HL-60/Pneumococcal opsonophagocytosis assay

From: Bryant, Jenny (NSW) (JBryant@arcbs.redcross.org.au)
Date: Wed Dec 13 2000 - 00:53:01 EST


Hi,

We have been working on an opsonophagocytosis assay to detect
anti-Pneumococcal antibodies in serum using DMF differentiated HL-60 cells
as the effector cells.  We really have read many papers on this and related
subjects.  However, we have only found passing references to the problems
which we are encountering.  We have also found some confusion in published
methods where the same authors use very different methods over a number of
papers.

We are finding phagocytosis in the presence of complement only (baby rabbit
serum).  Different batches have been tried.

How long can you passage HL-60 cells and expect them (after differentiation)
to still be useable in such an assay?

Is anyone running assays like this and could offer some advice on methods?
After a particularly frustrating few weeks, we would be grateful for any
help on this matter.

Jenny

Jenny Bryant
Flow Cytometry
Australian Red Cross Blood Service - NSW/ACT
153 Clarence St
Sydney, NSW 2000, AUSTRALIA     .._|\
Ph:     61 2 9229 4341              /      \
FAX:    61 2 9229 4521             \_.-._/<<<<<<
E-mail:  jbryant@arcbs.redcross.org.au   v



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