Reader‑free ELISPOT assay for immuno‑monitoring in peptide‑based cancer vaccine immunotherapy

  • Authors:
    • Sae Hayashi
    • Rin Imanishi
    • Mayuko Adachi
    • Sayaka Ikejima
    • Jun Nakata
    • Soyoko Morimoto
    • Fumihiro Fujiki
    • Sumiyuki Nishida
    • Akihiro Tsuboi
    • Naoki Hosen
    • Hiroko Nakajima
    • Kana Hasegawa
    • Yoshihiro Oka
    • Haruo Sugiyama
    • Yusuke Oji
  • View Affiliations

  • Published online on: March 5, 2020     https://doi.org/10.3892/br.2020.1289
  • Pages: 244-250
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

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.

Related Articles

Journal Cover

May 2020
Volume 12 Issue 5

Print ISSN: 2049-9434
Online ISSN:2049-9442

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
APA
Hayashi, S., Imanishi, R., Adachi, M., Ikejima, S., Nakata, J., Morimoto, S. ... Oji, Y. (2020). Reader‑free ELISPOT assay for immuno‑monitoring in peptide‑based cancer vaccine immunotherapy. Biomedical Reports, 12, 244-250. https://doi.org/10.3892/br.2020.1289
MLA
Hayashi, S., Imanishi, R., Adachi, M., Ikejima, S., Nakata, J., Morimoto, S., Fujiki, F., Nishida, S., Tsuboi, A., Hosen, N., Nakajima, H., Hasegawa, K., Oka, Y., Sugiyama, H., Oji, Y."Reader‑free ELISPOT assay for immuno‑monitoring in peptide‑based cancer vaccine immunotherapy". Biomedical Reports 12.5 (2020): 244-250.
Chicago
Hayashi, S., Imanishi, R., Adachi, M., Ikejima, S., Nakata, J., Morimoto, S., Fujiki, F., Nishida, S., Tsuboi, A., Hosen, N., Nakajima, H., Hasegawa, K., Oka, Y., Sugiyama, H., Oji, Y."Reader‑free ELISPOT assay for immuno‑monitoring in peptide‑based cancer vaccine immunotherapy". Biomedical Reports 12, no. 5 (2020): 244-250. https://doi.org/10.3892/br.2020.1289