We found that the number of T cells attached to cancer cells reduces the likelihood that the cancer cell will proliferate, with the beneficial KO T cells having greater effects on proliferation reduction.
Cancer cell and T cell morphology changes dramatically depending on state. These changes are visible in the brightfield imaging – active interacting T cells are larger and change to less circular shapes. Cancer cell begin to aggregate together when interacting with T cells.
While the # of T cell--cancer cell interactions increased similarly, these interactions & their effects were modulated by the CRISPR KOs. E.g., the time a T cell remained attached to a cancer cell (as estimated by a negative binomial and Markov model separately) was highest in RASA2 KO T cells.
Most thrilling is that we can identify active T cells based on relative cell size and morphology, and watch T cells activate (differentially based on condition) after interacting with cancer cells.
With a Markov model, we deconvolved when, in frame t-1, there is one cancer cell and one T cell in a window, and in frame t there is one cancer cell and two T cells. We were able to quantify how often this doubling of T cells attacking a cancer cell was due to proliferation or due to recruitment.
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