TY - JOUR
T1 - A dendritic cell-based assay for measuring memory T cells specific to dengue envelope proteins in human peripheral blood
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
Y1 - 2011/5
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.
KW - Dengue
KW - Dengue E protein
KW - Human T cell
KW - Human dendritic cell
KW - Interferon ELISPOT
UR - http://www.scopus.com/inward/record.url?scp=79954601348&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2011.01.023
DO - 10.1016/j.jviromet.2011.01.023
M3 - Article
C2 - 21315764
AN - SCOPUS:79954601348
SN - 0166-0934
VL - 173
SP - 175
EP - 181
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 2
ER -