I would like to investigate the T cells infiltration in several cells both in normal condition or due to cytokine/chemokine . May you help me to give some suggestion, the method to analysis the T cells infiltration both in normal cells and cancer cells and what is the parameter (Is it enough using CD4/CD8)?
Thanks
Hello
CD3,CD4,CD8 its good
For method you need pleas see all papers here , will help you .
for simultaneous localization and characterization of T cells in tissues use the novel method of multi epitope imaging with fluorochrome-tagged specific monoclonal antibodies .
Good Luck
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4337815/
http://www.nature.com/cmi/journal/v9/n4/full/cmi201215a.html
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2866629/
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3777488/
http://www.ncbi.nlm.nih.gov/pubmed/9820387
http://annonc.oxfordjournals.org/content/25/8/1536.full
http://www.sciencedirect.com/science/article/pii/S0165572813002348
https://books.google.com.sa/books?id=AzUmGT9F_wEC&pg=PA534&lpg=PA534&dq=method+to+analysis+the+T+cells+infiltration&source=bl&ots=FV8zspDL-t&sig=BMsb2EXOomTztxuHqgk-Kua4dcA&hl=ar&sa=X&ved=0ahUKEwjn2-yv2vfJAhXIUBQKHfu8DvwQ6AEIYDAJ#v=onepage&q=method%20to%20analysis%20the%20T%20cells%20infiltration&f=false
https://www.svi.edu.au/research_themes/human_t_cell
thank you saleh alkarim,
now, i want to investigate first using cell lines. your papers that you give to me almost using blood from cancer patients.
thanks
Hello, could you clarify your question - you mention using cell lines but what do you want to test being infiltrated? will it all be in vitro?
dear Olivia and Kamran, thanks, actually I want to investigate the mechanism of T cells infiltration in cancer cells. Previously study said that implanted cancer cells is causing infiltration of T cells. But I would like to look in vitro data to support it, both in normal cells and cancer cells, before I do the mutation in vivo.
yes I would like to look for the specific marker.
Dear Kamran Haq, yes. I would like to investigate several chemokine/cytokine. But I need some support data before I make the mutation on it. I try to implant several normal cells and cancer cells into mice, but I found several cells did not express or low express of CD8. So, I would like to investigate it. But making mutation in mice, quite need time. so, it is possible to analyze in cell line to support data exp. using a Magpie instrument? what kind of the parameter?
thanks
You have injected tumor cells in mice and you couldn't find immune cell infiltration, right? What kind of cells and mice did you use?
I think you want to determine in vitro whether a given tissue would be infiltrated with T cells. Then you should choose your cell lines of interest (normal and cancer) and do as Kamran said, look for surface markers expression and cytokine/chemokine production. With those tests you should get some information about the immunogenicity of you normal/cancer cells, but not about the actual immune cell infiltration.
You have to take into account that mimicking tumor microenvironment in vitro is not easy. You can try with 3D in vitro co-cultures using cancer, stromal and immune cell lines. This paper might be useful:
https://www.researchgate.net/publication/262268471_In_vitro_3D_models_of_tumor-immune_system_interaction
Hope this helps!
Article "In vitro" 3D models of tumor-immune system interaction
Thank you Raquel Mejias. yes, I injected LLC cells and Balb/c mice. Is it any possibility with the characteristic of the cells, such as highly metastases character? May I discuss with you about that paper?
Why did you choose an allogeneic model? I think it would be better to inject LLC cells in C57BL/6 mice. I don't have a lot of experience with 3D cultures, but you can send your questions and I'll do my best to help.
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