A cytokine promoter/yellow fluorescent protein reporter transgene serves as an early activation marker of lymphocyte subsets

• Published in: Cellular Immunology Vol. 237; no. 2; pp. 131 - 140
• Main Authors: Kumar, Shantha, Skeen, Marianne J., Adiri, Yaffa, Yoon, Hyseuk, Vezys, Vaiva D., Lukacher, Aron E., Evavold, Brian D., Ziegler, H. Kirk, Boss, Jeremy M.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01.10.2005
• Subjects: Animals; Antigens - administration & dosage; Bacterial Proteins - genetics; Base Sequence; Biomarkers; CD4/CD8 cells; Cyclosporine - pharmacology; DNA, Recombinant - genetics; Gene Expression - drug effects; Genes, Reporter; In Vitro Techniques; Interleukin-4 - genetics; Ionomycin - pharmacology; Luminescent Proteins - genetics; Lymphocyte Activation; Lymphocyte Subsets - drug effects; Lymphocyte Subsets - immunology; Lymphocyte Subsets - metabolism; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin - administration & dosage; Ovalbumin - immunology; Peptide Fragments - administration & dosage; Peptide Fragments - immunology; Promoter Regions, Genetic; Recombinant Fusion Proteins - genetics; T cell activation; Tacrolimus - pharmacology; Tetradecanoylphorbol Acetate - pharmacology; Transgenic; T cell activation; Transgenic; CD4/CD8 cells
• ISSN: 0008-8749; 1090-2163; 1365-2567
• DOI: 10.1016/j.cellimm.2005.11.003

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 24 h 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.