Genetic modification of human peripheral blood lymphocytes with a transdominant negative form of Rev: safety and toxicity.
A transdominant mutant form of the rev gene, M10, confers resistance to infection by the human immunodeficiency virus (HIV) in vitro and is currently under investigation as a potential intervention in acquired immunodeficiency syndrome (AIDS). In this report, we examine three issues relevant to the safety of autologous transfer of human T cells genetically modified with Rev M10. First, the potential for malignant transformation was assessed in vitro using interleukin-2 (IL-2) dependence and fibroblast transformation assays, and tumorigenicity was evaluated in severe combined immunodeficient (SCID) mice. Possible toxicity was evaluated by pathologic analysis following adoptive transfer of genetically modified human T cells into SCID mice. Second, methods were developed that permit T cell activation required for gene transfer but do not allow replication of endogenous HIV. Third, T cell function was evaluated in peripheral blood lymphocytes (PBL) of HIV-seropositive donors transduced with Rev M10 and compared to a negative control mutant, delta Rev M10. By all criteria, no oncogenicity or toxicity was observed. Human T cells transduced with these vectors did not grow in the absence of IL-2 in vitro, and no tumors were observed following transplantation of genetically modified human cells into recipient SCID mice. Histopathological analysis of heart, lung, liver, spleen, and kidney of animals 1-21 weeks following adoptive transfer of gene-modified human T cells revealed no significant abnormalities. Additionally, no differences were observed in the pattern of cytokine secretion in enriched human PBL expressing Rev M10 compared to delta Rev M10. (ABSTRACT TRUNCATED AT 250 WORDS)
Fox
B A
BA
Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor 48109-0650, USA.
Woffendin
C
C
Yang
Z Y
ZY
San
H
H
Ranga
U
U
Gordon
D
D
Osterholzer
J
J
Nabel
G J
GJ
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D001483:Base Sequence; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D016207:Cytokines / Q000096:biosynthesis; D005260:Female; D005347:Fibroblasts; D018014:Gene Transfer Techniques; D005799:Genes, Dominant; D015757:Genes, rev / Q000235:genetics*; D005822:Genetic Vectors; D006046:Gold / Q000633:toxicity; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D006801:Humans; D016219:Immunotherapy, Adoptive; D007376:Interleukin-2 / Q000502:physiology; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D016513:Mice, SCID; D008863:Microspheres; D008969:Molecular Sequence Data; D012190:Retroviridae / Q000235:genetics; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism / Q000637:transplantation; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; D016207:Cytokines; D007376:Interleukin-2; D018894:Reverse Transcriptase Inhibitors; D006046:Gold