Dear everybody,
Plz help me, Now I am doing in vivo mouse tumor model by using dendritic cells combined with v-FlaB (as adjuvant) against colon cancer model (MC-38 cells). but some papers mention, MC-38 cells don't express TLR-5, and also I am using the conventional DC (that's maybe also don't appear TLR5 on their surface).
Now I am finding the hypothesis of DC+ FlaB in vivo for mouse model, so may I check the expression of TLR-5 on or in MC-38 by FACs analysis or by confocol microscopy, or do you have another explanation of the hypothesis: "DC+ FlaB against MC-38 colon cancer in vivo mouse model".
Thank you sincerely,
Yes you can check the expression of TLRs at the cell membrane by FACS or immunofluorescence or by western blotting to see if its expressed in the intra-cellular compartement
Hello
Mohammed has given you a good answer
http://jnumedmtg.snmjournals.org/cgi/content/short/55/1_MeetingAbstracts/171
Hi !
I would go for Q-PCR. You could use CT26.WT as a negative control. As shown by Next Generation Sequencing, these bugs are not expressing Tlr5.
You can find this NGS data for CT26.WT in the supplementary table of Castle et al.:
http://www.biomedcentral.com/1471-2164/15/190
But indeed, as you basically pointed out, it is the DC which need to express Tlr5 for your experiment (which is most likely the case). If the MC-38 also express Tlr5, this would rather not lead to an immune response, but potentially to an activation of NFkB via Myd88 in these cells. And this could -theoretically- increase treatment resistance via upregulation of anti apoptotic molecules (Bcl-2, Bcl-XL, Mcl-1, Survivin...). So, conclusively, what should probably better not be done would be to inject the adjuvant into the tumor, if it is Tlr5 positive.
In any case, i wish you good luck with your experiment ! :-)
Chees,
Norman
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