A dendritic cell-based assay for measuring memory T cells specific to dengue envelope proteins in human peripheral blood

Peifang Sun; Charmagne Beckett; Janine Danko; Timothy Burgess; Zhaodong Liang; Tadeusz Kochel; Kevin Porter

doi:10.1016/j.jviromet.2011.01.023

A dendritic cell-based assay for measuring memory T cells specific to dengue envelope proteins in human peripheral blood

Peifang Sun^*, Charmagne Beckett, Janine Danko, Timothy Burgess, Zhaodong Liang, Tadeusz Kochel, Kevin Porter

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

Dengue envelope (E) protein is a dominant immune inducer and E protein-based vaccines elicited partial to complete protection in non-human primates. To study the immunogenicity of these vaccines in humans, an enzyme linked immunospot (ELISPOT) assay for measuring interferon gamma (IFN-γ) production was developed. Cells from two subject groups, based on dengue-exposure, were selected for assay development. The unique feature of the IFN-γ ELISPOT assay is the utilization of dendritic cells pulsed with E proteins as antigen presenting cells. IFN-γ production, ranging from 53-513 spot forming units per million peripheral blood mononuclear cells (PBMCs), was observed in dengue-exposed subjects as compared to 0-45 IFN-γ spot forming units in dengue-unexposed subjects. Further, both CD4⁺ and CD8⁺ T cells, and cells bearing CD45RO memory marker, were the major sources of IFN-γ production. The assay allowed quantification of E-specific IFN-γ-secreting memory T cells in subjects 9 years after exposure to a live-attenuated virus vaccine and live-virus challenge. Results suggested that the dendritic cell-based IFN-γ assay is a useful tool for assessing immunological memory for clinical research.

Original language	English
Pages (from-to)	175-181
Number of pages	7
Journal	Journal of Virological Methods
Volume	173
Issue number	2
DOIs	https://doi.org/10.1016/j.jviromet.2011.01.023
State	Published - May 2011

Keywords

Dengue
Dengue E protein
Human T cell
Human dendritic cell
Interferon ELISPOT

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10.1016/j.jviromet.2011.01.023

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title = "A dendritic cell-based assay for measuring memory T cells specific to dengue envelope proteins in human peripheral blood",

abstract = "Dengue envelope (E) protein is a dominant immune inducer and E protein-based vaccines elicited partial to complete protection in non-human primates. To study the immunogenicity of these vaccines in humans, an enzyme linked immunospot (ELISPOT) assay for measuring interferon gamma (IFN-γ) production was developed. Cells from two subject groups, based on dengue-exposure, were selected for assay development. The unique feature of the IFN-γ ELISPOT assay is the utilization of dendritic cells pulsed with E proteins as antigen presenting cells. IFN-γ production, ranging from 53-513 spot forming units per million peripheral blood mononuclear cells (PBMCs), was observed in dengue-exposed subjects as compared to 0-45 IFN-γ spot forming units in dengue-unexposed subjects. Further, both CD4+ and CD8+ T cells, and cells bearing CD45RO memory marker, were the major sources of IFN-γ production. The assay allowed quantification of E-specific IFN-γ-secreting memory T cells in subjects 9 years after exposure to a live-attenuated virus vaccine and live-virus challenge. Results suggested that the dendritic cell-based IFN-γ assay is a useful tool for assessing immunological memory for clinical research.",

keywords = "Dengue, Dengue E protein, Human T cell, Human dendritic cell, Interferon ELISPOT",

author = "Peifang Sun and Charmagne Beckett and Janine Danko and Timothy Burgess and Zhaodong Liang and Tadeusz Kochel and Kevin Porter",

note = "Funding Information: This work was supported by U.S. Navy work unit number 6000.RAD1.S.A0312 . ",

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AU - Sun, Peifang

AU - Beckett, Charmagne

AU - Danko, Janine

AU - Burgess, Timothy

AU - Liang, Zhaodong

AU - Kochel, Tadeusz

AU - Porter, Kevin

N1 - Funding Information: This work was supported by U.S. Navy work unit number 6000.RAD1.S.A0312 .

PY - 2011/5

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N2 - Dengue envelope (E) protein is a dominant immune inducer and E protein-based vaccines elicited partial to complete protection in non-human primates. To study the immunogenicity of these vaccines in humans, an enzyme linked immunospot (ELISPOT) assay for measuring interferon gamma (IFN-γ) production was developed. Cells from two subject groups, based on dengue-exposure, were selected for assay development. The unique feature of the IFN-γ ELISPOT assay is the utilization of dendritic cells pulsed with E proteins as antigen presenting cells. IFN-γ production, ranging from 53-513 spot forming units per million peripheral blood mononuclear cells (PBMCs), was observed in dengue-exposed subjects as compared to 0-45 IFN-γ spot forming units in dengue-unexposed subjects. Further, both CD4+ and CD8+ T cells, and cells bearing CD45RO memory marker, were the major sources of IFN-γ production. The assay allowed quantification of E-specific IFN-γ-secreting memory T cells in subjects 9 years after exposure to a live-attenuated virus vaccine and live-virus challenge. Results suggested that the dendritic cell-based IFN-γ assay is a useful tool for assessing immunological memory for clinical research.

AB - Dengue envelope (E) protein is a dominant immune inducer and E protein-based vaccines elicited partial to complete protection in non-human primates. To study the immunogenicity of these vaccines in humans, an enzyme linked immunospot (ELISPOT) assay for measuring interferon gamma (IFN-γ) production was developed. Cells from two subject groups, based on dengue-exposure, were selected for assay development. The unique feature of the IFN-γ ELISPOT assay is the utilization of dendritic cells pulsed with E proteins as antigen presenting cells. IFN-γ production, ranging from 53-513 spot forming units per million peripheral blood mononuclear cells (PBMCs), was observed in dengue-exposed subjects as compared to 0-45 IFN-γ spot forming units in dengue-unexposed subjects. Further, both CD4+ and CD8+ T cells, and cells bearing CD45RO memory marker, were the major sources of IFN-γ production. The assay allowed quantification of E-specific IFN-γ-secreting memory T cells in subjects 9 years after exposure to a live-attenuated virus vaccine and live-virus challenge. Results suggested that the dendritic cell-based IFN-γ assay is a useful tool for assessing immunological memory for clinical research.

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A dendritic cell-based assay for measuring memory T cells specific to dengue envelope proteins in human peripheral blood

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Keywords

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