ArticleCytometry. Part A : the journal of the International Society for Analytical Cytology2010
Ki-67 staining for determination of rhesus macaque T cell proliferative responses ex vivo.
Article in Cytometry. Part A : the journal of the International Society for Analytical Cytology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Syndecan-1 Regulates Psoriasiform Dermatitis by Controlling Homeostasis of IL-17-Producing γδ T Cells.Journal of immunology (Baltimore, Md. : 1950) · 2018Article
- In vivo inhibition of tryptophan catabolism reorganizes the tuberculoma and augments immune-mediated control ofProceedings of the National Academy of Sciences of the United States of America · 2018Article
- Beyond Passive Immunity: Is There Priming of the Fetal Immune System Following Vaccination in Pregnancy and What Are the Potential Clinical Implications?Frontiers in immunology · 2018Review
- Adoptive Transfer of Phosphoantigen-Specific γδ T Cell Subset AttenuatesJournal of immunology (Baltimore, Md. : 1950) · 2017Article
- A Single Dose Respiratory Recombinant Adenovirus-Based Vaccine Provides Long-Term Protection for Non-Human Primates from Lethal Ebola Infection.Molecular pharmaceutics · 2015Article
- Pathophysiology of hantavirus pulmonary syndrome in rhesus macaques.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- Re-evaluation of PD-1 expression by T cells as a marker for immune exhaustion during SIV infection.PloS one · 2013Article
- Pushing the frontiers of T-cell vaccines: accurate measurement of human T-cell responses.Expert review of vaccines · 2012Review
- Synthetic DNA immunogen encoding hepatitis B core antigen drives immune response in liver.Cancer gene therapy · 2012Article
- HIV-mediated phosphatidylinositol 3-kinase/serine-threonine kinase activation in APCs leads to programmed death-1 ligand upregulation and suppression of HIV-specific CD8 T cells.Journal of immunology (Baltimore, Md. : 1950) · 2011Article
- Article
- Novel application of Ki67 to quantify antigen-specific in vitro lymphoproliferation.Journal of immunological methods · 2010Article
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Authors and funding
7 authors.
Funding
Abstract
The capacity for robust proliferation upon re-infection is a hallmark of adaptive immunity and the basis of vaccination. A widely used animal model for the study of human disease is the rhesus macaque (RM), where capacity for proliferation can be assessed ex vivo using carboxyfluorescein succinimidyl ester (CFSE)-based dilution assays. However, we show over the course of the standard ex vivo proliferation assay that CFSE-labeling at commonly used dye concentrations induces significant cell death, but that this phenomenon is dose-dependent. Here, we describe an alternative semiquantitative method for estimating T cell proliferative responses that avoids the putative biases associated with chemical modification. RM peripheral blood mononuclear cells were stimulated ex vivo with cognate peptides for 5 days, immunostained for intracellular Ki-67, and then analyzed by flow cytometry. We describe a gating strategy using Ki-67 and side light scatter, also a marker of blastogenesis, which correlates strongly with data from CFSE dilution. We show that this method is a valid tool for measuring RM antigen-specific cellular proliferation ex vivo and can be used as an alternative to CFSE dilution assays.
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