On Fri, 16 May 1997 kevin-ault at uiowa.edu wrote:
> First of all, is lymphocyte proliferation testing still a well accepted
test to look at cell mediated responses?
Yes it is but it must be taken with a slight grain of salt. If the cells
proliferate in response to the antigen, they are accepted as being able to
recognize and proliferate in response to that antigen, but if they do not
this does not mean that they are not responsive to the antigen, just that
they do not proliferate. Other things that it is good to look at are:
do the cells produce T cell specific cytokines in reponse to the antigens?
are they cytotoxic to cells expressing the antigen.
It is a good idea to start with proliferation and move on from there. If
you have an idea of the expected response (intracellular pathogens
typically ellicite an TH1 reponse with IFN-gamma production) start from
there.
> Is priming with antigen or Il-2 for a week prior to actual testing a
common modification of the"traditional"lymphocyte proliferation testing?
Common, well that depends on what you are doing. Antigens that are weak
and have poor responses may need an expansion of the cells before usage.
(The priming expands or increases the number of precursors). If you are
looking for cytotoxicity this is done commonly however, if you are
attempting to find the number of precursors or whether an immuniztion
worked, no you would not want to do that. Look in the gold book (Current
protocols in Immunology) it gives a fair description of the assays and the
usage of the assays.
> Are the data generated (either stimulation indices or counts) normally
distrubuted?
I am not sure what you are asking here. They are relative to the assay
and antigen.
> Are non-radioactive methods a well accepted alternative to radiolabeled
> thymidine?
Yes they are, however, H3-thymidine is still the most common method. It
may take special equipment for either. (I perfer H3-thymidine)
TLWPhD
