Ifn-gamma Elispot Assay
Ifn-gamma Elispot Assay - Related Videos
11:53
ELISPOT Assay: Detection of IFN-γ Secreting Splenocytes
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2023
Source: Tonya J. Webb1 1 Department of Microbiology and Immunology, University of Maryland School of Medicine and the Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, Maryland 21201 ELISPOT is a standardized, reproducible assay used to detect cellular immune responses. The assay utilizes an enzyme-linked immunosorbent assay (ELISA)- based method to detect single-cell immune responses which can be visualized by spots, hence the name ELISPOT. ELISPOT was first described in...
08:04
Development of an IFN-γ ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation
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Cited by 9 •
2014
Novel generations of functional assays such as gamma interferon (IFN-γ) ELISpot, which detect cytokine production at the single cell level and provide both quantitative and qualitative characterization of T cell responses can be used to assess cell-mediated immune responses directed against varicella zoster virus (VZV).
04:20
Functional Assessment of Antibody-Secreting B Cells Using ELISpot Assay
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2025
This video demonstrates the use of the ELISpot assay to assess the function of antibody-secreting B cells, ASCs. The antibodies secreted by the ASCs interact with the membrane-bound polyclonal antibody fragments, which are detected by the enzyme-conjugated IgG or IgM antibodies. The colored spot produced in each well indicates the presence of the specific antibody's isotype, which correlates with the B cells' secretory activity.
05:28
Assay for the Identification of Interferon-Gamma Secretion in Murine Lymphoid Organs
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2025
This video demonstrates an imaging method to visualize in situ interferon-gamma (IFN-γ) secretion in the mouse spleen. Following bacterial infection and the induction of IFN-γ accumulation inside T cells and natural killer cells, the spleen is harvested from the mouse. Upon obtaining tissue sections, immunostaining is performed to visualize IFN-γ inside the cells.
08:13
Optimized Interferon-gamma ELISpot Assay to Measure T Cell Responses in the Guinea Pig Model after Vaccination
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Cited by 3 •
2019
The development of the interferon-gamma ELISpot assay for guinea pig PBMCs allows the characterization of T-cell responses in this highly relevant model for studying infectious diseases. We have applied the assay to measure T cell responses associated with intradermal delivery of DNA vaccines.
FAQs
Capture antibodies immobilized on the membrane retain interferon-gamma close to the secreting cell instead of allowing the cytokine to disperse throughout the culture. Labeled detection antibodies then identify the captured cytokine, and visualization produces localized spots. This arrangement allows the assay to reveal responses from individual cells and supports sensitive measurement of cellular immune activity.
Each visible spot corresponds to an individual immune cell that released interferon-gamma under the assay conditions. Consequently, spot counts provide an estimate of the frequency of responding cells rather than only a pooled cytokine concentration. Comparing responses to antigen stimulation can therefore indicate how extensively a sample contains cells capable of mounting that cellular immune response.
Antigen stimulation provides the defined immune trigger used to assess whether cells respond to a particular antigen. The resulting interferon-gamma release reflects antigen-responsive cellular activity, allowing investigators to examine functional immunity rather than simply detect the presence of immune cells. This distinction is especially relevant when studying responses to infectious agents or evaluating vaccine-induced cellular immunity.
The assay links cytokine release to the frequency of individual responding cells. Its readout therefore helps characterize how many cells participate in an antigen-specific response and whether those cells display measurable interferon-gamma-producing function. This cell-level perspective complements broader assessments of cellular immunity and helps researchers interpret differences in immune responsiveness between experimental samples.
The workflow begins by placing antigen-stimulated immune cells onto a membrane coated with capture antibodies. Secreted interferon-gamma is captured near the cells, followed by detection with labeled antibodies. A visualization step produces spots for the captured cytokine. Researchers then use the resulting spot pattern and count to assess the frequency and functional activity of responding cells.
The essential components described for the method are antigen-stimulated immune cells, a membrane coated with interferon-gamma capture antibodies, labeled detection antibodies, and a visualization system that reveals the spots. Each component contributes a different function: cells provide the response, antibodies capture and identify the cytokine, and visualization converts secretion events into an observable assay readout.
Researchers apply the assay when they need to measure antigen-specific cellular immune activity in contexts such as infection, vaccination, and immune monitoring. Because the readout reflects both the frequency and function of cytokine-producing cells, it can support studies of how hosts respond to antigens and how cellular immunity changes across experimental or clinical investigations.
In infection-focused immunology, antigen stimulation can be used to examine whether immune cells produce interferon-gamma in response to pathogen-associated antigens. The resulting spot frequency provides a cellular measure of antigen-responsive activity, helping investigators assess host cellular immunity. These data can contribute to understanding immune responses during host-pathogen interactions and to evaluating infection-related immune monitoring outcomes.