PMID- 34053051
OWN - NLM
STAT- MEDLINE
DCOM- 20210816
LR  - 20210816
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2325
DP  - 2021
TI  - CTL ELISPOT Assay and T Cell Detection.
PG  - 65-77
LID - 10.1007/978-1-0716-1507-2_5 [doi]
AB  - Enzyme-linked immune absorbent spot (Elispot) is a quantitative method for 
      measuring relevant parameters of T-cell activation. The sensitivity of Elispot 
      allows the detection of low-frequency antigen-specific T-cells that secrete 
      cytokines and effector molecules, such as granzyme B and perforin. Cytotoxic 
      T-cell (CTL) studies have taken advantage with this high-throughput technology by 
      providing insights of quantity and immune kinetics. Accuracy, sensitivity, 
      reproducibility, and robustness of Elispot resulted in a wide range of 
      applications in research as well as in diagnostic field. Actually, CTL monitoring 
      by Elispot is a gold standard for the evaluation of antigen-specific T-cell 
      immunity in clinical trials and vaccine candidates where the ability to detect 
      rare antigen-specific T-cells is of relevance for immune diagnostic. The most 
      utilized Elispot assay is the Interferon-gamma (IFN-gamma) test, a marker for CD8(+) 
      CTL activation, but Elispot can be also used to distinguish different subsets of 
      activated T-cells by using other cytokines such as T-helper (Th) 1 type cells 
      (characterized by the production of IFN-gamma, IL-2, IL-6, IL-12, IL-21 and TNF-alpha), 
      Th2 (producing cytokines like IL-4, IL-5, IL-10 and IL-13), and Th17 (IL-17) 
      cells.The reliability of Elispot generated data, by the evaluation of T-cell 
      frequency recognizing individual antigen/peptide, is the core of this method 
      currently applied widely to investigate specific immune responses in cancer, 
      infections, allergies, and autoimmune diseases. The Elispot Assay is competing 
      with other methods measuring single-cell cytokine production, e.g., intracellular 
      cytokine by FACS or Milteny cytokine secretion assay. Other types of lymphocyte 
      frequency and function assays include limiting dilution assay (LDA), cytotoxic 
      T-cell assay (CTL), and tetramer staining. Compared with respect to sensitivity 
      the Elispot Assay is outranking other methods to define frequency of 
      antigen-specific lymphocytes. The method described herein would like to offer 
      helpful and clear protocols for researchers that apply Elispot. IFN-gamma and 
      Perforin Elispot assays will be described.
FAU - Ranieri, Elena
AU  - Ranieri E
AD  - Center of Molecular Medicine, Clinical Pathology, Department of Surgical and 
      Medical Sciences, University of Foggia, Foggia, Italy. elena.ranieri@unifg.it.
FAU - Netti, Giuseppe Stefano
AU  - Netti GS
AD  - Center of Molecular Medicine, Clinical Pathology, Department of Surgical and 
      Medical Sciences, University of Foggia, Foggia, Italy.
FAU - Gigante, Margherita
AU  - Gigante M
AD  - Center of Molecular Medicine, Clinical Pathology, Department of Surgical and 
      Medical Sciences, University of Foggia, Foggia, Italy.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Cytokines)
RN  - 126465-35-8 (Perforin)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - CD8-Positive T-Lymphocytes/*metabolism
MH  - Cytokines/*metabolism
MH  - Enzyme-Linked Immunospot Assay/*methods
MH  - Humans
MH  - Interferon-gamma/metabolism
MH  - Perforin/metabolism
OTO - NOTNLM
OT  - Cancer
OT  - Cytokines
OT  - ELISPOT
OT  - T-cell
EDAT- 2021/05/31 06:00
MHDA- 2021/08/17 06:00
CRDT- 2021/05/30 21:12
PHST- 2021/05/30 21:12 [entrez]
PHST- 2021/05/31 06:00 [pubmed]
PHST- 2021/08/17 06:00 [medline]
AID - 10.1007/978-1-0716-1507-2_5 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2021;2325:65-77. doi: 10.1007/978-1-0716-1507-2_5.