Reader-free ELISPOT assay for immuno-monitoring in peptide-based cancer vaccine immunotherapy.
ELISPOT
cancer vaccine
immuno-monitoring
immunotherapy
Journal
Biomedical reports
ISSN: 2049-9434
Titre abrégé: Biomed Rep
Pays: England
ID NLM: 101613227
Informations de publication
Date de publication:
May 2020
May 2020
Historique:
received:
24
06
2019
accepted:
03
01
2020
entrez:
8
4
2020
pubmed:
8
4
2020
medline:
8
4
2020
Statut:
ppublish
Résumé
Cancer vaccine immunotherapy is a therapy that induces cellular immune responses against a target molecule to elicit clinical anti-tumor effects. These cellular immune responses against the target molecule are monitored to evaluate whether the antigen-specific cellular immune responses are induced and maintained during the vaccination period. Enzyme-linked immunospot (ELISPOT) assay is widely performed to analyze not only the frequency of immune cells, but also their effector functions as determined by their cytokine production/secretion. The present study aimed to develop a reader-free ELISPOT assay using a handy membrane-punching device termed ELI 8. With the assistance of particle analysis by ImageJ software, the results of spot counting were reproducible with high inter-assay and inter-examiner concordance. Immune cells that produce and secrete Th1 cytokines without antigen-peptide stimulation of peripheral blood mononuclear cells (PBMCs) were detected, and their frequencies in patients with cancer were significantly higher compared with those in healthy individuals. These frequencies varied between individuals, as well as between time points during the course of cancer vaccine immunotherapy in each patient. Due to the variability in spontaneous cytokine production/secretion by PBMCs, an antigen-specific immune response (IR) index is proposed, which is a ratio of the number of spot-forming cells (SFCs) subjected to antigen-stimulation to that of SFCs with spontaneous cytokine secretion without antigen-stimulation. This index may be used as a marker for antigen-specific cellular immune responses in patients treated with cancer immunotherapy. The IR index successfully detected the induction of Wilms' tumor 1-specific cellular immune responses in patients with cancer treated with cancer vaccine immunotherapy.
Identifiants
pubmed: 32257187
doi: 10.3892/br.2020.1289
pii: BR-0-0-1289
pmc: PMC7100143
doi:
Types de publication
Journal Article
Langues
eng
Pagination
244-250Informations de copyright
Copyright © 2020, Spandidos Publications.
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