Serving the Society Since 1986
Advanced Search
Volume 34 Issue 4
August 2019
    Citation: Gaoxin Li, Ling Ding, Xiaojing Ma, Qiliang Cai, Tianlei Ying, Fang Wei. Establishment of Novel Monoclonal Fabs Specific for Epstein-Barr Virus Encoded Latent Membrane Protein 1 .VIROLOGICA SINICA, 2019, 34(4) : 467-470.  http://dx.doi.org/10.1007/s12250-019-00103-6
    shu

    Establishment of Novel Monoclonal Fabs Specific for Epstein-Barr Virus Encoded Latent Membrane Protein 1

    cstr: 32224.14.s12250-019-00103-6
    • 1.

      Sheng Yushou Center of Cell Biology and Immunology, School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China

    • 2.

      MOE & MOH Key Laboratory of Medical Molecular Virology, School of Basic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China

    • Corresponding author: Fang Wei, fangwei@sjtu.edu.cn, ORCID: 0000-0001-8964-1645
    • Received Date: 22 November 2018
      Accepted Date: 23 January 2019
      Published Date: 04 April 2019
      Available online: 01 August 2019
    • Here we isolated the Fab clones 1-A11, 6-C6, 10-B2, and 15-H10 and confirmed their recognition of LMP1 expressed on the surface of EBV+ B95.8 cells. Recognition of LMP1 on the cell membrane by clone 15-H10 was verified through immunofluorescent assay, while other clones failed the recognition. One of the possible reasons could be the stringent washing in this assay that ruptured the low affinity Fab-LMP1 binding. EBV infection is related with multiple diseases, and conventional approaches used in laboratory diagnostic tests for EBV infection and pathogenesis have their limitations [(generate false positive results or lack the ability to localize the expression of EBV within target cells) (Young and Rickinson 2004)]. The LMP1-specific Fab clones reported here need further evaluation in both affinity and specificity in testing EBV+ patient samples, and hopefully it could have potential applications in EBV diagnostics and directly targeting EBV-related tumors in adoptive T cell therapy.
    • 加载中

      1. Chen R, Zhang D, Mao Y, Zhu J, Ming H, Wen J, Ma J, Cao Q, Lin H, Tang Q, Liang J, Feng Z (2012) A human Fab-based immunoconjugate specific for the LMP1 extracellular domain inhibits nasopharyngeal carcinoma growth in vitro and in vivo. Mol Cancer Ther 11:594-603
          doi: 10.1158/1535-7163.MCT-11-0725

      2. Cho HI, Kim UH, Shin AR, Won JN, Lee HJ, Sohn HJ, Kim TG (2018) A novel Epstein-Barr virus-latent membrane protein-1-specific T-cell receptor for TCR gene therapy. Br J Cancer 118:534-545
          doi: 10.1038/bjc.2017.475

      3. Deacon EM, Pallesen G, Niedobitek G, Crocker J, Brooks L, Rickinson AB, Young LS (1993) Epstein-Barr virus and Hodgkin's disease: transcriptional analysis of virus latency in the malignant cells. J Exp Med 177:339-349
          doi: 10.1084/jem.177.2.339

      4. Dirmeier U, Neuhierl B, Kilger E, Reisbach G, Sandberg ML, Hammerschmidt W (2003) Latent membrane protein 1 is critical for efficient growth transformation of human B cells by Epstein-Barr virus. Cancer Res 63:2982-2989
          doi: 10.1097/00002820-200306000-00012

      5. Fennewald S, van Santen V, Kieff E (1984) Nucleotide sequence of an mRNA transcribed in latent growth-transforming virus infection indicates that it may encode a membrane protein. J Virol 51:411-419

      6. Gires O, Zimber-Strobl U, Gonnella R, Ueffing M, Marschall G, Zeidler R, Pich D, Hammerschmidt W (1997) Latent membrane protein 1 of Epstein-Barr virus mimics a constitutively active receptor molecule. EMBO J 16:6131-6140
          doi: 10.1093/emboj/16.20.6131

      7. Hislop AD, Taylor GS, Sauce D, Rickinson AB (2007) Cellular responses to viral infection in humans: lessons from Epstein-Barr virus. Annu Rev Immunol 25:587-617
          doi: 10.1146/annurev.immunol.25.022106.141553

      8. Kaye KM, Devergne O, Harada JN, Izumi KM, Yalamanchili R, Kieff E, Mosialos G (1996) Tumor necrosis factor receptor associated factor 2 is a mediator of NF-kappa B activation by latent infection membrane protein 1, the Epstein-Barr virus transforming protein. Proc Natl Acad Sci U S A 93:11085-11090
          doi: 10.1073/pnas.93.20.11085

      9. Kutz H, Reisbach G, Schultheiss U, Kieser A (2008) The c-Jun N-terminal kinase pathway is critical for cell transformation by the latent membrane protein 1 of Epstein-Barr virus. Virology 371:246-256
          doi: 10.1016/j.virol.2007.09.044

      10. Lin X, Gudgeon NH, Hui EP, Jia H, Qun X, Taylor GS, Barnardo MC, Lin CK, Rickinson AB, Chan AT (2008) CD4 and CD8 T cell responses to tumour-associated Epstein-Barr virus antigens in nasopharyngeal carcinoma patients. Cancer Immunol Immunother 57:963-975
          doi: 10.1007/s00262-007-0427-8

      11. Lu ZX, Ma XQ, Yang LF, Wang ZL, Zeng L, Li ZJ, Li XN, Tang M, Yi W, Gong JP, Sun LQ, Cao Y (2008) DNAzymes targeted to EBV-encoded latent membrane protein-1 induce apoptosis and enhance radiosensitivity in nasopharyngeal carcinoma. Cancer Lett 265:226-238
          doi: 10.1016/j.canlet.2008.02.019

      12. Omar N, Lim TS (2018) Construction of naive and immune human Fab phage-display library. Methods Mol Biol 1701:25-44
          doi: 10.1007/978-1-4939-7447-4

      13. Pope JH, Horne MK, Scott W (1968) Transformation of foetal human keukocytes in vitro by filtrates of a human leukaemic cell line containing herpes-like virus. Int J Cancer 3:857-866
          doi: 10.1002/(ISSN)1097-0215

      14. Williams H, Crawford DH (2006) Epstein-Barr virus: the impact of scientific advances on clinical practice. Blood 107:862-869

      15. Xiao X, Ho M, Zhu Z, Pastan I, Dimitrov DS (2009) Identification and characterization of fully human anti-CD22 monoclonal antibodies. MAbs 1:297-303
          doi: 10.4161/mabs.1.3.8113

      16. Ying T, Du L, Ju TW, Prabakaran P, Lau CC, Lu L, Liu Q, Wang L, Feng Y, Wang Y, Zheng BJ, Yuen KY, Jiang S, Dimitrov DS (2014) Exceptionally potent neutralization of Middle East respiratory syndrome coronavirus by human monoclonal antibodies. J Virol 88:7796-7805
          doi: 10.1128/JVI.00912-14

      17. Young LS, Rickinson AB (2004) Epstein-Barr virus: 40 years on. Nat Rev Cancer 4:757-768
          doi: 10.1038/nrc1452

    • 加载中

    Figures(1)

    PDF

    Article Metrics

    Article views(8296) PDF downloads(49) Cited by()

    Related
    Proportional views

      Establishment of Novel Monoclonal Fabs Specific for Epstein-Barr Virus Encoded Latent Membrane Protein 1

        Corresponding author: Fang Wei, fangwei@sjtu.edu.cn
      • 1. Sheng Yushou Center of Cell Biology and Immunology, School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China
      • 2. MOE & MOH Key Laboratory of Medical Molecular Virology, School of Basic Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, China
      • Received Date: 22 November 2018
      • Accepted Date: 23 January 2019
      • Published Date: 04 April 2019

      Abstract: Here we isolated the Fab clones 1-A11, 6-C6, 10-B2, and 15-H10 and confirmed their recognition of LMP1 expressed on the surface of EBV+ B95.8 cells. Recognition of LMP1 on the cell membrane by clone 15-H10 was verified through immunofluorescent assay, while other clones failed the recognition. One of the possible reasons could be the stringent washing in this assay that ruptured the low affinity Fab-LMP1 binding. EBV infection is related with multiple diseases, and conventional approaches used in laboratory diagnostic tests for EBV infection and pathogenesis have their limitations [(generate false positive results or lack the ability to localize the expression of EBV within target cells) (Young and Rickinson 2004)]. The LMP1-specific Fab clones reported here need further evaluation in both affinity and specificity in testing EBV+ patient samples, and hopefully it could have potential applications in EBV diagnostics and directly targeting EBV-related tumors in adoptive T cell therapy.

        HTML