> -----Original Message----- > From: David Ritchie [SMTP:dritchie@malaghan.org.nz] > Sent: Thursday, 17 October 2002 4:18 AM > To: Cytometry Mailing List > Subject: CD4 on stimulated human T cells > > David, > > This is a well recognised phemomenom with CD4 and CD56 (and CD14 if LPS > concentrations are too high). Dont know whether these molecules are lost > or some internalised. CD4 is usually brighter if you stain surface and > cytokines following permeabilisation, so some CD4 may be internalised. > Most workers get around the problem by analysing the CD3+/CD8- (CD4+) > subset but you should check that CD3/CD8 + CD3/CD4 = total CD3 cells in > case of CD3+/CD8-/CD4- cells present in sample. > > Greg Hodge > > > Any suggestions welcomed on below problem, > > We are tracking the cytokine profiles (by intracellular staining) produced > by CD4+ T cells in patients who have undergone allogeneic bone marrow > transplant (BMT). We are looking at cytokine production pre- and > post-stimulation of T cells with PHA/ionomycin for 4 hours. We have no > difficulty clearly identifying CD4+ T cells on the unstimulated sample, > but > on the stimulated sample CD4 seems to disappear. CD3 is still clearly > evident. Has anyone seen this before? Is the cell fixing process used for > ic > staining likely to interfere with CD4 expression or anti-CD4 binding? We > don't seem to be losing cells in the short culture period. > > Thanks for any comments > > David Ritchie
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