With the masked, tracked cells, we went to work to develop Occident. We were curious how well the RFP markers captured cancer cell number; we found that RFP lags as a proxy for cancer cell numbers.
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.
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