Hi, everyone, i am a fresher of tumor immunology.
Recently, i am preparing to study how pdl1 regulation in tumor cells will affect the function of t cells. and i have read a paper " miR-424(322) reverses chemoresistance via T-cell immune response activation by blocking the PD-L1 immune checkpoint. Nat Commun" . In this paper, author applied PBMC from healthy human with the anti-CD3, anti-CD28 and incubation of skov3 tumor lysates. Then, isolated CD8 T cells to co-culture with skov3 cells. i would like to apply this model, but i have a question, anti-CD3 and anti-CD28 has made CD8+ T cells activated, why do they need to add tumor lysates?
And, right now, i don't make sure whether i should apply antigen-specific CD8+T cells to coculture with tumor cells? other papers indicated that firstly isolated monocyte-derived DC and then make them matured with some cytokine stimulation and tumor lysates incubation. Then matured DC cocultured with CD8+ T cells to get the tumor-antigen specific T cells. These obtained CD8 T cells then cocultured with tumor cell lines. could you kindly give me some advice based on my research purpose?
Hello Jiangnan He
why do they need to add tumor lysates?
We need to add tumor lysates to optimize the ability of the cells to generate tumor-specific T-cell responses. Thus, cells can be loaded either with whole tumor cells or tumor cell lysate, or tumor antigen-enriched fractions to achieve the desired anti-tumor T-cell response. Tumor cell lysate represents the whole protein content of lysed tumor cells. So, the advantage of using tumor lysate is that the multiple antigens heterogeneously expressed on growing tumors can sensitize T-cells.
DCs are the most effective type of antigen-presenting cells and can stimulate naive T lymphocytes to initiate an immune response. DCs represent the interface between the universe of foreign and tissue-specific antigens and T lymphocytes. They also play a role in all aspects of T-cell responses. I would suggest you use monocyte-derived DCs in your study as it will help in a better response.
Best Wishes.
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