We have a hypothesis that one of our cancer treatment method can induce specific antitumor immune response. I would like to measure it quantitatively in a simple Colon26 cell line derived allograft tumor model in mouse. Elispot assays are widely used for this purpose, but I am absolutely open for any other method, suggestion or notes.
Thanks in advance.
You can make a co-culture of the local immune cells (from draining lymph nodes) or systemic immune cells (from blood or spleen) with the tumor cell line and measure the cytokines (IFN-g, TNF) from the supernatant by ELISA.
Try for tumor-specific antibodies. Treat tumor-bearing mice, collect sera and perform a cytotoxicity assay (ADCC - so effectors with CD16/32). Also ctx-assay may be performed with PBMC or SMC effectors from treated mice, with Colon26 as target cells for specific response, and other cell lines for non-specific response. You can also try non-treated mice derived effectors to estabilish a baseline. CTX is quantitative and can be performed by either 51Cr assay, or by flow cytometry methods.
you can co-culture treated mice splenocytes with Colon26 cells previously marked with CFSE and check cells viability using a flow cytometer. Is a non-radioactive CTL assay.
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