Isoplexis_July2021_AcceleratingDevelopmentCurativeMedicines - 10

Identifying Key Drivers of Transplant Rejection with Single-Cell Immune Profiling
suppressed augmentation of CD8+, with a lesser
effect on CD4+ TEM expansion. The researchers
also profiled CD4+ and CD8+ TEM which were not
co-cultured with ECs as controls and used IsoPlexis'
functional single-cell proteomics to analyze the
controls and the treated cells. The control groups
did not produce any cytokines, suggesting that
co-culture with ECs was the factor driving cytokine
production in the proliferated populations.
IsoPlexis' platform identified several distinct
subsets of polyfunctional allogeneic CD4+ and CD8+
TEM
cells. The IsoSpeak software suite was used to
generate advanced visualizations, such as t-SNE,
as well as perform analyses such as calculating the
percentage of polyfunctional cells. The control T
cells, which were stimulated by IFN-γ-primed ECs
showed low levels of polyfunctionality. In contrast,
allogeneic CD4+ and CD8+ TEM
cells were found
to be highly polyfunctional and were significantly
upregulated after simulation by PRA-activated
ECs. While both control and PRA-activated groups
contained non-secretors, the T cells stimulated
by PRA-activated ECs showed higher levels of
polyfunctionality.
Many T cells in the PRA-activated group secreted
>3 cytokines, while the control T cells secreted
less than 3. Additionally, the secretions of the T
cells that were stimulated by PRA-activated ECs
showed stronger signal intensity. Individual T cells
10 | GENengnews.com
Advanced visualizations from the IsoSpeak software
enable stratification of distinct functional cell subsets
with unique cytokine signatures, revealing increased
polyfunctional TEM
by alloimmune responses.
secreted more IFN-γ and TFN-α after stimulation with
PRA-treated ECs compared to controls. Additionally,
CD4+ TEM cultured with PRA-activated ECs secreted
additional cytokines, IL-4 and MIP-1β. Thus, the data
suggest that PRA activation of ECs promotes the
secretion of more cytokines with stronger signal
intensities in CD4+ and CD8+ TEM
cells.
Reducing Drivers of Polyfunctionality and
Decreasing Markers of Transplant Rejection
IsoPlexis' single-cell proteomics revealed that
treating IFN-γ-primed human ECs with PRA significantly
increased the stimulatory effects on CD4+
and CD8+ TEM
proliferation and polyfunctionality,
contributing to higher rates of allograft rejection. The
study also found that treating the ECs with anti-IL-15
antibody reduced this increase in polyfunctionality,
suggesting decreased rates of rejection. The use of
anti-IL-15 blocking antibody significantly reduced
with cellular heterogeneity promoted
http://www.GENengnews.com

Isoplexis_July2021_AcceleratingDevelopmentCurativeMedicines

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Isoplexis_July2021_AcceleratingDevelopmentCurativeMedicines - Contents
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