Epi-allelic Erk1 and Erk2 knockdown series for quantitative analysis of T cell Erk regulation and IL-2 production.

TitleEpi-allelic Erk1 and Erk2 knockdown series for quantitative analysis of T cell Erk regulation and IL-2 production.
Publication TypeJournal Article
Year of Publication2007
AuthorsWille, L, Kemp, ML, Sandy, P, Lewis, CL, Lauffenburger, DA
JournalMol Immunol
Volume44
Issue12
Pagination3085-91
Date Published2007 May
ISSN0161-5890
KeywordsAlleles, Animals, Cell Line, Hybridomas, Interleukin-2, Lymphocyte Activation, Methods, Mice, Mitogen-Activated Protein Kinase 1, Mitogen-Activated Protein Kinase 3, Phosphorylation, RNA, Small Interfering, T-Lymphocytes
Abstract

Erk activation is often used as a downstream pathway indicator of TCR signaling, generally in terms of both Erk1 and Erk2 isoforms measured together. In order to investigate potential distinctions between Erk1 and Erk2 regulation and effects downstream of TCR ligation, we generated a series of stable and independent Erk1 and Erk2 shRNA knockdown lines in the 1B6 T cell hybridoma. We observed no compensatory effect by opposite isoform upregulation, and found similar fractions of total phosphorylated Erk1/2 across this epi-allelic series in response to both anti-CD3 and peptide-MHC stimulation of TCR. Moreover, a previous prediction of an isoform-independent linear relationship between Erk activation and IL-2 production was confirmed. The effect of the shRNA-mediated knockdowns in reducing IL-2 production was observed to be stronger than that arising from pharmacological MEK inhibition at comparable degrees of ERK1/2 phosphorylation levels.

DOI10.1016/j.molimm.2007.02.008
Alternate JournalMol. Immunol.
PubMed ID17418417
PubMed Central IDPMC2692299
Grant ListP50 GM068762-060002 / GM / NIGMS NIH HHS / United States
P50-GM68762 / GM / NIGMS NIH HHS / United States
R01-AI65824 / AI / NIAID NIH HHS / United States
U54-CA112967 / CA / NCI NIH HHS / United States