Abstract
ABSTRACTCytoskeletal actin dynamics are crucial for the activation of T-cells. Immortalised Jurkat T-cells have been the model system of choice to examine and correlate the dynamics of the actin cytoskeleton and the immunological synapse leading to T-cell activation. However, it has remained unclear whether immortalised cellular systems, such as Jurkat T-cells can recapitulate the cytoskeletal behaviour of primary T-cells. Studies delineating the cytoskeletal behaviour of Jurkat T-cells in comparison to primary T-cells are lacking. Here, we employ live-cell super-resolution microscopy to investigate the cytoskeletal actin organisation and dynamics of living primary and immortalised Jurkat T-cells at the appropriate spatiotemporal resolution. Under comparable activation conditions, we found differences in the architectural organisation and dynamics of Jurkat and primary mouse and human T-cells. Although the three main actin network architectures in Jurkat T-cells were reminiscent of primary T-cells, there were differences in the organisation and molecular mechanisms underlying these networks. Our results highlight mechanistic distinctions in the T-cell model system most utilised to study cytoskeletal actin dynamics.
Highlights
A healthy actin cytoskeleton is crucial for T-cell function
Quantitative comparison of calcium (Ca2+) triggering of large T-cell ensembles of all three cellular systems did not indicate significant statistical differences in the Ca2+-triggering fractions but a slowdown in the Ca2+ response time of Jurkat CD4+ T-cells compared to primary CD4+ T-cells (Fig. S1)
The lamellar leading edge was more dynamic in mouse primary T-cells, as reflected by significantly higher mean curvature magnitude and persistent fluctuations compared to those in the Jurkat T-cells (Fig. 1E)
Summary
A healthy actin cytoskeleton is crucial for T-cell function. The actin machinery integrates T-cell receptor (TCR) signalling and biophysical mechanisms to coordinate the activation of T-cells at the immunological synapse (IS), for T-cell activation, function and differentiation (Colin-York et al, 2019a; Fritzsche et al, 2017; Roy and Burkhardt, 2018). We found differences in the actin organisation and dynamics between Jurkat cells, an extensively utilised immortalised T-cell system, and primary mouse and human T-cells at comparable activation conditions. Quantitative comparison of calcium (Ca2+) triggering of large T-cell ensembles of all three cellular systems did not indicate significant statistical differences in the Ca2+-triggering fractions but a slowdown in the Ca2+ response time of Jurkat CD4+ T-cells compared to primary CD4+ T-cells (Fig. S1).
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