Abstract
The enzyme-linked immune spot (ELISPOT) assay is receiving increased attention as a means for quantifying antigen-specific CD8 T-cell responses in rhesus macaques. Further improving the sensitivity of this assay could aid in the evaluation of vaccine candidates and/or immune therapeutic candidates. Interleukin (IL)-15 has been demonstrated to stimulate expansion of human immunodeficiency virus (HIV)-specific cytotoxic T lymphocytes (CTL) and to regulate homeostatic proliferation of CD8+ memory cells. We evaluated the in vitro effect of IL-15 to increase the detection of interferon-γ (IFN-γ) production by antigen-specific stimulated lymphocytes from a group of rhesus macaques exposed to simian-human immunodeficiency virus (SHIV) and a second group infected with SIVmac251, before and after antiretroviral treatment (ART). Results from these studies demonstrate that the presence of IL-15 during stimulation in a peptide-based ELISPOT assay greatly enhanced IFN-γ production in both SHIV and simian immunodeficiency virus (SIV)-infected macaques. IFN-γ production was mainly mediated by CD8 lymphocytes. The optimal concentrations of IL-15 that give enhancement of IFN-γ production to specific antigen, without a significant increase in the spontaneous IFN-γ release, ranged from 0.5 to 2.5 ng/ml. The mean number of IFN-γ spots was increased 3.1- to 3.6-fold in response to SIV gag or HIV env peptide pools, respectively, in peripheral blood mononuclear cells (PBMC) from SHIV-infected macaques. Similarly, in SIV-infected macaques, IL-15 increased the mean number of IFN-γ spots 2.7-fold in response to both SIV gag and env peptide pools. In samples obtained after ART in the same macaques, the increase factor was 2.5 for SIV gag and 1.8 for the env peptide pools. Thus, the sensitivity of the ELISPOT assay can be enhanced by addition of IL-15. This modified assay will be useful for detection of low frequencies of IFN-γ producing cells in rhesus macaques.
| Original language | English |
|---|---|
| Pages (from-to) | 55-67 |
| Number of pages | 13 |
| Journal | Journal of Immunological Methods |
| Volume | 279 |
| Issue number | 1-2 |
| DOIs | |
| Publication status | Published - Aug 2003 |
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Keywords
- ELISPOT
- IL-15
- Interferon-γ
- Rhesus macaques
ASJC Scopus subject areas
- Biotechnology
- Immunology
Cite this
Use of interleukin 15 to enhance interferon-γ production by antigen-specific stimulated lymphocytes from rhesus macaques. / Calarota, Sandra A.; Otero, Miguel; Hermanstayne, Keith; Lewis, Mark; Rosati, Margherita; Felber, Barbara K.; Pavlakis, George N.; Boyer, Jean D.; Weiner, David B.
In: Journal of Immunological Methods, Vol. 279, No. 1-2, 08.2003, p. 55-67.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Use of interleukin 15 to enhance interferon-γ production by antigen-specific stimulated lymphocytes from rhesus macaques
AU - Calarota, Sandra A.
AU - Otero, Miguel
AU - Hermanstayne, Keith
AU - Lewis, Mark
AU - Rosati, Margherita
AU - Felber, Barbara K.
AU - Pavlakis, George N.
AU - Boyer, Jean D.
AU - Weiner, David B.
PY - 2003/8
Y1 - 2003/8
N2 - The enzyme-linked immune spot (ELISPOT) assay is receiving increased attention as a means for quantifying antigen-specific CD8 T-cell responses in rhesus macaques. Further improving the sensitivity of this assay could aid in the evaluation of vaccine candidates and/or immune therapeutic candidates. Interleukin (IL)-15 has been demonstrated to stimulate expansion of human immunodeficiency virus (HIV)-specific cytotoxic T lymphocytes (CTL) and to regulate homeostatic proliferation of CD8+ memory cells. We evaluated the in vitro effect of IL-15 to increase the detection of interferon-γ (IFN-γ) production by antigen-specific stimulated lymphocytes from a group of rhesus macaques exposed to simian-human immunodeficiency virus (SHIV) and a second group infected with SIVmac251, before and after antiretroviral treatment (ART). Results from these studies demonstrate that the presence of IL-15 during stimulation in a peptide-based ELISPOT assay greatly enhanced IFN-γ production in both SHIV and simian immunodeficiency virus (SIV)-infected macaques. IFN-γ production was mainly mediated by CD8 lymphocytes. The optimal concentrations of IL-15 that give enhancement of IFN-γ production to specific antigen, without a significant increase in the spontaneous IFN-γ release, ranged from 0.5 to 2.5 ng/ml. The mean number of IFN-γ spots was increased 3.1- to 3.6-fold in response to SIV gag or HIV env peptide pools, respectively, in peripheral blood mononuclear cells (PBMC) from SHIV-infected macaques. Similarly, in SIV-infected macaques, IL-15 increased the mean number of IFN-γ spots 2.7-fold in response to both SIV gag and env peptide pools. In samples obtained after ART in the same macaques, the increase factor was 2.5 for SIV gag and 1.8 for the env peptide pools. Thus, the sensitivity of the ELISPOT assay can be enhanced by addition of IL-15. This modified assay will be useful for detection of low frequencies of IFN-γ producing cells in rhesus macaques.
AB - The enzyme-linked immune spot (ELISPOT) assay is receiving increased attention as a means for quantifying antigen-specific CD8 T-cell responses in rhesus macaques. Further improving the sensitivity of this assay could aid in the evaluation of vaccine candidates and/or immune therapeutic candidates. Interleukin (IL)-15 has been demonstrated to stimulate expansion of human immunodeficiency virus (HIV)-specific cytotoxic T lymphocytes (CTL) and to regulate homeostatic proliferation of CD8+ memory cells. We evaluated the in vitro effect of IL-15 to increase the detection of interferon-γ (IFN-γ) production by antigen-specific stimulated lymphocytes from a group of rhesus macaques exposed to simian-human immunodeficiency virus (SHIV) and a second group infected with SIVmac251, before and after antiretroviral treatment (ART). Results from these studies demonstrate that the presence of IL-15 during stimulation in a peptide-based ELISPOT assay greatly enhanced IFN-γ production in both SHIV and simian immunodeficiency virus (SIV)-infected macaques. IFN-γ production was mainly mediated by CD8 lymphocytes. The optimal concentrations of IL-15 that give enhancement of IFN-γ production to specific antigen, without a significant increase in the spontaneous IFN-γ release, ranged from 0.5 to 2.5 ng/ml. The mean number of IFN-γ spots was increased 3.1- to 3.6-fold in response to SIV gag or HIV env peptide pools, respectively, in peripheral blood mononuclear cells (PBMC) from SHIV-infected macaques. Similarly, in SIV-infected macaques, IL-15 increased the mean number of IFN-γ spots 2.7-fold in response to both SIV gag and env peptide pools. In samples obtained after ART in the same macaques, the increase factor was 2.5 for SIV gag and 1.8 for the env peptide pools. Thus, the sensitivity of the ELISPOT assay can be enhanced by addition of IL-15. This modified assay will be useful for detection of low frequencies of IFN-γ producing cells in rhesus macaques.
KW - ELISPOT
KW - IL-15
KW - Interferon-γ
KW - Rhesus macaques
UR - http://www.scopus.com/inward/record.url?scp=0142061097&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0142061097&partnerID=8YFLogxK
U2 - 10.1016/S0022-1759(03)00246-1
DO - 10.1016/S0022-1759(03)00246-1
M3 - Article
C2 - 12969547
AN - SCOPUS:0142061097
VL - 279
SP - 55
EP - 67
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1-2
ER -