Encapsulated cells producing retroviral vectors for in vivo gene transfer

Robert M. Saller, Stefano Indraccolo, Vincenzo Coppola, Giovanni Esposito, Jan Stange, Steffen Mitzner, Alberto Amadori, Brian Salmons, Walter H. Günzburg

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Background: Because gene therapy of the future will primarily take an in vivo approach, a number of problems associated with its current implementation exist. Currently, repeated delivery of a vector in vivo is necessary to ensure adequate transfer of the therapeutic gene. This may lead to the development of an immune response against the vector, thus interfering with gene delivery. To circumvent this problem, retroviral vector packaging cells that permanently produce recombinant retroviral vector particles have been encapsulated. Methods: Vector (pBAG)-producing amphotropic cells were encapsulated in beads composed of polymerized cellulose sulphate. These capsules were analysed in vitro for expression of the vector construct using X-gal staining, as well as for the release of particles by performing RT-PCR from culture supernatant. infectivity studies were performed in vitro and in vivo. The latter was assayed using histological sections of the microcapsule and the surrounding area stained for β-galactosidase activity and by RT-PCR. Results: In culture, the virus-producing cells inside the capsules remained viable and released virus into the culture medium for at least 6 weeks. To test whether these capsules, upon implantation into mice, also release vector virions that infect the surrounding cells, two different models were used. In the first, capsules were implanted in the fat pad of the mammary gland of Balb/c mice. The capsules were well tolerated for at least 6 weeks and a self-limiting inflammatory reaction without any other gross immune response was observed during this period. Furthermore, the virus-producing cells remained viable. In the second model, SCID mice were immunologically reconstituted by subcutaneous implantation of thymus lobes from MHC-identical Balb/c newborn mice and gene transfer into lymphoid cells was achieved by retroviral vectors released by co-implanted capsules. Conclusion: The implantation of such capsules containing cells that continually produce retroviral vector particles may be of use for in vivo gene therapy strategies. The data presented demonstrate the feasibility of the concept.

Original languageEnglish
Pages (from-to)150-160
Number of pages11
JournalJournal of Gene Medicine
Volume4
Issue number2
DOIs
Publication statusPublished - Mar 2002

Fingerprint

Capsules
Genes
Viruses
Genetic Therapy
Galactosidases
Polymerase Chain Reaction
SCID Mice
Product Packaging
Human Mammary Glands
Virion
Thymus Gland
Culture Media
Adipose Tissue
Lymphocytes
Staining and Labeling

Keywords

  • Cellulose sulphate
  • Encapsulation
  • In vivo delivery
  • Retroviral vectors

ASJC Scopus subject areas

  • Genetics

Cite this

Saller, R. M., Indraccolo, S., Coppola, V., Esposito, G., Stange, J., Mitzner, S., ... Günzburg, W. H. (2002). Encapsulated cells producing retroviral vectors for in vivo gene transfer. Journal of Gene Medicine, 4(2), 150-160. https://doi.org/10.1002/jgm.257

Encapsulated cells producing retroviral vectors for in vivo gene transfer. / Saller, Robert M.; Indraccolo, Stefano; Coppola, Vincenzo; Esposito, Giovanni; Stange, Jan; Mitzner, Steffen; Amadori, Alberto; Salmons, Brian; Günzburg, Walter H.

In: Journal of Gene Medicine, Vol. 4, No. 2, 03.2002, p. 150-160.

Research output: Contribution to journalArticle

Saller, RM, Indraccolo, S, Coppola, V, Esposito, G, Stange, J, Mitzner, S, Amadori, A, Salmons, B & Günzburg, WH 2002, 'Encapsulated cells producing retroviral vectors for in vivo gene transfer', Journal of Gene Medicine, vol. 4, no. 2, pp. 150-160. https://doi.org/10.1002/jgm.257
Saller, Robert M. ; Indraccolo, Stefano ; Coppola, Vincenzo ; Esposito, Giovanni ; Stange, Jan ; Mitzner, Steffen ; Amadori, Alberto ; Salmons, Brian ; Günzburg, Walter H. / Encapsulated cells producing retroviral vectors for in vivo gene transfer. In: Journal of Gene Medicine. 2002 ; Vol. 4, No. 2. pp. 150-160.
@article{ba447f1017544fc1987d89573e5b9be0,
title = "Encapsulated cells producing retroviral vectors for in vivo gene transfer",
abstract = "Background: Because gene therapy of the future will primarily take an in vivo approach, a number of problems associated with its current implementation exist. Currently, repeated delivery of a vector in vivo is necessary to ensure adequate transfer of the therapeutic gene. This may lead to the development of an immune response against the vector, thus interfering with gene delivery. To circumvent this problem, retroviral vector packaging cells that permanently produce recombinant retroviral vector particles have been encapsulated. Methods: Vector (pBAG)-producing amphotropic cells were encapsulated in beads composed of polymerized cellulose sulphate. These capsules were analysed in vitro for expression of the vector construct using X-gal staining, as well as for the release of particles by performing RT-PCR from culture supernatant. infectivity studies were performed in vitro and in vivo. The latter was assayed using histological sections of the microcapsule and the surrounding area stained for β-galactosidase activity and by RT-PCR. Results: In culture, the virus-producing cells inside the capsules remained viable and released virus into the culture medium for at least 6 weeks. To test whether these capsules, upon implantation into mice, also release vector virions that infect the surrounding cells, two different models were used. In the first, capsules were implanted in the fat pad of the mammary gland of Balb/c mice. The capsules were well tolerated for at least 6 weeks and a self-limiting inflammatory reaction without any other gross immune response was observed during this period. Furthermore, the virus-producing cells remained viable. In the second model, SCID mice were immunologically reconstituted by subcutaneous implantation of thymus lobes from MHC-identical Balb/c newborn mice and gene transfer into lymphoid cells was achieved by retroviral vectors released by co-implanted capsules. Conclusion: The implantation of such capsules containing cells that continually produce retroviral vector particles may be of use for in vivo gene therapy strategies. The data presented demonstrate the feasibility of the concept.",
keywords = "Cellulose sulphate, Encapsulation, In vivo delivery, Retroviral vectors",
author = "Saller, {Robert M.} and Stefano Indraccolo and Vincenzo Coppola and Giovanni Esposito and Jan Stange and Steffen Mitzner and Alberto Amadori and Brian Salmons and G{\"u}nzburg, {Walter H.}",
year = "2002",
month = "3",
doi = "10.1002/jgm.257",
language = "English",
volume = "4",
pages = "150--160",
journal = "Journal of Gene Medicine",
issn = "1099-498X",
publisher = "John Wiley and Sons Ltd",
number = "2",

}

TY - JOUR

T1 - Encapsulated cells producing retroviral vectors for in vivo gene transfer

AU - Saller, Robert M.

AU - Indraccolo, Stefano

AU - Coppola, Vincenzo

AU - Esposito, Giovanni

AU - Stange, Jan

AU - Mitzner, Steffen

AU - Amadori, Alberto

AU - Salmons, Brian

AU - Günzburg, Walter H.

PY - 2002/3

Y1 - 2002/3

N2 - Background: Because gene therapy of the future will primarily take an in vivo approach, a number of problems associated with its current implementation exist. Currently, repeated delivery of a vector in vivo is necessary to ensure adequate transfer of the therapeutic gene. This may lead to the development of an immune response against the vector, thus interfering with gene delivery. To circumvent this problem, retroviral vector packaging cells that permanently produce recombinant retroviral vector particles have been encapsulated. Methods: Vector (pBAG)-producing amphotropic cells were encapsulated in beads composed of polymerized cellulose sulphate. These capsules were analysed in vitro for expression of the vector construct using X-gal staining, as well as for the release of particles by performing RT-PCR from culture supernatant. infectivity studies were performed in vitro and in vivo. The latter was assayed using histological sections of the microcapsule and the surrounding area stained for β-galactosidase activity and by RT-PCR. Results: In culture, the virus-producing cells inside the capsules remained viable and released virus into the culture medium for at least 6 weeks. To test whether these capsules, upon implantation into mice, also release vector virions that infect the surrounding cells, two different models were used. In the first, capsules were implanted in the fat pad of the mammary gland of Balb/c mice. The capsules were well tolerated for at least 6 weeks and a self-limiting inflammatory reaction without any other gross immune response was observed during this period. Furthermore, the virus-producing cells remained viable. In the second model, SCID mice were immunologically reconstituted by subcutaneous implantation of thymus lobes from MHC-identical Balb/c newborn mice and gene transfer into lymphoid cells was achieved by retroviral vectors released by co-implanted capsules. Conclusion: The implantation of such capsules containing cells that continually produce retroviral vector particles may be of use for in vivo gene therapy strategies. The data presented demonstrate the feasibility of the concept.

AB - Background: Because gene therapy of the future will primarily take an in vivo approach, a number of problems associated with its current implementation exist. Currently, repeated delivery of a vector in vivo is necessary to ensure adequate transfer of the therapeutic gene. This may lead to the development of an immune response against the vector, thus interfering with gene delivery. To circumvent this problem, retroviral vector packaging cells that permanently produce recombinant retroviral vector particles have been encapsulated. Methods: Vector (pBAG)-producing amphotropic cells were encapsulated in beads composed of polymerized cellulose sulphate. These capsules were analysed in vitro for expression of the vector construct using X-gal staining, as well as for the release of particles by performing RT-PCR from culture supernatant. infectivity studies were performed in vitro and in vivo. The latter was assayed using histological sections of the microcapsule and the surrounding area stained for β-galactosidase activity and by RT-PCR. Results: In culture, the virus-producing cells inside the capsules remained viable and released virus into the culture medium for at least 6 weeks. To test whether these capsules, upon implantation into mice, also release vector virions that infect the surrounding cells, two different models were used. In the first, capsules were implanted in the fat pad of the mammary gland of Balb/c mice. The capsules were well tolerated for at least 6 weeks and a self-limiting inflammatory reaction without any other gross immune response was observed during this period. Furthermore, the virus-producing cells remained viable. In the second model, SCID mice were immunologically reconstituted by subcutaneous implantation of thymus lobes from MHC-identical Balb/c newborn mice and gene transfer into lymphoid cells was achieved by retroviral vectors released by co-implanted capsules. Conclusion: The implantation of such capsules containing cells that continually produce retroviral vector particles may be of use for in vivo gene therapy strategies. The data presented demonstrate the feasibility of the concept.

KW - Cellulose sulphate

KW - Encapsulation

KW - In vivo delivery

KW - Retroviral vectors

UR - http://www.scopus.com/inward/record.url?scp=0036518340&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036518340&partnerID=8YFLogxK

U2 - 10.1002/jgm.257

DO - 10.1002/jgm.257

M3 - Article

VL - 4

SP - 150

EP - 160

JO - Journal of Gene Medicine

JF - Journal of Gene Medicine

SN - 1099-498X

IS - 2

ER -