Is there any standard protocol for isolating of tumor-reactive CD8+ T cells from a patient?
I want to have a tumor antigen- specific T cell from a human, for example, isolating of CD4 plus and CD8 plus from patient with prostate cancer or breast cancer.
Salam Alaikum Dear Shafieeh
I wonder what assay procedure you think about? FACS, IHC, or others?. As the answer depend on the assay you intend to follow.
Iltimas Duaa
Ali Mansoor
Isolating tumor specific T cells is tricky. People usually collect tumor tissues and isolate tumor infiltrating leukocytes. For you to be able to isolate antigen specific T cells, you need to know a specific marker that only the tumor specific T cells express. You could then use an antibody against that particular antigen to stain your cells and sort the +ve cells out. There are more complicated methods but all of them require you to somehow stain the tumor specific T cells with an antibody/tetramer that only the tumor specific T cells will bind to.
Article Isolation of neoantigen-specific T cells from tumor and peri...
If we are talking about tissues, you can process them by collagenase 4, DNAse 1 digestion followed by percol density. Here you will get CD4 and CD8 T cells ready to sort dor further analyze but you lack the antigen specificity information that you can only get by TCR sequencing (Adaptive company US) so you can select the most frequent TCR suppose to be specific for a define antigen regarding patients background.
On the other hand, if you already know the TCR sequence for your antigen, use tetramer/antibody to sort your cells of interest.
Dear Julie Joseph,
thanks for your reply.
but after isolation, i need tumor specific T cells to engineer and return them to the patient. so i am seeking a safe method.
I will definitely read that article that you attached.
Dear Ali Mansoor Al Ameri,
salam
I only want to know that is it possible to do such an isolation from a patient whereas they can be returned to the patient. because i searched about it but i could not find a good practical protocol.
Dear Fadi Jebbawi ,
i should think more about your answer and evaluate all its aspects. thank you
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