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Recombinant fowlpox virus for in vitro gene delivery to pancreatic islet tissue
Author(s) -
Solomon Michelle F,
Ramshaw Ian A,
Simeonovic Charmaine J
Publication year - 2005
Publication title -
immunology and cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0818-9641
DOI - 10.1111/j.1440-1711.2005.01379.x
Subject(s) - fowlpox virus , recombinant dna , islet , in vitro , virology , biology , gene , virus , gene delivery , microbiology and biotechnology , genetic enhancement , genetics , endocrinology , insulin
The feasibility of using avipox virus as a vector for gene delivery to islet tissue (adult islets and fetal proislets) was examined using a recombinant fowlpox virus (FPV) engineered to express the reporter gene LacZ (FPV‐LacZ). The efficiency of in vitro transduction was dose‐dependent and influenced by the donor species and maturation status of the islet tissue. Reporter gene expression in FPV‐LacZ‐transduced islet grafts was transient (3ndash;7 days) in immunoincompetent nude mice and was not prolonged by in vivo treatment with anti‐IFN‐γ mAb. In contrast, FPV‐LacZ‐transduced NIT‐1 cells (a mouse islet beta cell line) expressed the LacZ gene beyond 18 days in vitro . Silencing of transgene expression therefore appeared to occur in vivo and was T cell‐ and IFN‐γ‐independent. Isografts of FPV‐LacZ‐transduced islets in immunocompetent mice underwent immunological destruction by 7 days, suggesting that either FPV proteins or the reporter protein β‐galactosidase induced an adaptive immune response. Co‐delivery of the rat bioactive immunoregulatory cytokine gene TGF‐β to islets using FPV‐TGF‐β led to enhanced expression of TGF‐β mRNA in isografts but no long‐term protection. Nevertheless, compared to control islet isografts at 5 days, FPV‐transduced islets remained embedded in the clotted blood used to facilitate implantation. This phenomenon was TGF‐β transgene‐independent, correlated with lack of cellular infiltration, and suggested that the FPV vector transformed the blood clot into a temporary immunological barrier.