Identification of genes differentially expressed in T cells following stimulation with the chemokines CXCL12 and CXCL10.
BACKGROUND
Chemokines are involved in many biological activities ranging from leukocyte differentiation to neuronal morphogenesis. Despite numerous reports describing chemokine function, little is known about the molecular changes induced by cytokines.


METHODS
We have isolated and identified by differential display analysis 182 differentially expressed cDNAs from CXCR3-transfected Jurkat T cells following treatment with CXCL12 or CXCL10. These chemokine-modulated genes were further verified using quantitative RT-PCR and Western blot analysis.


RESULTS
One hundred and forty-six of the cDNAs were successfully cloned, sequenced, and identified by BLAST. Following removal of redundant and non-informative clones, seventeen mRNAs were found to be differentially expressed post treatment with either chemokine ligand with several representing known genes with established functions. Twenty-one genes were upregulated in these transfected Jurkat cells following both CXCL12 and CXCL10, four genes displayed a discordant response and seven genes were downregulated upon treatment with either chemokine. Identified genes include geminin (GEM), thioredoxin (TXN), DEAD/H box polypeptide 1 (DDX1), growth hormone inducible transmembrane protein (GHITM), and transcription elongation regulator 1 (TCERG1). Subsequent analysis of several of these genes using semi-quantitative PCR and western blot analysis confirmed their differential expression post ligand treatment.


CONCLUSIONS
Together, these results provide insight into chemokine-induced gene activation and identify potentially novel functions for known genes in chemokine biology.
Clinical Immunology Section, Laboratory of Immunology, Gerontology Research Center, National Institute on Aging, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224, USA. wichita3@comcast.net
J E
JE
Nagel
R J
RJ
Smith
L
L
Shaw
D
D
Bertak
V D
VD
Dixit
E M
EM
Schaffer
D D
DD
Taub
D015153:Blotting, Western; D020013:Calcium Signaling / Q000187:drug effects; D054357:Chemokine CXCL10; D054377:Chemokine CXCL12; D019743:Chemokines, CXC / Q000494:pharmacology*; D002633:Chemotaxis / Q000187:drug effects; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D006801:Humans; D019169:Jurkat Cells / Q000187:drug effects / Q000378:metabolism; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D054367:Receptors, CXCR3; D019718:Receptors, CXCR4 / Q000187:drug effects; D019707:Receptors, Chemokine / Q000187:drug effects / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013382:Subtraction Technique; D013601:T-Lymphocytes / Q000187:drug effects* / Q000378:metabolism; D015533:Transcriptional Activation; D014162:Transfection
C515255:CXCL12 protein, human; C515187:CXCR3 protein, human; D054357:Chemokine CXCL10; D054377:Chemokine CXCL12; D019743:Chemokines, CXC; D018076:DNA, Complementary; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D054367:Receptors, CXCR3; D019718:Receptors, CXCR4; D019707:Receptors, Chemokine; D011993:Recombinant Fusion Proteins