Inducible expression of the deltaNGFr/F12Nef fusion protein as a new tool for anti-human immunodeficiency virus type 1 gene therapy.
Expression of the human immunodeficiency virus type 1 (HIV-1) Nef triple mutant F12Nef strongly inhibits HIV-1 replication. We exploited such a unique feature in a novel anti-HIV-1 gene therapy design by constructing an HIV-1 Tat-defective lentivirus vector expressing the product of fusion between the low-affinity human nerve growth factor receptor truncated in its intracytoplasmic domain (deltaNGFr, NH(2) moiety), and F12Nef (COOH moiety), under the control of the HIV-1 long terminal repeats. In this manner, both the selection marker (deltaNGFr) and the anti-HIV-1 effector are comprised in the same fusion protein, the expression of which is targetable by HIV-1 infection. Such a vector was proved to transduce human cells efficiently and, on HIV-1 infection, it expressed high levels of the fusion protein. In addition, strong antiviral activity of the deltaNGFr/F12Nef-expressing vector was demonstrated in cell lines as well as in primary cell cultures challenged with T- or M-tropic HIV-1 isolates. Thus, the HIV-1-targetable expression of the deltaNGFr/F12Nef fusion protein represents a novel and powerful tool for an effective anti-HIV-1 gene therapy strategy.
Muratori
Claudia
C
Laboratory of Virology, Istituto Superiore di Sanità, 00161 Rome, Italy.
Schiavoni
Ilaria
I
Melucci-Vigo
Gianna
G
Olivetta
Eleonora
E
Santarcangelo
Anna Claudia
AC
Pugliese
Katherina
K
Verani
Paola
P
Federico
Maurizio
M
D015496:CD4-Positive T-Lymphocytes / Q000821:virology; D002460:Cell Line / Q000821:virology; D003673:Defective Viruses / Q000235:genetics / Q000502:physiology*; D015967:Gene Expression Regulation, Viral; D015754:Genes, nef; D015756:Genes, tat; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics* / Q000502:physiology; D015658:HIV Infections / Q000628:therapy*; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells / Q000821:virology; D006801:Humans; D007668:Kidney; D016086:Lentivirus / Q000235:genetics* / Q000502:physiology; D007963:Leukocytes, Mononuclear / Q000821:virology; D008213:Lymphocyte Activation; D008264:Macrophages / Q000821:virology; D009000:Monocytes / Q000166:cytology; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D014158:Transcription, Genetic; D014161:Transduction, Genetic; D014779:Virus Replication / Q000187:drug effects*
D011993:Recombinant Fusion Proteins; C472328:deltaNGFr-F12Nef fusion protein, recombinant