Fyn Expression Predicates Both Protective Immunity and Onset of Autoimmunity

Behrouz Moemeni^1,2, Nathalie Vacaresse¹, Marina Vainder¹, Michael E. Wortzman², Tania H. Watts², André Veillette³, Thomas F. Gajewski⁴, Michael Julius^1,2

¹Sunnybrook Research Institute, Toronto, Canada.
²Department of Immunology, University of Toronto, Toronto, Canada.
³Laboratory of Molecular Oncology, Clinical Research Institute of Montreal, Montreal, Canada.
⁴Department of Pathology, University of Chicago, Chicago, IL, USA.

DOI: 10.4236/oji.2015.55020                

Abstract

Genomic disruption of Fyn has not been associated with an immune-deficient phenotype, notwithstanding the profound impairment in IL-2 production by T cells derived from Fyn-deficient animals observed in vitro. The results presented demonstrate that Fyn deficient animals succumb to influenza infection ahead of the protective expansion of lung infiltrating T cells and viral clearance observed in wild-type hosts. Formal proof that Fyn-dependent IL-2 production mediates T cell expansion in vivo is provided using a model of T cell induced enteropathy. Specifically, Fyn deficient naive T cells do not induce colitis in SCID animals due to their lack of expansion, and Fyn re-expression rescues both IL-2 production and its capacity to support in vivo expansion leading to colitis. These results reconcile the obligatory role of Fyn in T cell activation and autocrine IL-2 supported growth; and underscore the mechanism through which its function is integrated with and regulated by Lck.

Keywords

Fyn, Lck, TcR Signaling, Immunity, Autoimmunity

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Appleby, M.W., Gross, J.A., Cooke, M.P., Levin, S.D., Qian, X. and Perlmutter, R.M. (1992) Defective T Cell Receptor Signaling in Mice Lacking the Thymic Isoform of p59fyn. Cell, 70, 751-763. http://dx.doi.org/10.1016/0092-8674(92)90309-Z
[2]	Stein, P.L., Lee, H., Rich, S. and Soriano, P. (1992) pp59fyn Mutant Mice Display Differential Signaling in Thymocytes and Peripheral T Cells. Cell, 70, 741-750. http://dx.doi.org/10.1016/0092-8674(92)90308-Y
[3]	Mamchak, A.A., Sullivan, B.M., Hou, B., Lee, L.M., Gilden, J.K., Krummel, M.F., Locksley, R.M. and DeFranco, A.L. (2008) Normal Development and Activation but Altered Cytokine Production of Fyn-Deficient CD4+ T Cells. The Journal of Immunology, 181, 5374-5385. http://dx.doi.org/10.4049/jimmunol.181.8.5374
[4]	Eberl, G., Lowin-Kropf, B. and MacDonald, H.R. (1999) Cutting Edge: NKT cell Development Is Selectively Impaired in Fyn-Deficient Mice. The Journal of Immunology, 163, 4091-4094.
[5]	Filipp, D., Leung, B.L., Zhang, J., Veillette, A. and Julius, M. (2004) Enrichment of LCK in Lipid Rafts Regulates Colocalized Fyn Activation and the Initiation of Proximal Signals through TCR Alpha Beta. The Journal of Immunology, 172, 4266-4274. http://dx.doi.org/10.4049/jimmunol.172.7.4266
[6]	Filipp, D., Zhang, J., Leung, B.L., Shaw, A., Levin, S.D., Veillette, A. and Julius, M. (2003) Regulation of Fyn through Translocation of Activated LCK into Lipid Rafts. The Journal of Experimental Medicine, 197, 1221-1227. http://dx.doi.org/10.1084/jem.20022112
[7]	Filipp, D., Moemeni, B., Ferzoco, A., Kathirkamathamby, K., Zhang, J., Ballek, O., Davidson, D., Veillette, A. and Julius, M. (2008) Lck-Dependent Fyn Activation Requires C-Terminus-Dependent Targeting of Kinase-Active Lck to Lipid Rafts. Journal of Biological Chemistry, 283, 26409-26422. http://dx.doi.org/10.1074/jbc.M710372200
[8]	Leach, M.W., Bean, A.G., Mauze, S., Coffman, R.L. and Powrie, F. (1996) Inflammatory Bowel Disease in C.B-17 Scid Mice Reconstituted with the CD45RB High Subset of CD4+ T Cells. American Journal of Pathology, 148, 1503- 1515.
[9]	Powrie, F., Leach, M.W., Mauze, S., Caddie, L.B. and Coffman, R.L. (1993) Phenotypically Distinct Subsets of CD4+ T Cells Induce or Protect FROM Chronic Intestinal Inflammation in C. B-17 Scid Mice. International Immunology, 5, 1461-1471. http://dx.doi.org/10.1093/intimm/5.11.1461
[10]	Powrie, F., Mauze, S. and Coffman, R.L. (1997) CD4+ T Cells in the Regulation of Inflammatory Responses in the Intestine. Research in Immunology, 148, 576-581. http://dx.doi.org/10.1016/S0923-2494(98)80152-1
[11]	Ennis, F.A., Verbonitz, M., Reichelderfer, P. and Daniel, S. (1976) Recombination of Influenza A Virus Strains: Effect on Pathogenicity. Developments in Biological Standardization, 33, 220-225.
[12]	Lin, G.H., Sedgmen, B.J., Moraes, T.J., Snell, L.M., Topham, D.J. and Watts, T.H. (2009) Endogenous 4-1BB Ligand Plays a Critical Role in Protection from Influenza-Induced Disease. Journal of immunology, 182, 934-947. http://dx.doi.org/10.4049/jimmunol.182.2.934
[13]	Wan, Y.Y., Leon, R.P., Marks, R., Cham, C.M., Schaack, J., Gejewski, T.F. and Degregori, J. (2000) Transgenic Expression of the Coxasackie/Adenovirus Receptor Enables Adenoviral-Mediated Gene Delivery in Naive T Cells. Proceedings of the National Academy of Sciences of the United States of America, 97, 13784-13789. http://dx.doi.org/10.1073/pnas.250356297
[14]	Cottey, R., Rowe, C.A. and Bender, B.S. (2001) Influenza Virus. In: Coico, R., Ed., Current Protocols in Immunology, John Wiley and Sons, Hoboken. http://dx.doi.org/10.1002/0471142735.im1911s42
[15]	Poussier, P., Ning, T., Chen, J., Banerjee, D. and Julius, M. (2000) Intestinal Inflammation Observed in IL-2R/IL-2 Mutant Mice Is Associated with Impaired Intestinal T Lymphopoiesis. Gastroenterology, 118, 880-891. http://dx.doi.org/10.1016/S0016-5085(00)70174-0
[16]	Quah, B.J., Warren, H.S. and Parish, C.R. (2007) Monitoring Lymphocyte Proliferation in Vitro and in Vivo with the Intracellular Fluorescent Dye Carboxyfluorescein Diacetate Succinimidyl Ester. Nature Protocols, 2, 2049-2056. http://dx.doi.org/10.1038/nprot.2007.296
[17]	Marmour, M.D. and Julius, M. (2001) Role for Lipid Rafts in Regulating Interleukin-2 Receptor Signaling. Blood, 98, 1489-1497. http://dx.doi.org/10.1182/blood.V98.5.1489
[18]	Filby, A., Seddon, B., Kleczkowska, J., Salmond, R., Tomlinson, P., Smida, M., Lindquist, J.A., Schraven, B. and Zamoyska, R. (2007) Fyn Regulates the Duration of TCR Engagement Needed for Commitment to Effector Function. Journal of Immunology, 179, 4635-4644. http://dx.doi.org/10.4049/jimmunol.179.7.4635
[19]	Sugie, K., Jeon, M. and Grey, H.M. (2004) Activation of Naive CD4 T Cells by Anti-CD3 Reveals an Important Role for Fyn in Lck-Mediated Signaling. Proceedings of the National Academy of Sciences of the United States of America, 101, 14859-14864. http://dx.doi.org/10.1073/pnas.0406168101
[20]	da Silva, A., Rosenfield, J., Mueller, I., Bouton, A., Hirai, H. and Rudd, C. (1997) Biochemical Analysis of p120/130: A Protein-Tyrosine Kinase Substrate Restricted to T and Myeloid Cells. The Journal of Immunology, 158, 2007-2016.
[21]	Bettelli, E., Carrier, Y., Gao, W., Korn, T., Strom, T.B., Oukka, M., Weiner, H.L. and Kuchroo, V.K. (2006) Reciprocal Developmental Pathways for the Generation of Pathogenic Effector TH17 and Regulatory T Cells. Nature, 441, 235- 238. http://dx.doi.org/10.1038/nature04753
[22]	Veldhoen, M., Hocking, R.J., Atkins, C.J., Locksley, R.M. and Stockinger, B. (2006) TGFβ in the Context of an Inflammatory Cytokine Milieu Supports de Novo Differentiation of IL-17-Producing T Cells. Immunity, 24, 179-189. http://dx.doi.org/10.1016/j.immuni.2006.01.001
[23]	Khanolkar, A., Fuller, M.J. and Zajac, A.J. (2004) CD4 T Cell-Dependent CD8 T Cell Maturation. Journal of Immunology, 172, 2834-2844. http://dx.doi.org/10.4049/jimmunol.172.5.2834
[24]	Williams, M.A., Tyznik, A.J. and Bevan, M.J. (2006) Interleukin-2 Signals during Priming Are Required for Secondary Expansion of CD8+ Memory T Cells. Nature, 441, 890-893. http://dx.doi.org/10.1038/nature04790
[25]	Wilson, E.B. and Livingstone, A.M. (2008) Cutting Edge: CD4+ T Cell-Derived IL-2 Is Essential for Help-Dependent Primary CD8+ T Cell Responses. Journal of Immunology, 181, 7445-7448. http://dx.doi.org/10.4049/jimmunol.181.11.7445
[26]	Thomas, P.G., Keating, R., Hulse-Post, D.J. and Doherty, P.C. (2006) Cell-Mediated Protection in Influenza Infection. Emerging Infectious Diseases, 12, 48-54. http://dx.doi.org/10.3201/eid1201.051237
[27]	Gillis, S., Ferm, M.M., Ou, W. and Smith, K.A. (1978) T Cell Growth Factor: Parameters of Production and a Quantitative Microassay for Activity. Journal of Immunology, 120, 2027-2032.
[28]	Robb, R.J. and Smith, K.A. (1981) Heterogeneity of Human T-Cell Growth Factor(s) Due to Variable Glycosylation. Molecular Immunology, 18, 1087-1094. http://dx.doi.org/10.1016/0161-5890(81)90024-9
[29]	Smith, K.A. (1988) Interleukin-2: Inception, Impact, and Implications. Science, 240, 1169-1176. http://dx.doi.org/10.1126/science.3131876
[30]	Sojka, D.K., Bruniquel, D., Schwartz, R.H. and Singh, N.J. (2004) IL-2 Secretion by CD4+ T Cells in Vivo Is Rapid, Transient, and Influenced by TCR-Specific Competition. Journal of Immunology, 172, 6136-6143. http://dx.doi.org/10.4049/jimmunol.172.10.6136
[31]	Sadlack, B., Merz, H., Schorle, H., Schimpl, A., Feller, A.C. and Horak, I. (1993) Ulcerative Colitis-Like Disease in Mice with a Disrupted Interleukin-2 Gene. Cell, 75, 253-261. http://dx.doi.org/10.1016/0092-8674(93)80067-O
[32]	Willerford, D.M., Chen, J., Ferry, J.A., Davidson, L., Ma, A. and Alt, F.W. (1995) Interleukin-2 Receptor Alpha Chain Regulates the Size and Content of the Peripheral Lymphoid Compartment. Immunity, 3, 521-530. http://dx.doi.org/10.1016/1074-7613(95)90180-9
[33]	Schorle, H., Holtschke, T., Hunig, T., Schimpl, A. and Horak, I. (1991) Development and Function of T Cells in Mice Rendered Interleukin-2 Deficient by Gene Targeting. Nature, 352, 621-624. http://dx.doi.org/10.1038/352621a0
[34]	Poussier, P., Ning, T., Banerjee, D. and Julius, M. (2002) A Unique Subset of Self-Specific Intraintestinal T Cells Maintains Gut Integrity. Journal of Experimental Medicine, 195, 1491-1497. http://dx.doi.org/10.1084/jem.20011793
[35]	Papiernik, M., de Moraes, M.L., Pontoux, C., Vasseur, F. and Penit, C. (1998) Regulatory CD4 T Cells: Expression of IL-2R Alpha Chain, Resistance to Clonal Deletion and IL-2 Dependency. International Immunology, 10, 371-378. http://dx.doi.org/10.1093/intimm/10.4.371
[36]	Kameyama, K., Nemoto, Y., Kanai, T., Shinohara, T., Okamoto, R., Tsuchiya, K., Nakamura, T., Sakamoto, N., Totsuka, T., Hibi, T. and Watanabe, M. (2010) IL-2 Is Positively Involved in the Development of Colitogenic CD4+ IL-7R Alpha High Memory T Cells in Chronic Colitis. European Journal of Immunology, 40, 2423-2436. http://dx.doi.org/10.1002/eji.200939764
[37]	Latour, S., Roncagalli, R., Chen, R., Bakinowski, M., Shi, X., Schwartzberg, P.L., Davidson, D. and Veillette, A. (2003) Binding of SAP SH2 Domain to FynT SH3 Domain Reveals a Novel Mechanism of Receptor Signalling in Immune Regulation. Nature Cell Biology, 5, 149-154. http://dx.doi.org/10.1038/ncb919
[38]	Qian, D., Lev, S., van Oers, N.S., Dikic, I., Schlessinger, J. and Weiss, A. (1997) Tyrosine Phosphorylation of Pyk2 Is Selectively Regulated by Fyn during TCR Signaling. Journal of Experimental Medicine, 185, 1253-1260. http://dx.doi.org/10.1084/jem.185.7.1253
[39]	Davidson, D., Shi, X., Zhang, S., Wang, H., Nemer, M., Ono, N., Ohno, S., Yanagi, Y. and Veillette, A. (2004) Genetic Evidence Linking SAP, the X-Linked Lymphoproliferative Gene Product, to Src-Related Kinase FynT in TH2 Cytokine Regulation. Immunity, 21, 707-717. http://dx.doi.org/10.1016/j.immuni.2004.10.005
[40]	Beinke, S.R., Phee, H., Clingan, J.M., Schlessinger, J., Matloubian, M. and Weiss, A. (2010) Proline-Rich Tyrosine Kinase-2 Is Critical for CD8 T-Cell Short-Lived Effector Fate. Proceedings of the National Academy of Sciences of the United States of America, 107, 16234-16239. http://dx.doi.org/10.1073/pnas.1011556107
[41]	Veale, M., Raab, M., Li, Z., da Silva, A.J., Kraeft, S.K., Weremowicz, S., Morton, C.C. and Rudd, C.E. (1999) Novel Isoform of Lymphoid Adaptor FYN-T-Binding Protein (FYB-130) Interacts with SLP-76 and Up-Regulates Interleukin 2 Production. Journal of Biological Chemistry, 274, 28427-28435. http://dx.doi.org/10.1074/jbc.274.40.28427
[42]	Griffiths, E.K., Krawczyk, C., Kong, Y.-Y., Raab, M., Hyduk, S.J., Bouchard, D., Chan, V.S., Kozieradzki, I., Oliveira-dos-Santos, A.J., Wakeham, A., et al. (2001) Positive Regulation of T Cell Activation and Integrin Adhesion by the Adapter Fyb/Slap. Science, 293, 2260-2263. http://dx.doi.org/10.1126/science.1063397
[43]	Yablonski, D., Kadlecek, T. and Almeida, A.R. (2001) Identification of a Phospholipase C-γ1 (PLC-γ1) SH3 Domain-Binding Site in SLP-76 Required for T-Cell Receptor-Mediated Activation of PLC-γ1 and NFAT. Molecular and Cellular Biology, 21, 4208-4218. http://dx.doi.org/10.1128/MCB.21.13.4208-4218.2001
[44]	Meldrum, E., Parker, P.J. and Carozzi, A. (1991) The PtdIns-PLC Superfamily and Signal Transduction. Biochimica et Biophysica Acta, 1092, 49-71. http://dx.doi.org/10.1016/0167-4889(91)90177-Y
[45]	Hogan, P.G., Chen, L., Nardone, J. and Rao, A. (2003) Transcriptional Regulation by Calcium, Calcineurin, and NFAT. Genes & Development, 17, 2205-2232. http://dx.doi.org/10.1101/gad.1102703