Cellular immune response against firefly luciferase after sleeping beauty-mediated gene transfer in vivo

Kelly M. Podetz-Pedersen, Vaiva Vezys, Nikunj V Somia, Stephen J. Russell, R S Mc Ivor

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The Sleeping Beauty (SB) transposon system has been shown to mediate new gene sequence integration resulting in long-term expression. Here the effectiveness of hyperactive SB100X transposase was tested, and we found that hydrodynamic co-delivery of a firefly luciferase transposon (pT2/CaL) along with SB100X transposase (pCMV-SB100X) resulted in remarkably sustained, high levels of luciferase expression. However, after 4 weeks there was a rapid, animal-by-animal loss of luciferase expression that was not observed in immunodeficient mice. We hypothesized that this sustained, high-level luciferase expression achieved using the SB100X transposase elicits an immune response in pT2/CaL co-administered mice, which was supported by the rapid loss of luciferase expression upon challenge of previously treated animals and in naive animals adoptively transferred with splenocytes from previously treated animals. Specificity of the immune response to luciferase was demonstrated by increased cytokine expression in splenocytes after exposure to luciferase peptide in parallel with MHC I-luciferase peptide tetramer binding. This anti-luciferase immune response observed following continuous, high-level luciferase expression in vivo clearly impacts its use as an in vivo reporter. As both an immunogen and an extremely sensitive reporter, luciferase is also a useful model system for the study of immune responses following in vivo gene transfer and expression.

Original languageEnglish (US)
Pages (from-to)955-965
Number of pages11
JournalHuman gene therapy
Volume25
Issue number11
DOIs
StatePublished - Nov 1 2014

Fingerprint

Firefly Luciferases
Beauty
Luciferases
Cellular Immunity
Genes
Transposases
Peptides
Hydrodynamics
Immune System
Cytokines

Cite this

Cellular immune response against firefly luciferase after sleeping beauty-mediated gene transfer in vivo. / Podetz-Pedersen, Kelly M.; Vezys, Vaiva; Somia, Nikunj V; Russell, Stephen J.; Mc Ivor, R S.

In: Human gene therapy, Vol. 25, No. 11, 01.11.2014, p. 955-965.

Research output: Contribution to journalArticle

@article{230cfcf448854647a8cd6acdaad3daf9,
title = "Cellular immune response against firefly luciferase after sleeping beauty-mediated gene transfer in vivo",
abstract = "The Sleeping Beauty (SB) transposon system has been shown to mediate new gene sequence integration resulting in long-term expression. Here the effectiveness of hyperactive SB100X transposase was tested, and we found that hydrodynamic co-delivery of a firefly luciferase transposon (pT2/CaL) along with SB100X transposase (pCMV-SB100X) resulted in remarkably sustained, high levels of luciferase expression. However, after 4 weeks there was a rapid, animal-by-animal loss of luciferase expression that was not observed in immunodeficient mice. We hypothesized that this sustained, high-level luciferase expression achieved using the SB100X transposase elicits an immune response in pT2/CaL co-administered mice, which was supported by the rapid loss of luciferase expression upon challenge of previously treated animals and in naive animals adoptively transferred with splenocytes from previously treated animals. Specificity of the immune response to luciferase was demonstrated by increased cytokine expression in splenocytes after exposure to luciferase peptide in parallel with MHC I-luciferase peptide tetramer binding. This anti-luciferase immune response observed following continuous, high-level luciferase expression in vivo clearly impacts its use as an in vivo reporter. As both an immunogen and an extremely sensitive reporter, luciferase is also a useful model system for the study of immune responses following in vivo gene transfer and expression.",
author = "Podetz-Pedersen, {Kelly M.} and Vaiva Vezys and Somia, {Nikunj V} and Russell, {Stephen J.} and {Mc Ivor}, {R S}",
year = "2014",
month = "11",
day = "1",
doi = "10.1089/hum.2014.048",
language = "English (US)",
volume = "25",
pages = "955--965",
journal = "Human Gene Therapy",
issn = "1043-0342",
publisher = "Mary Ann Liebert Inc.",
number = "11",

}

TY - JOUR

T1 - Cellular immune response against firefly luciferase after sleeping beauty-mediated gene transfer in vivo

AU - Podetz-Pedersen, Kelly M.

AU - Vezys, Vaiva

AU - Somia, Nikunj V

AU - Russell, Stephen J.

AU - Mc Ivor, R S

PY - 2014/11/1

Y1 - 2014/11/1

N2 - The Sleeping Beauty (SB) transposon system has been shown to mediate new gene sequence integration resulting in long-term expression. Here the effectiveness of hyperactive SB100X transposase was tested, and we found that hydrodynamic co-delivery of a firefly luciferase transposon (pT2/CaL) along with SB100X transposase (pCMV-SB100X) resulted in remarkably sustained, high levels of luciferase expression. However, after 4 weeks there was a rapid, animal-by-animal loss of luciferase expression that was not observed in immunodeficient mice. We hypothesized that this sustained, high-level luciferase expression achieved using the SB100X transposase elicits an immune response in pT2/CaL co-administered mice, which was supported by the rapid loss of luciferase expression upon challenge of previously treated animals and in naive animals adoptively transferred with splenocytes from previously treated animals. Specificity of the immune response to luciferase was demonstrated by increased cytokine expression in splenocytes after exposure to luciferase peptide in parallel with MHC I-luciferase peptide tetramer binding. This anti-luciferase immune response observed following continuous, high-level luciferase expression in vivo clearly impacts its use as an in vivo reporter. As both an immunogen and an extremely sensitive reporter, luciferase is also a useful model system for the study of immune responses following in vivo gene transfer and expression.

AB - The Sleeping Beauty (SB) transposon system has been shown to mediate new gene sequence integration resulting in long-term expression. Here the effectiveness of hyperactive SB100X transposase was tested, and we found that hydrodynamic co-delivery of a firefly luciferase transposon (pT2/CaL) along with SB100X transposase (pCMV-SB100X) resulted in remarkably sustained, high levels of luciferase expression. However, after 4 weeks there was a rapid, animal-by-animal loss of luciferase expression that was not observed in immunodeficient mice. We hypothesized that this sustained, high-level luciferase expression achieved using the SB100X transposase elicits an immune response in pT2/CaL co-administered mice, which was supported by the rapid loss of luciferase expression upon challenge of previously treated animals and in naive animals adoptively transferred with splenocytes from previously treated animals. Specificity of the immune response to luciferase was demonstrated by increased cytokine expression in splenocytes after exposure to luciferase peptide in parallel with MHC I-luciferase peptide tetramer binding. This anti-luciferase immune response observed following continuous, high-level luciferase expression in vivo clearly impacts its use as an in vivo reporter. As both an immunogen and an extremely sensitive reporter, luciferase is also a useful model system for the study of immune responses following in vivo gene transfer and expression.

UR - http://www.scopus.com/inward/record.url?scp=84910677465&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84910677465&partnerID=8YFLogxK

U2 - 10.1089/hum.2014.048

DO - 10.1089/hum.2014.048

M3 - Article

VL - 25

SP - 955

EP - 965

JO - Human Gene Therapy

JF - Human Gene Therapy

SN - 1043-0342

IS - 11

ER -