B7-H4 is a newly identified B7 homolog that plays an important role in maintaining T-cell homeostasis by inhibiting T-cell proliferation and lymphokine-secretion. In this study, we investigated the signal transduction pathways inhibited by B7-H4 engagement in mouse T cells. We found that treatment of CD3+ T cells with a B7-H4.Ig fusion protein inhibits anti-CD3 elicited T-cell receptor (TCR)/CD28 signaling events, including phosphorylation of the MAP kinases, ERK, p38, and JNK. B7-H4.Ig treatment also inhibited the phosphorylation of AKT kinase and impaired its kinase activity as assessed by the phosphorylation of its endogenous substrate GSK-3. Expression of IL-2 is also reduced by B7-H4. In contrast, the phosphorylation state of the TCR proximal tyrosine kinases ZAP70 and lymphocyte-specific protein tyrosine kinase (LCK) are not affected by B7-H4 ligation. These results indicate that B7-H4 inhibits T-cell proliferation and IL-2 production through interfering with activation of ERK, JNK, and AKT, but not of ZAP70 or LCK.
References
[1]
Sica GL, Choi IH, Zhu G, Tamada K, Wang SD, et al. (2003) B7-H4, a molecule of the B7 family, negatively regulates T cell immunity. Immunity 18: 849–861.
[2]
Prasad DV, Richards S, Mai XM, Dong C (2003) B7S1, a novel B7 family member that negatively regulates T cell activation. Immunity 18: 863–873.
[3]
Zang X, Loke P, Kim J, Murphy K, Waitz R, et al. (2003) B7x: a widely expressed B7 family member that inhibits T cell activation. Proc Natl Acad Sci U S A 100: 10388–10392.
[4]
Ou D, Wang X, Metzger DL, Ao Z, Pozzilli P, et al. (2006) Suppression of human T-cell responses to beta-cells by activation of B7-H4 pathway. Cell Transplant 15: 399–410.
[5]
Wang X, Hao J, Metzger DL, Mui A, Ao Z, et al. (2009) Local expression of B7-H4 by recombinant adenovirus transduction in mouse islets prolongs allograft survival. Transplantation 87: 482–490.
[6]
Weiss A, Imboden JB (1987) Cell surface molecules and early events involved in human T lymphocyte activation. Adv Immunol 41: 1–38.
[7]
Straus DB, Weiss A (1992) Genetic evidence for the involvement of the lck tyrosine kinase in signal transduction through the T cell antigen receptor. Cell 70: 585–593.
[8]
Irving BA, Chan AC, Weiss A (1993) Functional characterization of a signal transducing motif present in the T cell antigen receptor zeta chain. J Exp Med 177: 1093–1103.
[9]
Kolanus W, Romeo C, Seed B (1993) T cell activation by clustered tyrosine kinases. Cell 74: 171–183.
[10]
Whitehurst CE, Owaki H, Bruder JT, Rapp UR, Geppert TD (1995) The MEK kinase activity of the catalytic domain of RAF-1 is regulated independently of Ras binding in T cells. J Biol Chem 270: 5594–5599.
[11]
Calvo CR, Amsen D, Kruisbeek AM (1997) Cytotoxic T lymphocyte antigen 4 (CTLA-4) interferes with extracellular signal-regulated kinase (ERK) and Jun NH2-terminal kinase (JNK) activation, but does not affect phosphorylation of T cell receptor zeta and ZAP70. J Exp Med 186: 1645–1653.
Whitehurst CE, Geppert TD (1996) MEK1 and the extracellular signal-regulated kinases are required for the stimulation of IL-2 gene transcription in T cells. J Immunol 156: 1020–1029.
[14]
Owaki H, Varma R, Gillis B, Bruder JT, Rapp UR, et al. (1993) Raf-1 is required for T cell IL2 production. EMBO J 12: 4367–4373.
[15]
Su B, Jacinto E, Hibi M, Kallunki T, Karin M, et al. (1994) JNK is involved in signal integration during costimulation of T lymphocytes. Cell 77: 727–736.
[16]
Su B, Karin M (1996) Mitogen-activated protein kinase cascades and regulation of gene expression. Curr Opin Immunol 8: 402–411.
[17]
Cross DA, Alessi DR, Cohen P, Andjelkovich M, Hemmings BA (1995) Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature 378: 785–789.
[18]
Choi IH, Zhu G, Sica GL, Strome SE, Cheville JC, et al. (2003) Genomic organization and expression analysis of B7-H4, an immune inhibitory molecule of the B7 family. J Immunol 171: 4650–4654.
[19]
Yuan CL, Xu JF, Tong J, Yang H, He FR, et al. (2009) B7-H4 transfection prolongs beta-cell graft survival. Transpl Immunol 21: 143–149.
[20]
Tringler B, Zhuo S, Pilkington G, Torkko KC, Singh M, et al. (2005) B7-h4 is highly expressed in ductal and lobular breast cancer. Clin Cancer Res 11: 1842–1848.
[21]
Kryczek I, Zou L, Rodriguez P, Zhu G, Wei S, et al. (2006) B7-H4 expression identifies a novel suppressive macrophage population in human ovarian carcinoma. J Exp Med 203: 871–881.
[22]
Krambeck AE, Thompson RH, Dong H, Lohse CM, Park ES, et al. (2006) B7-H4 expression in renal cell carcinoma and tumor vasculature: associations with cancer progression and survival. Proc Natl Acad Sci U S A 103: 10391–10396.
[23]
Salceda S, Tang T, Kmet M, Munteanu A, Ghosh M, et al. (2005) The immunomodulatory protein B7-H4 is overexpressed in breast and ovarian cancers and promotes epithelial cell transformation. Exp Cell Res 306: 128–141.
[24]
Awadallah NS, Shroyer KR, Langer DA, Torkko KC, Chen YK, et al. (2008) Detection of B7-H4 and p53 in pancreatic cancer: potential role as a cytological diagnostic adjunct. Pancreas 36: 200–206.
[25]
Kryczek I, Wei S, Zou L, Zhu G, Mottram P, et al. (2006) Cutting edge: induction of B7-H4 on APCs through IL-10: novel suppressive mode for regulatory T cells. J Immunol 177: 40–44.
[26]
Toyooka K, Maruo S, Iwahori T, Yamamoto N, Tai XG, et al. (1996) CD28 co-stimulatory signals induce IL-2 receptor expression on antigen-stimulated virgin T cells by an IL-2-independent mechanism. Int Immunol 8: 159–169.
[27]
Schwartz RH (1990) A cell culture model for T lymphocyte clonal anergy. Science 248: 1349–1356.
[28]
Boise LH, Minn AJ, Noel PJ, June CH, Accavitti MA, et al. (1995) CD28 costimulation can promote T cell survival by enhancing the expression of Bcl-XL. Immunity 3: 87–98.
[29]
Meagher C, Tang Q, Fife BT, Bour-Jordan H, Wu J, et al. (2008) Spontaneous development of a pancreatic exocrine disease in CD28-deficient NOD mice. J Immunol 180: 7793–7803.
[30]
Honstettre A, Meghari S, Nunes JA, Lepidi H, Raoult D, et al. (2006) Role for the CD28 molecule in the control of Coxiella burnetii infection. Infect Immun 74: 1800–1808.
[31]
Parry RV, Reif K, Smith G, Sansom DM, Hemmings BA, et al. (1997) Ligation of the T cell co-stimulatory receptor CD28 activates the serine-threonine protein kinase protein kinase B. Eur J Immunol 27: 2495–2501.
[32]
Kane LP, Andres PG, Howland KC, Abbas AK, Weiss A (2001) Akt provides the CD28 costimulatory signal for up-regulation of IL-2 and IFN-gamma but not TH2 cytokines. Nat Immunol 2: 37–44.
[33]
Jones RG, Elford AR, Parsons MJ, Wu L, Krawczyk CM, et al. (2002) CD28-dependent activation of protein kinase B/Akt blocks Fas-mediated apoptosis by preventing death-inducing signaling complex assembly. J Exp Med 196: 335–348.
[34]
Van Parijs L, Refaeli Y, Lord JD, Nelson BH, Abbas AK, et al. (1999) Uncoupling IL-2 signals that regulate T cell proliferation, survival, and Fas-mediated activation-induced cell death. Immunity 11: 281–288.
[35]
Ahmed NN, Grimes HL, Bellacosa A, Chan TO, Tsichlis PN (1997) Transduction of interleukin-2 antiapoptotic and proliferative signals via Akt protein kinase. Proc Natl Acad Sci U S A 94: 3627–3632.
[36]
Kelly E, Won A, Refaeli Y, Van Parijs L (2002) IL-2 and related cytokines can promote T cell survival by activating AKT. J Immunol 168: 597–603.
[37]
Veillette A, Bookman MA, Horak EM, Bolen JB (1988) The CD4 and CD8 T cell surface antigens are associated with the internal membrane tyrosine-protein kinase p56lck. Cell 55: 301–308.
[38]
Karnitz L, Sutor SL, Torigoe T, Reed JC, Bell MP, et al. (1992) Effects of p56lck deficiency on the growth and cytolytic effector function of an interleukin-2-dependent cytotoxic T-cell line. Mol Cell Biol 12: 4521–4530.
[39]
Chiang GG, Sefton BM (2000) Phosphorylation of a Src kinase at the autophosphorylation site in the absence of Src kinase activity. J Biol Chem 275: 6055–6058.
[40]
Kong G, Dalton M, Bubeck Wardenburg J, Straus D, Kurosaki T, et al. (1996) Distinct tyrosine phosphorylation sites in ZAP-70 mediate activation and negative regulation of antigen receptor function. Mol Cell Biol 16: 5026–5035.
