Lymphocyte transformation by EBV - help wanted

[Cheung C. Yue]

It has been a while since I did EBV transformation.  Previously I had
used OKT3 (?to shut off autologous CTL response), but the last several
times I did this (in '89!!), I used cyclosporin as per a number of 
people's suggestions.  Well, the last 3 times I used cyclosporin I got
no transformed cell lines, but using OKT3, I had at least 1 out of 4
tries.  When I look at the cells, the OKT3 treated cultures developed
clustering of cells by the second day, and the media gets quite acidic
within a week.  In contrast there is not much cell clustering in the
cyclsporin treated cells even after 1 week.  Interestingly the cyclosporin
treated cells have on several occassions developed a bunch of large
granular-appearing cells which tended to stream together forming a
streak of cells (looks like a cotton fiber when out of focus, but is
actually a bunch of cells that line up in a tubular kind of structure).
Are these macrophages?  Have others observed this phenomenon?  Are the
clusters of cells in the OKT3-treated samples early transformed cells?
I kind of doubt the last point since the clusters do not have the same
appearance of the clusters of LCL, and they appear way too soon.

Anyone care to comment?  I would welcome getting in touch with anybody
on the net with experience transforming lymphocytes.  As it turns out,
nobody in Cleveland is actually doing lymphocyte transformation at present,
so I can't bug anybody local about this.  Thanks for all info.

C Cho Yue
ccy at po.cwru.edu
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