Drug-inducible, dendritic cell-based genetic immunization

Laura Timares, Karim Mahmoud Safer, Baoxi Qu, Akira Takashima, Stephen Johnston

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Determining the mechanism of Ag loading of Langerhans cells (LC) for genetic immunization (GI) is complicated by the inability to distinguish between the response generated by direct transfection of LC from that due to exogenous uptake. To unravel this mechanism, we examined the impact of gene gun treatment on LC with respect to their activation and migration from skin, transgene expression, and ability to initiate humoral and cellular immune responses upon transfer to naive mice. To assess responses generated by direct LC transfection, an RU486-inducible expression system was used as a GI vector. In vitro skin organ cultures were developed from gene gun immunized mouse ear specimens to obtain LC. Gene gun treatment markedly augmented (3-fold) LC migration from ear skin, and these LC expressed the transgene at RNA and protein levels. Transfer of 2 × 105 migratory cells resulted in identical cellular responses to, but 10-fold lower humoral responses than, standard GI. Using an RU486-inducible system, we were able to measure responses generated by directly transfected LC. Our results indicate that direct transfection is a predominant pathway for LC Ag loading. The ability to regulate transgene expression with inducible DC-based vaccines demonstrates a new level of immunological control.

Original languageEnglish (US)
Pages (from-to)5483-5490
Number of pages8
JournalJournal of Immunology
Volume170
Issue number11
StatePublished - Jun 1 2003
Externally publishedYes

Fingerprint

Langerhans Cells
Dendritic Cells
Immunization
Pharmaceutical Preparations
Firearms
Transgenes
Transfection
Skin
Ear
Genes
Genetic Vectors
Organ Culture Techniques
Humoral Immunity
Cellular Immunity
Cell Movement
Vaccines
RNA

ASJC Scopus subject areas

  • Immunology

Cite this

Timares, L., Mahmoud Safer, K., Qu, B., Takashima, A., & Johnston, S. (2003). Drug-inducible, dendritic cell-based genetic immunization. Journal of Immunology, 170(11), 5483-5490.

Drug-inducible, dendritic cell-based genetic immunization. / Timares, Laura; Mahmoud Safer, Karim; Qu, Baoxi; Takashima, Akira; Johnston, Stephen.

In: Journal of Immunology, Vol. 170, No. 11, 01.06.2003, p. 5483-5490.

Research output: Contribution to journalArticle

Timares, L, Mahmoud Safer, K, Qu, B, Takashima, A & Johnston, S 2003, 'Drug-inducible, dendritic cell-based genetic immunization', Journal of Immunology, vol. 170, no. 11, pp. 5483-5490.
Timares L, Mahmoud Safer K, Qu B, Takashima A, Johnston S. Drug-inducible, dendritic cell-based genetic immunization. Journal of Immunology. 2003 Jun 1;170(11):5483-5490.
Timares, Laura ; Mahmoud Safer, Karim ; Qu, Baoxi ; Takashima, Akira ; Johnston, Stephen. / Drug-inducible, dendritic cell-based genetic immunization. In: Journal of Immunology. 2003 ; Vol. 170, No. 11. pp. 5483-5490.
@article{85f45256820d4ed59ddadfe01b751cf1,
title = "Drug-inducible, dendritic cell-based genetic immunization",
abstract = "Determining the mechanism of Ag loading of Langerhans cells (LC) for genetic immunization (GI) is complicated by the inability to distinguish between the response generated by direct transfection of LC from that due to exogenous uptake. To unravel this mechanism, we examined the impact of gene gun treatment on LC with respect to their activation and migration from skin, transgene expression, and ability to initiate humoral and cellular immune responses upon transfer to naive mice. To assess responses generated by direct LC transfection, an RU486-inducible expression system was used as a GI vector. In vitro skin organ cultures were developed from gene gun immunized mouse ear specimens to obtain LC. Gene gun treatment markedly augmented (3-fold) LC migration from ear skin, and these LC expressed the transgene at RNA and protein levels. Transfer of 2 × 105 migratory cells resulted in identical cellular responses to, but 10-fold lower humoral responses than, standard GI. Using an RU486-inducible system, we were able to measure responses generated by directly transfected LC. Our results indicate that direct transfection is a predominant pathway for LC Ag loading. The ability to regulate transgene expression with inducible DC-based vaccines demonstrates a new level of immunological control.",
author = "Laura Timares and {Mahmoud Safer}, Karim and Baoxi Qu and Akira Takashima and Stephen Johnston",
year = "2003",
month = "6",
day = "1",
language = "English (US)",
volume = "170",
pages = "5483--5490",
journal = "Journal of Immunology",
issn = "0022-1767",
publisher = "American Association of Immunologists",
number = "11",

}

TY - JOUR

T1 - Drug-inducible, dendritic cell-based genetic immunization

AU - Timares, Laura

AU - Mahmoud Safer, Karim

AU - Qu, Baoxi

AU - Takashima, Akira

AU - Johnston, Stephen

PY - 2003/6/1

Y1 - 2003/6/1

N2 - Determining the mechanism of Ag loading of Langerhans cells (LC) for genetic immunization (GI) is complicated by the inability to distinguish between the response generated by direct transfection of LC from that due to exogenous uptake. To unravel this mechanism, we examined the impact of gene gun treatment on LC with respect to their activation and migration from skin, transgene expression, and ability to initiate humoral and cellular immune responses upon transfer to naive mice. To assess responses generated by direct LC transfection, an RU486-inducible expression system was used as a GI vector. In vitro skin organ cultures were developed from gene gun immunized mouse ear specimens to obtain LC. Gene gun treatment markedly augmented (3-fold) LC migration from ear skin, and these LC expressed the transgene at RNA and protein levels. Transfer of 2 × 105 migratory cells resulted in identical cellular responses to, but 10-fold lower humoral responses than, standard GI. Using an RU486-inducible system, we were able to measure responses generated by directly transfected LC. Our results indicate that direct transfection is a predominant pathway for LC Ag loading. The ability to regulate transgene expression with inducible DC-based vaccines demonstrates a new level of immunological control.

AB - Determining the mechanism of Ag loading of Langerhans cells (LC) for genetic immunization (GI) is complicated by the inability to distinguish between the response generated by direct transfection of LC from that due to exogenous uptake. To unravel this mechanism, we examined the impact of gene gun treatment on LC with respect to their activation and migration from skin, transgene expression, and ability to initiate humoral and cellular immune responses upon transfer to naive mice. To assess responses generated by direct LC transfection, an RU486-inducible expression system was used as a GI vector. In vitro skin organ cultures were developed from gene gun immunized mouse ear specimens to obtain LC. Gene gun treatment markedly augmented (3-fold) LC migration from ear skin, and these LC expressed the transgene at RNA and protein levels. Transfer of 2 × 105 migratory cells resulted in identical cellular responses to, but 10-fold lower humoral responses than, standard GI. Using an RU486-inducible system, we were able to measure responses generated by directly transfected LC. Our results indicate that direct transfection is a predominant pathway for LC Ag loading. The ability to regulate transgene expression with inducible DC-based vaccines demonstrates a new level of immunological control.

UR - http://www.scopus.com/inward/record.url?scp=0037514400&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037514400&partnerID=8YFLogxK

M3 - Article

C2 - 12759425

AN - SCOPUS:0037514400

VL - 170

SP - 5483

EP - 5490

JO - Journal of Immunology

JF - Journal of Immunology

SN - 0022-1767

IS - 11

ER -