Lymphocyte proliferation assay with specific antigen stimulation

Luck Felder luck_felder at hotmail.com
Tue Mar 30 09:18:38 EST 1999


Hello Hua-Chen Chang!

Hua-Chen Chang wrote:
> However, I have heard that live virus can also do the work.  The reason for
> inactivated virus for stimulation in lymphocyte proliferation is to provide a
> MHC class II processing pathway or avoid the cell killing by the virus, or any
> particular reason?
> 

You have to avoid cell killing, if You want to compare Your results
between different cell-probes with discret cell-numbers.

> Does MHC class II processing pathway is still required for in vitro lymphocyte
> proliferation assay?  CTL assay by standard Cr-51 release assayt does not need
> a live virus for stimulation.
> 
For antigen processing and presentation via MHC protein You don´t need
any living antigen. You need epitopes which can be recognized by Your
specific lymphocyte.

> Is APC required for nonspecific stimulation with ConA in this assay?

If ConA works as a mitogen, as it should, You don´t need APC´s, what
also means that You don´t need MHC II  processing for a proliferation
test.

> It only takes two days for ConA stimulation.  How many days does this assay
> need for antigen specific stimulation?

If You have a running system, You should always compare Your result out
of a new system to the results of Your running system. In my opinion,
You should start with 48h, which is a common time for proliferation
assays.

> 
> Thank you all.
> Hua-Chen Chang

Hope, helped You.

Yours sincerely,
LucK



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