ArticleJournal of immunological methods2012
Optimization of a whole blood intracellular cytokine assay for measuring innate cell responses to mycobacteria.
Article in Journal of immunological methods, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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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
8 citing papers in PubMed, 13 citations in OpenAlex.
- Maturation of innate responses to mycobacteria over the first nine months of life.Journal of immunology (Baltimore, Md. : 1950) · 2014Trial
- Measuring single-cell protein secretion in immunology: Technologies, advances, and applications.European journal of immunology · 2021Review
- High-throughput analysis of T cell-monocyte interaction in human tuberculosis.Clinical and experimental immunology · 2020Article
- Cell-Mediated Immune Responses toFrontiers in immunology · 2020Article
- In vitro analysis of antigen induced T cell-monocyte conjugates by imaging flow cytometry.Journal of immunological methods · 2018Article
- Immune cell dysfunctions in breast cancer patients detected through whole blood multi-parametric flow cytometry assay.Oncoimmunology · 2016Article
- Measuring Cellular Immunity to Influenza: Methods of Detection, Applications and Challenges.Vaccines · 2015Review
- Ontogeny of Toll-like receptor mediated cytokine responses of South African infants throughout the first year of life.PloS one · 2012Article
Corrections and comments
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Authors and funding
8 authors at 3 institutions in 2 countries.
Funding
Abstract
Innate cells are essential for host defense against invading pathogens, and the induction and direction of adaptive immune responses to infection. We developed and optimized a flow cytometric assay that allows measurement of intracellular cytokine expression by monocytes, dendritic cells (DC) and granulocytes, as well as cellular uptake of green-fluorescent protein (GFP)-expressing mycobacteria, in very small volumes of peripheral blood. We show that innate cell stimulation resulted in increased granularity of monocytes and mDC and decreased granulocyte granularity that precluded flow cytometric discernment of granulocytes from monocytes and myeloid DC by forward and side scatter gating. Anti-CD66a/c/e antibody staining allowed reliable identification and exclusion of granulocytes for subsequent delineation of monocytes and myeloid DC. Intracellular cytokine expression by granulocytes, monocytes and mDC was remarkably sensitive to the dose of mycobacterial inoculum. Moreover, activation of monocytes and mDC with live BCG reduced expression levels of CD14 and CD11c, respectively, necessitating optimization of staining conditions to reliably measure these lineage markers. Finally, we characterized expression of IL-12/23p40, TNF-α, IL-6, and IL-10, by GFP(+) and GFP(-) monocytes and mDC from 25 healthy adults. This assay may be applied to the study of innate cell responses to any GFP-expressing pathogen, and can be performed on blood volumes as low as 200 μL per condition, making the assay particularly suitable for pediatric studies.
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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.