.

[50]  W. Anderson,et al.  Sequences determining the pH dependence of viral entry are distinct from the host range-determining region of the murine ecotropic and amphotropic retrovirus envelope proteins , 1993, Journal of virology.

[51]  D. Parker,et al.  Parameters of tolerance induction by antigen targeted to B lymphocytes. , 1993, Journal of immunology.

[52]  J. Banchereau,et al.  Growing human B lymphocytes in the CD40 system , 1991, Nature.

[53]  R. North,et al.  Cell-surface receptor for ecotropic murine retroviruses is a basic amino-acid transporter , 1991, Nature.

[54]  S. Ziegler,et al.  Efficient gene transfer and expression in primary B lymphocytes. , 1991, Journal of immunological methods.

[55]  R. Weiss,et al.  The pH independence of mammalian retrovirus infection. , 1990, The Journal of general virology.

[56]  S. Goff,et al.  A safe packaging line for gene transfer: separating viral genes on two different plasmids , 1988, Journal of virology.

[57]  J. Dick,et al.  High efficiency gene transfer and expression in normal murine B lymphocytes. , 1987, Journal of immunological methods.

[58]  A. Miller,et al.  Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production , 1986, Molecular and cellular biology.

[59]  B. Nexø,et al.  Entry of murine retrovirus into mouse fibroblasts. , 1983, Virology.

[60]  I. Pastan,et al.  Inhibition of VSV binding and infectivity by phosphatidylserine: Is phosphatidylserine a VSV-binding site? , 1983, Cell.

[61]  L. Naldini,et al.  Oncoretroviral and lentiviral vector-mediated gene therapy. , 2002, Methods in enzymology.

[62]  I. Verma,et al.  Generation of a stable cell line producing high-titer self-inactivating lentiviral vectors. , 2001, Molecular therapy : the journal of the American Society of Gene Therapy.

[63]  M. Kay,et al.  Efficient lentiviral transduction of liver requires cell cycling in vivo , 2000, Nature Genetics.

[64]  M. Yamada,et al.  Successful peripheral T-lymphocyte-directed gene transfer for a patient with severe combined immune deficiency caused by adenosine deaminase deficiency. , 1998, Blood.