Re: FACS naive mouse

From: Ondrej Hrusak (Ondrej.Hrusak@lfmotol.cuni.cz)
Date: Mon Oct 30 2000 - 06:34:17 EST


>    Hello, I am pretty much a FACS beginner. I have a  question, which
>nobody has been able to answer so far. Every time I do a FACS scan from
>peribronchial  lymph nodes (naive BALB/c mice) and stain it for cd3, cd4
>and cd8 cells I see a  huge population of cd4pos/cd3neg cells, as well as
>cd8pos/cd3neg cells. Does  anybody know what types of cells these are?  I
>have to admit that isolating PBLN >from naive mice  is rather tricky,
>usually leaving me with extracting the entire hilus and  meshing that.
>Could these cells derive from Thymus (which I am pretty sure to  extract
>beforehand) or from oesophagus? Thank you very much, Ph. Stock

In adidtion to the Hai Qi's comment, CD4+CD3- are also
monocytes/macrophages and CD8+CD3- are also some NK cells.
Both of them could be TCRgamma/delta+ or "true double negatives" (I mean
CD4-CD8-TCRalpha/beta+). I don't know what percentages you should expect in
your mouse tissue, though.
Good luck.
Ondrej

Ondrej Hrusak, M.D., Ph.D.
Institute of Immunology
Ondrej.Hrusak@lfmotol.cuni.cz
fax +4202 2443 5962
http://www.lf2.cuni.cz/clip/
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