3 replies to this topic

[leelee]

Hi all

Does anyone know if it is possible to strip MHC class I molecules from the surface of cells (enzymatically, chemically, whatever...)?

I'm having a hard time finding any information on this, as every search I perform comes up with info on stripping the peptides from the mhc I molecules, but not the molecules themselves.

Fingers crossed one of you can help!

Thanks

leelee

[Astarte Biologics]

Low pH buffer (pH 3.0 citrate) can strip the peptides and beta 2 microglobulin off the surface.  The class  I heavy chain is not stable without the peptide and b2M so it is endocytosed.  But new molecules come to the surface all the time.  I used brefeldin A to stop transport and was able to see decreased class I on the surface by flow cytometry but they weren't negative, just shifted to the left.  

Hope that's helpful.

[leelee]

Very helpful, thank you!
I've been told something similar just recently by a colleague too, however I think they were a bit confused and so not very clear, meaning I had a hard time finding a paper or the like to verify it. Any chance you can point me in the right direction?

Thanks for the info re BfnA and the shifting of the population to the left. I think that will be ok for my purposes, I'm really just trying to explain a certain population of cells I'm getting rather than worrying about complete removal of MHC I from the surface.

[Astarte Biologics]

Search PubMed for the T2 cell line AND MHC class I and you should find some helpful papers.  They are back in the mid-1990's.  Let me know if you have any trouble tracking them down and I can give you specific papers.