In short "no". Although we get very good staining of human basophils with anti-CCR3 using the LeukoSite clone, we could never get it to stain our allergen specific Th2 cells. That is, when we used 6h allergen activated PBMC and gated on CD4+, IL-4+ cells, we did not see these cells coexpressing CCR3. They did coexpress CRTH2 and CCR4. My general impression is that in follow-up, most investigators have not been able to demonstrate CCR3 as a Th2 marker on human T cells. The other issue that may be playing a role is downregulation of the receptor. For example, CRTH2 is downregulated in 7 day allergen specific Th2 lines, but is clearly expressed at earlier timepoints. So CCR3 could be on resting Th2 cells, but rapidly lost from the cells once activated and thus not present on your polyclonal Th2 lines. Calman -----Original Message----- From: Alessandro Serra [mailto:alessandro_serra@chiron.com] Sent: Saturday, May 06, 2006 5:55 PM To: cyto-inbox Subject: ccr3 on human th2 cells Dear All Has anybody looked at expression of ccr3 on polyclonal human th2 cell lines? the lines i made started from either cord blood or adult CD45RO- CD4+ tcells, they were cultured with IL-4 and anti-IFN-gamma and anti-IL-12. i can see upregulation of ccr4, crth2, cd30 but not ccr3. I am using the antibody from BD pharmingen which i tested on CD16- cells in the granulocyte morphological gate. These cells stained bright. Also i can stain a very small subsets of cd4+ cd3+ cells in the pbmcs taken from healthy adult individuals. So the antibody seems to work and very well. Has anybody been able to detect ccr3 expression in the conditions that i used and that are identical to those of the best papers? thanksReceived on Tue May 9 14:38:00 2006
This archive was generated by hypermail 2.1.8 : Wed May 10 2006 - 04:12:02 EDT
