T cell proliferation

Tom Thatcher ttha at uhura.cc.rochester.edu
Thu Jan 9 10:12:40 EST 1997


In article <5b249k$cu5 at mserv1.dl.ac.uk>,
Ashok M S  <lambu at biochem.iisc.ernet.in> wrote:
>
>Hi,        i am tryingto look for antigen specfic t cell proliferation
>    but my protein get solubilised in 8M urea or guanidine hydrochloride.
>I have tried electroelution, dialysized aganist 30-50% glycerol as my
>protein doesnot ppt, yet i see no proliferation. I have also tried 
>using inactivated virus as anitgen, again there was no proliferation.
>can you help me.
>
>thank you
>
>Ashok.M.S
>dept of biochemistry
>indian institute of science
>bangalore 12
>e-mail lambu at biochem.iisc.ernet.in

Are these T cells from immunized experimental animals (like mice) or
PBLs from human subjects?  How do you know that the T cells you are
using *should* proliferate?

Take your protein dissolved in urea and dialyse against water or PBS.
You said it remains soluble.  Then precipitate it with acetone or
evaporate the water.  Try immunizing some mice with the protein then
testing T cell proliferation with the same preparation to make sure the
purification method you used keeps the protein in an antigenic state.
In other words, make sure this protein preparation works in a favorable
system before returning to the system in which it doesn't work.

For the virus study, how did you inactivate the virus?  Did you add
antigen presenting cells to your T cells so they could break down
the virus and present epitopes?

-- 
Tom Thatcher                          | You can give a PC to a Homo habilis,
University of Rochester Cancer Center | and he'll use it, but he'll use it
ttha at uhura.cc.rochester.edu           | to crack nuts.



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