Human T Cell Quantification
Human T Cell Quantification - Related Videos
10:18
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Cited by 7 •
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Dendritic spines of pyramidal neurons are the sites of most excitatory synapses in mammalian brain cortex. This method describes a 3D quantitative analysis of spine morphologies in human cortical pyramidal glutamatergic neurons derived from induced pluripotent stem cells.
03:45
Quantification of Inflammatory Mediators in Infected Human Tissue-Derived Cells
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This video demonstrates the quantification of inflammatory mediators during infection by incubating human lung-tissue-derived immune cells with Haemophilus influenzae. Flow cytometry is used to measure cytokine fluorescence in labeled lymphocytes, correlating with the production of pro-inflammatory cytokines in response to bacterial infection.
08:26
The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity
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Cited by 10 •
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Human peripheral blood is commonly used for the assessment of the humoral immune response. Here, the methods for isolating human B cells from peripheral blood, differentiating human B cells into antibody (Ab)-secreting B cells (ASCs) in culture, and enumerating the total IgM- and IgG-ASCs via an ELISpot assay are described.
14:03
Transduction of Human Cells with Polymer-complexed Ecotropic Lentivirus for Enhanced Biosafety
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Cited by 8 •
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Lentiviruses are a valuable research tool for exploring gene function; however, researchers may wish to avoid production of pantropic lentivirus encoding known or suspected oncogenes. As an alternative, we present a safer protocol for use of ecotropic lentivirus on human cells modified to express the ecotropic receptor mSlc7a1.
07:53
Quantification of Antibody-dependent Enhancement of the Zika Virus in Primary Human Cells
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Cited by 1 •
2019
We describe a method to evaluate the effect of pre-existing immunity against dengue virus on the Zika virus infection by using human serum, primary human cells, and infection quantification by quantitative real-time polymerase chain reaction.
FAQs
Sequential gating narrows the analyzed population in stages, allowing fluorescently labeled cells to be identified according to the markers they carry. CD3 helps identify T lymphocytes, while CD4 and CD8 separate major T-cell subsets within that population. This organization reduces ambiguity when interpreting total, helper, and cytotoxic compartments across biological samples.
Using the markers together provides layered classification rather than a single total. CD3 identifies the T-lymphocyte population, while CD4 or CD8 reveals its major subset composition. A sample can therefore show stable overall T-cell abundance while displaying a changed balance between helper and cytotoxic populations, an outcome relevant to immune responses during infection.
Frequency reports the proportion of analyzed cells assigned to a T-cell population, whereas an absolute count expresses the number of cells in the measured sample. Counting beads or a measured sample volume enables this conversion. Absolute values can therefore reveal changes in cellular abundance that a percentage alone might not show.
An analysis typically applies fluorescent antibodies to the sample, then uses flow cytometry to detect labeled cells. Sequential gating organizes the measurements so that T lymphocytes and their CD4- or CD8-defined subsets can be counted. Counting beads or measured sample volume can then support conversion from cellular frequency to an absolute number.
Measurements can characterize immune dysfunction, compare cellular responses during infection, and follow changes associated with treatment or vaccination. They can also support assessment of disease progression and recovery. Because the assay distinguishes total T cells from CD4- and CD8-defined populations, investigators can examine whether immune changes affect broad abundance or particular subsets.
Comparing CD4 and CD8 results helps determine whether an observed change is distributed across major T-cell subsets or concentrated in one population. This distinction adds context to total T-cell counts: two samples may have similar overall abundance but different helper and cytotoxic composition. In infection studies, that comparison supports more precise characterization of cellular immune responses.