ArticleJournal of immunological methods2007
A dual color ELISPOT method for the simultaneous detection of IL-2 and IFN-gamma HIV-specific immune responses.
Article in Journal of immunological methods, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
21 citing papers in PubMed, 46 citations in OpenAlex.
- Trial
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- Rational design of multi-epitope vaccine for Chandipura virus using an immunoinformatics approach.PloS one · 2025Article
- Cellular Immunity in Mice Vaccinated with Recombinant Phospholipase D Toxoid ofJournal of arthropod-borne diseases · 2022Article
- A high-throughput screening RT-qPCR assay for quantifying surrogate markers of immunity from PBMCs.Frontiers in immunology · 2022Article
- Non-Assembled ORF2 Capsid Protein of Porcine Circovirus 2b Does Not Confer Protective Immunity.Pathogens (Basel, Switzerland) · 2021Article
- Measuring single-cell protein secretion in immunology: Technologies, advances, and applications.European journal of immunology · 2021Review
- Interferon gamma (IFN-γ) negative CD4+ and CD8+ T-cells can produce immune mediators in response to viral antigens.Vaccine · 2019Article
- CD8+ T-cell mediated anti-malaria protection induced by malaria vaccines; assessment of hepatic CD8+ T cells by SCBC assay.Human vaccines & immunotherapeutics · 2017Article
- Triple Cytokine FluoroSpot Analysis of Human Antigen-Specific IFN-γ, IL-17A and IL-22 Responses.Cells · 2014Article
- Article
- Development of an IFN-γ ELISpot assay to assess varicella-zoster virus-specific cell-mediated immunity following umbilical cord blood transplantation.Journal of visualized experiments : JoVE · 2014Article
- Imaging of Protein Secretion from a Single Cell Using Plasmonic Substrates.BioNanoScience · 2013Article
- Enumeration and characterization of human memory T cells by enzyme-linked immunospot assays.Clinical & developmental immunology · 2013Review
- Cellular barcodes for efficiently profiling single-cell secretory responses by microengraving.Analytical chemistry · 2012Article
- Simultaneous Detection of Antigen-Specific IgG- and IgM-Secreting Cells with a B Cell Fluorospot Assay.Cells · 2012Article
- Article
- ELISpot for measuring human immune responses to vaccines.Expert review of vaccines · 2011Review
- Changes in function of HIV-specific T-cell responses with increasing time from infection.Viral immunology · 2010Article
- The role of IFN-gamma Elispot assay in HIV vaccine research.Nature protocols · 2009Article
Corrections and comments
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Authors and funding
9 authors at 4 institutions in 1 country.
Funding
Abstract
The single color IFN-gamma ELISPOT assay has become a standard for assessing HIV-specific immune responses in HIV-infected subjects. However, recent data suggests that single cytokine detection for immune monitoring of HIV-infected individuals may not be sufficient to fully describe virus-specific immune responses. Here, we have designed and validated a dual color ELISPOT assay capable of detecting both IL-2 and IFN-gamma secreting cells simultaneously in response to HIV antigens. We found that a cell input number of 200,000 cells/well provided a good balance between limited availability of cells due to blood volume restrictions and ability to detect all cytokine secretion patterns. The simultaneous detection of IL-2 and IFN-gamma resulted in a decreased magnitude of IFN-gamma but not IL-2 responses. Measures of intra- and inter-assay variability for the dual color ELISPOT assay were comparable to that seen for single cytokine ELISPOT assay with coefficients of variation below 20% for IL-2, IFN-gamma and dual secretion. Although CD8+ T cells mediated most HIV-specific responses in infected subjects, CD4+ T cells mediated responses to HIV were also detected. Features of this assay such as high throughput, cell number requirement and cytokine choice should make this assay a valuable tool for screening for HIV-specific immune responses in several clinically relevant settings.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.