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During the course of in-vitro stimulation of both PBMCs and whole blood,
we invariably see down regulation of certain surface antigens (for
example, CD4 after 4 hours with PMA+Calcium Ionophore). These problems
vary depending on stimulus and incubation times. This obviously creates
problems as we try to identify populations following stimulation. This
leads to two questions:
1. Is anyone aware if these antigens are actually down-regulated (i.e.
internalized) or if they are shed into the supernatant?
2. As we use Brefeldin A to stop forward golgi transport (to trap
cytokines within the cell for detection with fluorochrome conjugated
Ab), would nocodazole, which blocks the return golgi pathway, work to
inhibit the internalization of surface antigen?
Thanks for any input you may have.
Keith Bahjat
Kbahjat@nwu.edu
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web