Re: FACS naive mouse

From: Ondrej Hrusak (Ondrej.Hrusak@lfmotol.cuni.cz)
Date: Tue Oct 31 2000 - 08:40:55 EST


Hello all,
As Mario Roederer reminded me, the second half of my suggestion was wrong.
I agree that TCR+ cells are always CD3+ and what I was talking about were
the CD3+CD4-CD8- (quite an opposite situation than what was observed in the
mouse tissue). Sorry for my confusion.

Ondrej


>>    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/
phone +4202 2443 5969, +4202 2443 2084
postal address:		V uvalu 84
			150 06 Praha 5
			Czech Republic



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