Supplementary Materials

Supramolecular attack particles are autonomous killing entities released from cytotoxic T cells

Š. Balint, S. Müller, R. Fischer, B. M. Kessler, M. Harkiolaki, S. Valitutti, M. L. Dustin

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S20
  • Captions for Movies S1 to S14
  • Caption for Data S1
  • References
Data S1
List of proteins present in CD8+ T-cell released SMAPs identified by mass spectrometry. Unfiltered list of proteins present in CD8+ T-cell released SMAPs under activating (ICAM-1 + anti-CD3ε light blue) or non-activating (ICAM-1; light green) conditions, identified by mass spectrometry (n=3 donors).
MDAR Reproducibility Checklist

Images, Video, and Other Media

Movie S1
Live confocal imaging of the transfer of Gzmb-mCherry+ (green) and WGA (magenta) labeled SMAPs from an antigen specific CTL clone into pp65-pulsed JY target cells. BF, bright field microscopy. Frame rate, 3 frames/min.
Movie S2
3D confocal z-stack projection depicting the transfer of Gzmb-mCherry+ (green) and WGA (magenta) labeled SMAPs from an antigen-specific CTL clone into pp65-pulsed JY target cells (cyan). BF, bright field microscopy.
Movie S3
3D confocal z-stack projection depicting the absence of the transfer of Gzmb-mCherry+ (green) and WGA (magenta) labeled SMAPs from an antigen-specific CTL clone into unpulsed JY target cells (cyan). BF, bright field microscopy.
Movie S4
Live TIRFM imaging of the release of SMAPs from Gzmb-mCherry-SEpHluorin (magenta/green) transfected CD8+ T-cells on activating (ICAM-1 (blue) + anti-CD3ε) SLB. Frame rate, 2 frames/min.
Movie S5
Schematic of the working model for capturing SMAPs released by activated CD8+ T-cells. CD8+ T-cells (grey) were incubated on activating SLB for the desired time. After incubation, cells were removed with cold PBS leaving the released SMAPs (purple) on the SLB. Elements are not drawn to a scale.
Movie S6
Live TIRFM imaging of the release of Prf1+ (green) SMAPs by CD8+ T-cells after contact with activating (ICAM-1 (blue) + anti-CD3ε) SLB. Frame rate, 1 frame/min.
Movie S7
Live TIRFM imaging of the release of Gzmb+ (red) SMAPs by CD8+ T-cells after contact with activating (ICAM-1 (blue) + anti-CD3ε) SLB. Frame rate, 1 frame/min.
Movie S8
Live TIRFM imaging of the release of SMAPs double positive for Prf1 (green) and Gzmb (magenta) by CD8+ T-cells after contact with activating (ICAM-1 (blue) + anti-CD3ε) SLB. Frame rate, 1 frame/min.
Movie S9
Live TIRFM imaging of the release of SMAPs double positive for Prf1 (green) and Gzmb (magenta) by CD8+ T-cells after contact with activating (ICAM-1 (blue) + anti-CD3ε) SLB. Frame rate, 1 frame/min.
Movie S10
Live TIRFM imaging of the release of Prf1+ (green) and TSP-1+ (magenta) SMAPs by CD8+ T-cells after contact with activating (ICAM-1 (blue) + anti-CD3ε) SLB. Frame rate, 1 frame/min.
Movie S11
3D confocal z-stack projection and orthogonal views of CD8+ T cells co-transfected with Gzmb-mCherry-SEpHluorin (magenta) and TSP-1-GFPSpark (green) on non-activating (ICAM-1; left) or activating (ICAM-1 + anti-CD3ε; right) SLB. Cells were stained with WGA (yellow) to visualize the cell membrane. The formation of a mature IS is indicated by an ICAM-1 ring (blue).
Movie S12
CSXT projection and 3D reconstruction of individual released SMAPs captured on a carbon coated EM grid containing ICAM-1 and anti-CD3ε.
Movie S13
CSXT projection and 3D reconstruction of a CD8+ T-cell interacting with a carbon coated EM grid containing ICAM-1 and anti-CD3ε. SMAPs can be observed within the multivesicular bodies inside the cell.
Movie S14
CSXT projection and 3D reconstruction of a CD8+ T-cell interacting with a carbon coated EM grid containing ICAM-1 and anti-CD3ε. Released SMAPs (yellow) can be observed beneath the cell. Nucleus is highlighted in blue.