Immuno-Oncology Advances in Applied Cellular Analysis - 21

Advances in Applied Cellular Analysis * Continuous Live-Cell Analysis of the Immune-Tumor Axis

Trastuzumab (Herceptin®) is a clinically used
mAb treatment for HER2-positive solid cancers.
To compare the antibody-dependent (tumor)
cell killing (ADCC) of trastuzumab and potential
biosimilar mAbs, IncuCyte CLCA assays were
assembled with co-cultures of HER2-positive
SKOV-3 ovarian cancer cells and peripheral blood
mono-nuclear cells (Figures 4, 5). SKOV-3 cells
were stably transduced with IncuCyte NucLight
Red and plated at 5K cells per well on 96-well
plates. PBMCs (25K cells per well) were added at
t=0h. Apoptotic tumor cells were enumerated
using IncuCyte Caspase 3/7 green substrate,
a mix and read no-wash reagent suitable for
live cell imaging.2 IncuCyte images were taken
every 2h and automatically analyzed for red
(tumor count) and green (apoptotic cell count)
objects. In control (vehicle-treated) co-cultures,
SKOV-3 cells proliferated over time (0-140h) and
there was little or no observable apoptosis until
post 100h. Activating PBMCs with anti-CD3/IL-2
caused cell killing as measured by a reduction in
both tumor cell count and increase in apoptotic
nuclei. Trastuzumab also produced a timeand concentration-dependent inhibition of
21

| GENengnews.com

Figure 4. Additional biological insight with IncuCyte continuous live-cell analysis.
Sequential image montage of immune-cell killing (PBMCs) of SKOV-3 ovarian cancer cells. Engagement of cancer cell by
immune cells (1) and (2), tumor cell death, granulation & green caspase-3/7 signal (3), and tumor cell division (4).

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Immuno-Oncology Advances in Applied Cellular Analysis

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