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High-Content Quantification of Single-Cell Immune Dynamics.

Junkin M, Kaestli AJ, Cheng Z, Jordi C, Albayrak C, Hoffmann A, Tay S - Cell Rep (2016)

Bottom Line: Characterizing dynamic input-output relationships in single cells is crucial for understanding and modeling cellular systems.We developed an automated microfluidic system that delivers precisely defined dynamical inputs to individual living cells and simultaneously measures key immune parameters dynamically.Our system combines nanoliter immunoassays, microfluidic input generation, and time-lapse microscopy, enabling study of previously untestable aspects of immunity by measuring time-dependent cytokine secretion and transcription factor activity from single cells stimulated with dynamic inflammatory inputs.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.

No MeSH data available.


Heterogeneity in Single-Cell Immune Dynamics(A and B) NF-κB (RelA) (A) and TNF secretion (B) in response to 2 hr of LPS (500 ng ml−1). The graphs depict whole population and subpopulation responses from top to bottom, respectively.(C) Total secretion over the course of the entire experiment (14 hr). The line is log-normal fit to data. Data in (B) and (C) are combined from two repeat experiments.
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fig5: Heterogeneity in Single-Cell Immune Dynamics(A and B) NF-κB (RelA) (A) and TNF secretion (B) in response to 2 hr of LPS (500 ng ml−1). The graphs depict whole population and subpopulation responses from top to bottom, respectively.(C) Total secretion over the course of the entire experiment (14 hr). The line is log-normal fit to data. Data in (B) and (C) are combined from two repeat experiments.

Mentions: Single-cell responses (NF-κB translocation, TNF secretion, migration, cell morphology) in all LPS exposure experiments were heterogeneous, and certain subsets of reaction phenotypes could be identified. Figure 5A shows the NF-κB (RelA) response of cells exposed to a 2-hr pulse of LPS. At least four distinct sub-populations were observed in this scenario: those possessing a single NF-κB peak, a single peak followed by a later rise, a single peak and subsequent oscillations, and those that did not respond or only minimally responded to the LPS. TNF secretion could likewise be grouped into subsets with different temporal characteristics in response to LPS (Figure 5B). These consisted of cells that released a single, temporally limited pulse of TNF (within 2 hr), those that released TNF for an extended period (within 4 hr), those that released TNF with other types of profiles such as multi-peak secretion, and those cells that did not release TNF in response to LPS. These subsets were similarly observed when LPS was preceded by 2 hr of IL-10 (20 ng ml−1) (Millipore, IL020) (Figure S7), and similar secretion subsets are observed with other LPS input dynamics. The amount of total TNF produced over 14 hr in response to a 2-hr LPS input is shown in Figure 5C. This followed a skewed distribution with a mean of 4.9 × 106 TNF molecules released, but several cells secreted much higher amounts. These results are typical of the types of dynamic responses exhibited by macrophages and taken in the context of the range of temporal inputs supplied to macrophages shows the dynamic signal processing functionalities of these single cells.


High-Content Quantification of Single-Cell Immune Dynamics.

Junkin M, Kaestli AJ, Cheng Z, Jordi C, Albayrak C, Hoffmann A, Tay S - Cell Rep (2016)

Heterogeneity in Single-Cell Immune Dynamics(A and B) NF-κB (RelA) (A) and TNF secretion (B) in response to 2 hr of LPS (500 ng ml−1). The graphs depict whole population and subpopulation responses from top to bottom, respectively.(C) Total secretion over the course of the entire experiment (14 hr). The line is log-normal fit to data. Data in (B) and (C) are combined from two repeat experiments.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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fig5: Heterogeneity in Single-Cell Immune Dynamics(A and B) NF-κB (RelA) (A) and TNF secretion (B) in response to 2 hr of LPS (500 ng ml−1). The graphs depict whole population and subpopulation responses from top to bottom, respectively.(C) Total secretion over the course of the entire experiment (14 hr). The line is log-normal fit to data. Data in (B) and (C) are combined from two repeat experiments.
Mentions: Single-cell responses (NF-κB translocation, TNF secretion, migration, cell morphology) in all LPS exposure experiments were heterogeneous, and certain subsets of reaction phenotypes could be identified. Figure 5A shows the NF-κB (RelA) response of cells exposed to a 2-hr pulse of LPS. At least four distinct sub-populations were observed in this scenario: those possessing a single NF-κB peak, a single peak followed by a later rise, a single peak and subsequent oscillations, and those that did not respond or only minimally responded to the LPS. TNF secretion could likewise be grouped into subsets with different temporal characteristics in response to LPS (Figure 5B). These consisted of cells that released a single, temporally limited pulse of TNF (within 2 hr), those that released TNF for an extended period (within 4 hr), those that released TNF with other types of profiles such as multi-peak secretion, and those cells that did not release TNF in response to LPS. These subsets were similarly observed when LPS was preceded by 2 hr of IL-10 (20 ng ml−1) (Millipore, IL020) (Figure S7), and similar secretion subsets are observed with other LPS input dynamics. The amount of total TNF produced over 14 hr in response to a 2-hr LPS input is shown in Figure 5C. This followed a skewed distribution with a mean of 4.9 × 106 TNF molecules released, but several cells secreted much higher amounts. These results are typical of the types of dynamic responses exhibited by macrophages and taken in the context of the range of temporal inputs supplied to macrophages shows the dynamic signal processing functionalities of these single cells.

Bottom Line: Characterizing dynamic input-output relationships in single cells is crucial for understanding and modeling cellular systems.We developed an automated microfluidic system that delivers precisely defined dynamical inputs to individual living cells and simultaneously measures key immune parameters dynamically.Our system combines nanoliter immunoassays, microfluidic input generation, and time-lapse microscopy, enabling study of previously untestable aspects of immunity by measuring time-dependent cytokine secretion and transcription factor activity from single cells stimulated with dynamic inflammatory inputs.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosystems Science and Engineering, ETH Zürich, 4058 Basel, Switzerland.

No MeSH data available.