Infection of intact human islets by a lentiviral vector.
The transfer of genes encoding immunomodulatory proteins to islets can be used to improve islet function, block apoptosis, and inhibit rejection following transplantation. Adenoviral vectors have been shown to infect intact human islets, but the immunogenicity and transient gene expression of the current adenoviral vectors may hinder their use clinically for islet transplantation. In this report, we compared an HIV-1-based lentiviral vector with the E1-deleted adenoviral vehicle of the Ad5 type for gene transfer to human islets in vitro. We demonstrate that at similar viral particle concentrations per islet that an HIV-based lentiviral vector is able to infect beta-cells within an intact human islet at an efficiency similar to an adenoviral vector. In addition, both the adenoviral and lentiviral vectors were able to express significant levels of soluble interleukin-1 receptor antagonist (IL-1Ra) protein following infection of intact islets. More importantly, there was no impairment of islet beta-cell function following adenoviral and lentiviral infection in responding to glucose stimulation. These results support the utility of replication-defective lentiviral vectors as efficient gene delivery vehicles to islets to faciliate transplantation of islets for therapy of type I diabetes.
Giannoukakis
N
N
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Mi
Z
Z
Gambotto
A
A
Eramo
A
A
Ricordi
C
C
Trucco
M
M
Robbins
P
P
D000256:Adenoviridae / Q000235:genetics; D017209:Apoptosis; D002478:Cells, Cultured; D003673:Defective Viruses; D005434:Flow Cytometry; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005947:Glucose / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D053590:Interleukin 1 Receptor Antagonist Protein; D007515:Islets of Langerhans / Q000378:metabolism / Q000821:virology*; D008164:Luminescent Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D012795:Sialoglycoproteins / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
C507763:IL1RN protein, human; D007328:Insulin; D053590:Interleukin 1 Receptor Antagonist Protein; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D012795:Sialoglycoproteins; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase; D005947:Glucose