A cytokine promoter/yellow fluorescent protein reporter transgene serves as an early activation marker of lymphocyte subsets.
A mouse containing an IL-4 promoter linked to the yellow fluorescent protein (YFP) reporter transgene was created to follow aspects of lymphocyte development and function. Following stimulation with phorbol 12-myristate 13-acetate and ionomycin, anti-CD3/CD28, antigen-specific peptide, or allogeneic cells, both CD4 and CD8 T cells expressed the transgene within 24h in a manner that was consistent with cellular activation markers. Transgene induction was inhibited by cyclosporine and FK506, suggesting that its activation occurs in an NFAT-dependent manner. B lymphocytes were also able to express the transgene when stimulated with LPS. This induction was inhibited in part by rapamycin. The results suggest that this transgene can function as an indicator of lymphocyte activation. Because YFP is not toxic and requires no preparation of the cells to view the reporter gene, this system provides a unique tool to follow lymphocyte activation in a number of model systems, such as those involving transplantation, allergy, and vaccine development.
Kumar
Shantha
S
Department of Microbiology and Immunology, Emory University School of Medicine, 1510 Clifton Road, Atlanta, GA 30322, USA.
Skeen
Marianne J
MJ
Adiri
Yaffa
Y
Yoon
Hyseuk
H
Vezys
Vaiva D
VD
Lukacher
Aron E
AE
Evavold
Brian D
BD
Ziegler
H Kirk
HK
Boss
Jeremy M
JM
D000818:Animals; D000941:Antigens / Q000008:administration & dosage; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D015415:Biomarkers; D016572:Cyclosporine / Q000494:pharmacology; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D066298:In Vitro Techniques; D015847:Interleukin-4 / Q000235:genetics*; D015759:Ionomycin / Q000494:pharmacology; D008164:Luminescent Proteins / Q000235:genetics*; D008213:Lymphocyte Activation; D016131:Lymphocyte Subsets / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010047:Ovalbumin / Q000008:administration & dosage / Q000276:immunology; D010446:Peptide Fragments / Q000008:administration & dosage / Q000276:immunology; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics; D016559:Tacrolimus / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000941:Antigens; D001426:Bacterial Proteins; D015415:Biomarkers; D004274:DNA, Recombinant; D008164:Luminescent Proteins; C111390:OVA 323-339; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C054562:yellow fluorescent protein, Bacteria; D015847:Interleukin-4; D015759:Ionomycin; D016572:Cyclosporine; D010047:Ovalbumin; D013755:Tetradecanoylphorbol Acetate; D016559:Tacrolimus