Construction and expression in tumor cells of a recombinant vaccinia virus encoding human interleukin-1β

Gary R. Peplinski, Kangla Tsung, Eric Whitman, Jennifer B. Meko, Jeffrey A. Norton

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Background: Human interleukin-1β (hIL-1β) injected intratumorally has demonstrated growth inhibition of transplanted subcutaneous tumors in mice, regression of metastatic lesions, resistance to tumor rechallenge, and increased survival. Vaccinia virus (VV) can be genetically engineered to produce cytokines and may be an effective vector for gene therapy of cancer. This study was designed to (a) construct a VV expressing hIL-1β, (b) assess tumor cell infection in vitro with this construct, (c) measure hIL-1β production, and (d) assess the bioactivity of the secreted cytokine. Methods: The hIL-1β gene was amplified from a plasmid clone using polymerase chain reaction (PCR) and then cloned into a homologous recombination (HR) and expression vector, which was used to insert the hIL-1β gene into the VV genome. Selection of the recombinant VV (vMJ601hIL-1β) was based on inactivation of viral TK and expression of β-galactosidase. vMJ601hIL-1β infectivity and cytokine production was assessed by infecting tumor cell lines and analyzing culture supernatants for hIL-1β. Bioactivity of the hIL-1β produced was demonstrated using an IL-1 dependent T helper cell line. Results: The hIL-1β gene was successfully cloned into the VV genome by HR, which was confirmed by PCR. vMJ601hIL-1β efficiently infected tumor cells, as shown by increased hIL-1β secretion (0 to >500 ng/ml) and morphologic evidence of viral cytopathic effect. vMJ601hIL-1β-infected cells secreted large amounts of hIL-1β (mean 772 ng/106 cells/24 h). The secreted hIL-1β was bioactive (mean bioactivity 6.8×108 U/mg of hIL-1β). Conclusions: (a) hIL-1β can be cloned into VV, (b) vMJ601hIL-1β retains its infectivity, (c) a large amount of hIL-1β is secreted, and (d) the secreted hIL-1β is bioactive. Recombinant VV may allow in situ cytokine gene delivery and expression in established tumors.

Original languageEnglish (US)
Pages (from-to)151-159
Number of pages9
JournalAnnals of Surgical Oncology
Volume2
Issue number2
DOIs
StatePublished - Mar 1 1995

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Vaccinia virus
Interleukin-1
Neoplasms
Cytokines
Homologous Recombination
Viral Cytopathogenic Effect
Galactosidases
Genome
Virus Inactivation
Genes
Polymerase Chain Reaction
Tumor Cell Line
Genetic Therapy

All Science Journal Classification (ASJC) codes

  • Surgery
  • Oncology

Cite this

Peplinski, Gary R. ; Tsung, Kangla ; Whitman, Eric ; Meko, Jennifer B. ; Norton, Jeffrey A. / Construction and expression in tumor cells of a recombinant vaccinia virus encoding human interleukin-1β. In: Annals of Surgical Oncology. 1995 ; Vol. 2, No. 2. pp. 151-159.
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title = "Construction and expression in tumor cells of a recombinant vaccinia virus encoding human interleukin-1β",
abstract = "Background: Human interleukin-1β (hIL-1β) injected intratumorally has demonstrated growth inhibition of transplanted subcutaneous tumors in mice, regression of metastatic lesions, resistance to tumor rechallenge, and increased survival. Vaccinia virus (VV) can be genetically engineered to produce cytokines and may be an effective vector for gene therapy of cancer. This study was designed to (a) construct a VV expressing hIL-1β, (b) assess tumor cell infection in vitro with this construct, (c) measure hIL-1β production, and (d) assess the bioactivity of the secreted cytokine. Methods: The hIL-1β gene was amplified from a plasmid clone using polymerase chain reaction (PCR) and then cloned into a homologous recombination (HR) and expression vector, which was used to insert the hIL-1β gene into the VV genome. Selection of the recombinant VV (vMJ601hIL-1β) was based on inactivation of viral TK and expression of β-galactosidase. vMJ601hIL-1β infectivity and cytokine production was assessed by infecting tumor cell lines and analyzing culture supernatants for hIL-1β. Bioactivity of the hIL-1β produced was demonstrated using an IL-1 dependent T helper cell line. Results: The hIL-1β gene was successfully cloned into the VV genome by HR, which was confirmed by PCR. vMJ601hIL-1β efficiently infected tumor cells, as shown by increased hIL-1β secretion (0 to >500 ng/ml) and morphologic evidence of viral cytopathic effect. vMJ601hIL-1β-infected cells secreted large amounts of hIL-1β (mean 772 ng/106 cells/24 h). The secreted hIL-1β was bioactive (mean bioactivity 6.8×108 U/mg of hIL-1β). Conclusions: (a) hIL-1β can be cloned into VV, (b) vMJ601hIL-1β retains its infectivity, (c) a large amount of hIL-1β is secreted, and (d) the secreted hIL-1β is bioactive. Recombinant VV may allow in situ cytokine gene delivery and expression in established tumors.",
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Construction and expression in tumor cells of a recombinant vaccinia virus encoding human interleukin-1β. / Peplinski, Gary R.; Tsung, Kangla; Whitman, Eric; Meko, Jennifer B.; Norton, Jeffrey A.

In: Annals of Surgical Oncology, Vol. 2, No. 2, 01.03.1995, p. 151-159.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Construction and expression in tumor cells of a recombinant vaccinia virus encoding human interleukin-1β

AU - Peplinski, Gary R.

AU - Tsung, Kangla

AU - Whitman, Eric

AU - Meko, Jennifer B.

AU - Norton, Jeffrey A.

PY - 1995/3/1

Y1 - 1995/3/1

N2 - Background: Human interleukin-1β (hIL-1β) injected intratumorally has demonstrated growth inhibition of transplanted subcutaneous tumors in mice, regression of metastatic lesions, resistance to tumor rechallenge, and increased survival. Vaccinia virus (VV) can be genetically engineered to produce cytokines and may be an effective vector for gene therapy of cancer. This study was designed to (a) construct a VV expressing hIL-1β, (b) assess tumor cell infection in vitro with this construct, (c) measure hIL-1β production, and (d) assess the bioactivity of the secreted cytokine. Methods: The hIL-1β gene was amplified from a plasmid clone using polymerase chain reaction (PCR) and then cloned into a homologous recombination (HR) and expression vector, which was used to insert the hIL-1β gene into the VV genome. Selection of the recombinant VV (vMJ601hIL-1β) was based on inactivation of viral TK and expression of β-galactosidase. vMJ601hIL-1β infectivity and cytokine production was assessed by infecting tumor cell lines and analyzing culture supernatants for hIL-1β. Bioactivity of the hIL-1β produced was demonstrated using an IL-1 dependent T helper cell line. Results: The hIL-1β gene was successfully cloned into the VV genome by HR, which was confirmed by PCR. vMJ601hIL-1β efficiently infected tumor cells, as shown by increased hIL-1β secretion (0 to >500 ng/ml) and morphologic evidence of viral cytopathic effect. vMJ601hIL-1β-infected cells secreted large amounts of hIL-1β (mean 772 ng/106 cells/24 h). The secreted hIL-1β was bioactive (mean bioactivity 6.8×108 U/mg of hIL-1β). Conclusions: (a) hIL-1β can be cloned into VV, (b) vMJ601hIL-1β retains its infectivity, (c) a large amount of hIL-1β is secreted, and (d) the secreted hIL-1β is bioactive. Recombinant VV may allow in situ cytokine gene delivery and expression in established tumors.

AB - Background: Human interleukin-1β (hIL-1β) injected intratumorally has demonstrated growth inhibition of transplanted subcutaneous tumors in mice, regression of metastatic lesions, resistance to tumor rechallenge, and increased survival. Vaccinia virus (VV) can be genetically engineered to produce cytokines and may be an effective vector for gene therapy of cancer. This study was designed to (a) construct a VV expressing hIL-1β, (b) assess tumor cell infection in vitro with this construct, (c) measure hIL-1β production, and (d) assess the bioactivity of the secreted cytokine. Methods: The hIL-1β gene was amplified from a plasmid clone using polymerase chain reaction (PCR) and then cloned into a homologous recombination (HR) and expression vector, which was used to insert the hIL-1β gene into the VV genome. Selection of the recombinant VV (vMJ601hIL-1β) was based on inactivation of viral TK and expression of β-galactosidase. vMJ601hIL-1β infectivity and cytokine production was assessed by infecting tumor cell lines and analyzing culture supernatants for hIL-1β. Bioactivity of the hIL-1β produced was demonstrated using an IL-1 dependent T helper cell line. Results: The hIL-1β gene was successfully cloned into the VV genome by HR, which was confirmed by PCR. vMJ601hIL-1β efficiently infected tumor cells, as shown by increased hIL-1β secretion (0 to >500 ng/ml) and morphologic evidence of viral cytopathic effect. vMJ601hIL-1β-infected cells secreted large amounts of hIL-1β (mean 772 ng/106 cells/24 h). The secreted hIL-1β was bioactive (mean bioactivity 6.8×108 U/mg of hIL-1β). Conclusions: (a) hIL-1β can be cloned into VV, (b) vMJ601hIL-1β retains its infectivity, (c) a large amount of hIL-1β is secreted, and (d) the secreted hIL-1β is bioactive. Recombinant VV may allow in situ cytokine gene delivery and expression in established tumors.

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