Problem with Jurkat cells

From: Rana Nagarkatti (rana_nagarkatti@usa.net)
Date: Wed Jul 12 2000 - 06:49:10 EST


Dear Flowers,
 Hello and best wishes.
 I want to develop an assay for intracellular IL4 production in a human T cell
line. Currently I am using a Jurkat cell line. I have some doubts regarding
this cell line and would be grateful for any help in resolving these issues.
  1. There are a couple of Jurkat cell lines available from ATCC where it is
mentioned that they produce IL2. I would like to know if any one has used any
of these cell lines for looking at intracellular levels of IL4 after
stimulation with PMA and Ionomycin? JurkatE6.1?
 2. What are the cell surface markers which can define these cells? CD4, CD8
-ve is mentioned but what about IL2R, IL4R ?
 3. Are there any other human cell lines available which can be used for
detection of intracellular IL4, IL5 upon stimulationwith Ca2+ ionophores and
PMA.
 I am able to detect IL4 and IFN-g in PBMC which I use as a positive controll
for stimulation (though the conc of stimulants may not be the same for cell
lines- that is the only option I have), but have not been able to get any
signal from Jurkat cells. I have also tried increasing the amount of Ionomycin
and PMA but to no avail.
 Thanks and regards to all
 Rana Nagarkatti
 Center for Biochemical Technology
 Molecular Immunology Division
 Mall Road, Delhi, India 91-110007




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