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Detecting interferon-gamma release from human CD4 T-cells using surface plasmon resonance
Gulnaz Stybayeva1, Markhabat Kairova, Erlan Ramanculov
1Department of Biomedical Engineering, University of California, Davis, CA 95616, United States.
Insights
This study presents a novel method combining cell separation and surface plasmon resonance (SPR) to detect cytokine production. The technique accurately links interferon-gamma secretion to specific CD4 T-cells, aiding disease diagnostics.
Area of Science:
- Immunology
- Biotechnology
- Biochemistry
Background:
- Leukocyte-derived cytokines are crucial indicators of immune responses and disease diagnostics.
- Identifying specific leukocyte subsets responsible for cytokine secretion is challenging due to cellular heterogeneity.
- Accurate attribution of cytokine production to distinct immune cell populations is vital for understanding immune function and disease pathogenesis.
Purpose of the Study:
- To develop and validate a combined strategy for isolating specific leukocyte subsets and detecting their secreted cytokines.
- To enable label-free quantification of interferon-gamma (IFN-γ) secreted by CD4 T-cells.
- To establish a foundation for sensitive, subset-specific cytokine detection in biological samples.
Main Methods:
- Antibody (Ab)-based affinity cell separation was employed to capture human CD4 T-cells within a flow chamber.
- Cells were activated in situ, and the secreted cytokines in the culture medium were analyzed.
- Surface Plasmon Resonance (SPR) with immobilized anti-IFN-γ monoclonal antibodies was used for label-free detection of secreted IFN-γ.
Main Results:
- The developed method successfully captured CD4 T-cells from human blood.
- SPR signal intensity directly correlated with the amount of IFN-γ produced by the activated CD4 T-cells.
- This demonstrates the feasibility of linking specific cytokine secretion to purified cell populations.
Conclusions:
- The combined approach of SPR detection and cell purification offers a sensitive, label-free method for analyzing cytokine production.
- This strategy can be extended for the detection of various cytokines or proteins secreted by specific immune cell subsets.
- This technique holds promise for advancing disease diagnostics and immunological research by providing precise information on immune cell function.
Abstract:
Cytokine secretion by leukocytes is an important indicator of immune response to pathogens and therefore has significant implications in disease diagnostics. Given heterogeneity of leukocyte subsets and the ability of multiple cell subsets to secrete the same cytokines, connecting cytokine production to a specific leukocyte subset is a distinct challenge. In the present paper we describe a strategy combining antibody (Ab)-based affinity cell separation and surface plasmon resonance (SPR) for capturing human CD4 T-cells and for label-free detection of cell-secreted interferon (IFN)-gamma--an important inflammatory cytokine. Human blood was introduced into a flow chamber modified with anti-CD4 Abs resulting in capture of CD4(+) T-cells. After mitogenic activation of cells inside the flow chamber, culture medium was routed onto an SPR chip modified with monoclonal IFN-gamma Abs. SPR signal observed in this experiment correlated with cytokine production by T-cells. The strategy of combining SPR detection with cell purification may be used in the future for label-free, sensitive detection of multiple cytokines or proteins secreted by the desired cell subset.
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