Abstract
A coordinated effort combining bioinformatic tools with high-throughput cell-based screening assays was implemented to identify novel factors involved in T-cell biology. We generated a unique library of cDNAs encoding predicted secreted and transmembrane domain–containing proteins generated by analyzing the Human Genome Sciences cDNA database with a combination of two algorithms that predict signal peptides. Supernatants from mammalian cells transiently transfected with this library were incubated with primary T cells and T-cell lines in several high-throughput assays. Here we describe the discovery of a T cell factor, TIP (T cell immunomodulatory protein), which does not show any homology to proteins with known function. Treatment of primary human and murine T cells with TIP in vitro resulted in the secretion of IFN-γ, TNF-α, and IL-10, whereas in vivo TIP had a protective effect in a mouse acute graft-versus-host disease (GVHD) model. Therefore, combining functional genomics with high-throughput cell-based screening is a valuable and efficient approach to identifying immunomodulatory activities for novel proteins.
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References
Hertzberg, R.P. & Pope, A.J. High-throughput screening: new technology for the 21st century. Curr. Opin. Chem. Biol. 4, 445–451 (2000).
Fox, S., Farr-Jones, S. & Yund, M.A. High-throughput screening for drug discovery: continually transitioning into new technology. J. Biomol. Screen. 4, 183–186 (1999).
Eddy, S.R. Profile hidden Markov models. Bioinformatics 14, 755–763 (1998).
Nielsen, H., Engelbrecht, J., Brunak, S. & von Heijne, G. Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng. 10, 1–6 (1997).
Barash, S., Wei, W., & Shi, Y. Human secretory signal peptide description by hidden Markov model and generation of a strong artificial signal peptide for secreted protein expression. Biochem. Biophys. Res. Commun. 224, 835–842 (2002).
Chen, C. et al. An integral functional genomics screening program reveals novel a role for BMP-9 in glucose homeostasis. Nat. Biotechnol. 21, 294–301 (2003).
Swartzman, E.E., Miraglia, SJ., Mellentin-Michelott, J., Evangelista, L. & Yuan, P.M. Homogeneous cell- and bead-based assays for high-throughput screening using fluorometric microvolume assay technology. J. Biomol. Screen. 4, 193–204 (1999).
Gurlo, T., Huang, W.-W. & von Grafstein, H. PGE2 inhibits IL-2- and IL-4-dependent proliferation of CTLL-2 and HT2 cells. Cytokine 10, 265–274 (1998).
Latchman, Y. et al. PD-L2 is a second ligand for PD-1 and inhibits T-cell activation. Nat. Immunol. 2, 261–268 (2001).
Tseng, S.Y. et al. B7-DC, a new dendritic cell molecule with potent costimulatory properties for T cells. J. Exp. Med. 193, 839–846 (2001).
Freeman, G.J. et al. Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. J. Exp. Med. 192, 1027–1034 (2000).
Yang, Y.G., Dey, B.R., Sergio, J.J., Perason, D.A. & Sykes, M. Donor-derived interferon-γ is required for inhibition of acute graft-versus-host disease by interleukin 12. J. Clin. Invest. 102, 2126–2135 (1998).
Cho, C.Q., Wittmer, S. & Dalton, D.K. Failure to suppress the expansion of the activated CD4 T-cell population in interferon-γ-deficient mice leads to exacerbation of experimental autoimmune encephalomyelitis. J. Exp. Med. 192, 123–128 (2000).
Brok, H.P., Vossen, J.M. & Heidt, P.J. IFN-γ-mediated prevention of graft-versus-host-disease: pharmacodynamic studies and influence on proliferative capacity of chimeric spleen cells. Bone Marrow Transplant. 22, 1005–1010 (1998).
Holler, E. et al. Prognostic significance of increased IL-10 production in patients prior to allogeneic bone marrow transplantation. Bone Marrow Transplant. 25, 237–241 (2000).
LaFleur, D.W. et al. Interferon-κ, a novel type I interferon expressed in human keratinocytes. J. Biol. Chem. 276, 39765–39771 (2001).
Forrer, P., Tamaskovic, R. & Jaussi, R. Enzyme-linked immunosorbent assay for measurement of JNK, ERK, and p38 kinase activities. Biol. Chem. 379, 1101–1110 (1998).
Kotanides, H. & Reich, N.C. Requirement of tyrosine phosphorylation for rapid activation of DNA binding factor by IL-4. Science 262, 1265–1267 (1993).
Gleichmann, E., Pals, S.T., Rolink, A.G., Radaszkiewicz, T. & Gleichmann, H. Graft-versus-host reactions: clues to the etiopathology of a spectrum of immunological disease. Immunol. Today 5, 324–332 (1984).
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The authors wish to thank Andrea Smith for assistance with the manuscript preparation.
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Fiscella, M., Perry, J., Teng, B. et al. TIP, a T-cell factor identified using high-throughput screening increases survival in a graft-versus-host disease model. Nat Biotechnol 21, 302–307 (2003). https://doi.org/10.1038/nbt797
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DOI: https://doi.org/10.1038/nbt797
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