EuroMAbNet: European Monoclonal Antibodies Network

Members

University of Oxford

Name of the laboratory

Laboratory activity

Prof Alison Banham’s Haemato-oncology group are focussed on identifying and understanding molecular pathways that are fundamentally required for the pathogenesis of human malignancies (particularly B-cell lymphomas). The aim is to then translate this knowledge into the development of novel cancer biomarkers and therapies. The group are particularly experienced in monoclonal antibody production and characterisation and are actively involved in the European Monoclonal Antibodies Network. Together with its extensive network of collaborators the laboratory uses a multidisciplinary experimental approach exploiting a wide range of biochemical, structural, immunological, pathological, cell biology, molecular, and bioinformatics techniques to pursue its research goals.

Research activities

1) Therapeutic Antibody Programme

Monoclonal antibodies represent some of the most effective new drugs being used to treat cancer. We have produced novel antibodies targeting the tumour vasculature that recognise both known (the Notch ligand Jagged-1) and novel proteins with a role in angiogenesis, with the aim of generating new drugs that disrupt the tumour blood supply. We have also generated TCR mimic antibodies to wild type p53 peptides in the context of HLA-A2. These extend the scope of classical therapeutic antibody targets to include intracellular tumour proteins via the specific recognition of tumour-derived peptides presented on the tumour cell surface by MHC class I molecules.

2) Transcriptional deregulation in cancer

The altered patterns of gene expression arising from inappropriate regulation and/or function of key transcription factors cause many human diseases, including cancer. We previously discovered the human FOXP1 forkhead transcription factor and it’s roles in the pathogenesis of both lymphomas and carcinomas, while we identified overexpression of the related FOXP2 protein in the incurable malignancy multiple myeloma. In collaboration with Dr Giovanna Roncador, we were the first to make monoclonal antibodies to the regulatory T-cell marker FOXP3, and have demonstrated across a number of tumour types that these reagents can be used to quantify the numbers of these cells in patients’ biopsies and predict clinical outcomes such as survival or late relapse. Further studies of the FOXP family, NFIL3 (a transcription factor essential for the NK lineage) and the PASD1 cancer testis antigen are being pursued by studying their expression, regulation, involvement in pathways that affect cellular survival and proliferation, combined with the identification and characterisation of their co-factors and target genes. Future work will include hit identification and drug discovery studies to identify FOXP inhibitors.

Techniques available

• monoclonal antibody production in mice
• gene cloning and preparation of transfection constructs
• recombinant protein expression in bacteria and mammalian cells
• antibody characterisation
• RNAi to confirm antibody specificity
• single, double and triple immunostaining techniques
• bacterial expression cloning of antigens recognised by antibodies
• immunological techniques eg. immunoprecipitation and western blotting
• Chromatin immunoprecipitation
• Characterisation of humoral immune responses to tumour proteins
• Antibody purification
• Antibody gene cloning, recombinant antibody expression, isotype switching and production of chimeric antibodies
• Production and characterisation of TCR mimic antibodies
• Testing candidate therapeutic antibodies in pre-clinical xenograft models
• CRISPR/Cas9 genome engineering

Publications (2019-present)

• Boullosa LF, Bonney S, Savaliya P, Orchard L, Wickenden H, Lee C, Smits E, Banham AH, Mills K, Orchard K, Guinn B-A. Identification of survivin as the most promising leukaemia associated antigen for the immunotherapy of adult B-cell acute lymphoblastic leukaemia. Oncotarget 2017 Dec 17;9(3):3853-3866. doi: 10.18632/oncotarget.23380. eCollection 2018 Jan 9.
• Ait-Tahar K, Anderson AP, Barnardo M, Collins GP, Hatton C, Banham AH, Pulford K. Sp17 protein expression and major histocompatibility class I and II epitope presentation in diffuse large B-cell lymphoma patients. Advances in Hematology 2017;2017:6527306. doi: 10.1155/2017/6527306.
• Loo SK, Ch’ng ES, Lawrie CH, Arestin Muruzabal M, Gaafar A, Puente Pomposo M, Husin A, Salleh S, Banham AH, Pedersen LM, Møller MB, Green TM, Wong KK. DNMT1 is predictive of survival and associated with Ki-67 expression in R-CHOP-treated diffuse large B-cell lymphomas. Pathology 2017 49(7):731-739. doi: 10.1016/j.pathol.2017.08.009.
• Trenevska I, Li D, Banham AH. Therapeutic antibodies against intracellular tumor antigens. Frontiers in Immunology 2017 Aug;8: article 1001. doi: 10.3389/fimmu.2017.01001
• Li D, Bentley C, Yates J, Salimi M, Greig J, Wiblin S, Hassanali T, Banham AH. Engineering chimeric human and mouse major histocompatibility (MHC) class I tetramers for the production of T-cell receptor (TCR) mimic antibodies. PLOS One 2017 12(4):e0176642.
• Li D, Bentley C, Anderson A, Wiblin S, Cleary KLS, Koustoulidou S, Hassanali T, Yates J, Greig J, Olde Nordkamp M, Trenevska I, Ternette N, Kessler BM, Cornelissen B, Cragg MS, Banham AH. Development of a T-cell receptor mimic antibody against wild type p53 for cancer immunotherapy. Cancer Research 2017;77(10):2699-711.
• Loo SK, Ch’ng ES, Salleh S, Banham AH, Pedersen LM, Møller MB, Green TM, Wong KK. TRPM4 expression is associated with activated B-cell subtype and poor survival in diffuse large B-cell lymphoma. Histopathology. 2017 Mar 1. doi: 10.1111/his.13204. [Epub ahead of print]
• Gascoyne DM, Lyne L, Spearman H, Buffa F, Soilleux EJ, Banham AH. Vitamin D receptor expression in plasmablastic lymphoma and myeloma cells confer susceptibility to vitamin D. Endocrinology. 2017 Mar;158(3):503-515.
• G. Roncador, P. Engel, L. Maestre, A.P. Anderson, J.L. Cordell, M.S. Cragg, V.C. Šerbec. M. Jones, V.L. Lisnic, L. Kremer, D. Li, F. Koch-Nolte, N. Pascual, J.I. Rodríguez-Barbosa, R. Torensma, H. Turley, K. Pulford, A.H. Banham, 2016. The European Antibody Network’s practical guide to finding and validating suitable antibodies for research. Mabs 8(1):27-36.
• Gascoyne DM, Banham AH. The significance of FOXP1 in diffuse large B-cell lymphoma. Leuk Lymphoma. 2017 May;58(5):1037-1051. doi: 10.1080/10428194.2016.1228932. Epub 2016 Sep 27.
• Wong KK, Gascoyne DM, Soilleux EJ, Lyne L, Spearman H, Roncador G, Pedersen LM, Møller MB, Green TM, Banham AH. FOXP2-positive diffuse large B-cell lymphomas exhibit a poor response to R-CHOP therapy and distinct biological signatures. Oncotarget. 2016 Aug 16;7(33):52940-52956. doi: 10.18632/oncotarget.9507.
• Brown PJ*, Gascoyne DM*, Lyne L, Spearman H, Felce SL, Chakravarty P, Barrans S, Lynham S, Calado DP, Ward M, Banham AH. N-terminally truncated FOXP1 protein expression and alternate internal FOXP1 promoter usage in normal and malignant B cells. Haematologica. 2016 Jul;101(7):861-71. doi: 10.3324/haematol.2016.142141. Epub 2016 Apr 7.
• Gu Y, Masiero M and Banham AH. Notch signaling: its roles and therapeutic potential in hematological malignancy. Oncotarget. 2016 May 17;7(20):29804-23. doi: 10.18632/oncotarget.7772.
• Brown PJ, Wong KK, Felce SL, Lyne L, Spearman H, Soilleux EJ, Pederson LM, Moller MB, Green TM, Gascoyne DM, Banham AH. FOXP1 suppresses immune response signatures and MHC class II expression in activated B-cell-like diffuse large B-cell lymphomas. Leukemia. 2016 Mar;30(3):605-16. doi: 10.1038/leu.2015.299. Epub 2015 Oct 26.
• S.E. Brooks, S.A. Bonney, C. Lee, A. Publicover, G. Khan, E.L. Smits, D. Sigurdardottir, M. Arno, D. Li, K.L. Mills, K. Pulford, A.H. Banham, V. van Tendeloo, G.J. Mufti, H-G Rammensee, T.J. Elliot, K.H. Orchard, B-A Guinn, 2015. Application of the pMHC array to characterise tumour antigen specific T cell populations in leukaemia patients at disease diagnosis. PLOS One Oct 22;10(10):e0140483. doi: 10.1371/journal.pone.0140483. eCollection 2015
• K.K. Wong, E.S. Ch’ng, S.K. Loo, A. Husin, M.A. Muruzabal, M.B. Møller, L.M. Pedersen, M.P. Pomposo, A. Gaafar, A.H. Banham, T.M. Green, C.H. Lawrie, 2015. Low HIP1R mRNA and protein expression are associated with worse survival in diffuse large B-cell lymphoma patients treated with R-CHOP. Expt Mol Path 99: 537-545.
• G. Deng, Y. Nagai, Y. Xiao, Z. Li, S. Dai, T. Ohtani, A. Banham, B. Li, H. Zhang, S-L. Wu, W. Hancock, A. Samanta, H. Zhang, M.I. Greene, 2015. Pim-2 kinase influences T regulatory cell function and stability by mediating Foxp3 N-terminal phosphorylation. J Biol Chem 290(33):20211-20.
• D.M. Gascoyne, H. Spearman, L. Lyne, R. Puliyadi, M. Pérez-Alcántara, L. Coulton, S.E. Fisher, P.I. Croucher, A. H. Banham, 2015. The forkhead transcription factor FOXP2 is required for regulation of p21WAF1/CIP1 in 143B osteosarcoma cell growth arrest. PLOS One 10(6):e0128513.
• A.K. Michael, S.L. Harvey, P.J. Sammons, A.P. Anderson, H.M. Kopalle, A.H. Banham, C.L. Partch, 2015. Cancer/testis antigen PASD1 silences the circadian clock. Molecular Cell 58:743-54.
• J.F. Collin, J.W. Wells, B. Czepulkowski, L. Lyne, P.J. Duriez, A.H. Banham, G.J. Mufti, B-A Guinn. A novel zinc finger gene, ZNF465, is inappropriately expressed in acute myeloid leukaemia cells. Genes Chromosomes and Cancer 54:288-302.
• D.M. Favara, A.H. Banham, A.L. Harris, 2014. A review of ELTD1, a pro-angiogenic adhesion GPCR. Biochem Soc Trans. 42(6):1658-64.
• D. Li, M. Masiero, A.H. Banham, A.L. Harris, 2014. The Notch ligand Jagged1 as a target for anti-tumour therapy. Front Oncol. Sep 25;4:254. Doi 10.3389/fonc.2014.00254
• T. Negoro, S. Shimizu, M. Narushima, A.H. Banham, H. Wakabayashi, R. Takayanagi, T. Hagiwara, G. Roncador, T. Osabe, T. Yanai, M. Kin, K. Ikeda, A. Endo, H. Akiyama, Y. Nakano, 2014. Elevated receptor for activated C kinase 1 expression is involved in intra-cellular Ca2+ influx and potentially associated with compromised Tregs function in patients with asthma. Clin. Expt. Allergy. 44(9):1154-69.
• S. Patel, D. Murphy, E. Haralambieva, Z.A. Abdulla, K-K. Wong, H. Chen, E. Gould, G. Roncador, C.S.R. Hatton, A.P. Anderson, A.H. Banham, K. Pulford, 2014. Increased expression of phosphorylated FADD in anaplastic large cell and other T-cell lymphomas. Biomarker Insights 9: 77-84.
• K.-K. Wong, D.M. Gascoyne, P.J. Brown, E. Soilleux, C. Snell, H. Chen, L. Lyne, C.H. Lawrie, R.D. Gascoyne, L. M. Pedersen, M.B. Møller, K. Pulford, D. Murphy, T. M. Green, A.H. Banham, 2014. Reciprocal expression of the endocytic protein HIP1R and its repressor FOXP1 predicts outcome in R-CHOP-treated diffuse large B-cell lymphoma patients. Leukemia 28(2):362-72.
• S.K. Williams, E. Donaldson, T. Van der Kleij, L. Dixon, M. Fisher, J. Tibble, Y. Gilleece, P. Klenerman, A.H. Banham, M. Howard, D.P. Webster. Quantification of hepatic FOXP3+ T-lymphocytes in HIV-hepatitis C co-infection. J Viral Hepat 21(4):251-9.
• P. Whiteman, B. Hernandez de Madrid, P. Tayor, D. Li, R. Heslop, N. Viticheep, J. Zi Tan, H. Shimizu, J. Callaghan, M. Masiero, J-L. Li, A.H. Banham, A.L. Harris, S.M. Lea, C. Redfield, M. Baron, P.A. Handford. 2013. Molecular basis for Jagged-1/Serrate recognition by Notch. J Biol Chem. 288:7305-12.
• S. Patel, H. Chen, L. Monti, E. Gould, E. Haralambieva, J. Schmid, D. Toomey, W. Woessmann, G. Roncador, C.S.R. Hatton, A.P. Liggins, R. Taramelli, A.H. Banham, F. Acquati, D. Murphy, K. Pulford. 2012. RNASET2 – an autoantigen in anaplastic large cell lymphoma identified by protein array analysis. J Proteomics 75:5279-92.
• D Yusuf, S.L Butland, M.I Swanson, E Bolotin, A Ticoll, W.A Cheung, X Yu, C Zhang, C.T.D Dickman, D.L Fulton, J.S Lim, J.M Schnabl, O.H.P Ramos, M Vasseur- Cognet, C.N de Leeuw, E.M Simpson, G.U Ryffel, E.W-F Lam, R Kist, M.S.C Wilson, R Marco-Ferreres, J.J Brosens, L.L Beccari, P Bovolenta, B.A Benayoun, L.J Monteiro, H.D.C Schwenen, L Grontved, E Wederell, S Mandrup, R.A Veitia, H Chakravarthy, P.A Hoodless, M.M Mancarelli, B.E Torbett, A.H Banham, S.P Reddy, R.L Cullum, M Liedtke, M.P Tschan, M Vaz, A Rizzino, M Zannini, S Frietze, P.J Farnham, A Eijkelenboom, P.J Brown, D Laperrière, D Leprince, T de Cristofaro, K.L Prince, M Putker, L del Peso, G Camenisch, R.H Wenger, M Mikula, M Rozendaal, S Mader, J Ostrowski, S.J Rhodes, C Van Rechem, G Boulay, S.W.Z Olechnowicz, M.B Breslin, M.S Lan, K.K Nanan, M Wegner, J Hou, R.D Mullen, S.C Colvin, P.J Noy, C.F Webb, M.E Witek, S Ferrell, J.M Daniel, J Park, S.A Waldman, D.J Peet, M Taggart, P-S Jayaraman, J.J Karrich, B Blom, F Vesuna, H O'Geen, Y Sun, R.M Gronostajski, M.W Woodcroft, M.R Hough, E Chen, G.N Europe-Finner, M Karolczak-Bayatti, J Bailey, O Hankinson, V Raman, D.P LeBrun, S Biswal, C.J Harvey, J.P DeBruyne, J.B Hogenesch, R.F Hevner, C Héligon, X.M Luo, M.C Blank, K.J Millen, D.S Sharlin, D Forrest, K Dahlman- Wright, C Zhao, Y Mishima, S Sinha, R Chakrabarti, E Portales-Casamar, F.M Sladek, P.H Bradley and W.W Wasserman. The Transcription Factor Encyclopedia. Genome Biology 13:R24 (2012).
• T. Negoro, S. Shimizu, Y. Yamamoto, A.H. Banham, G. Roncador, H. Akiyama, Y. Nakano. 2012. Chapter 4 - Immune Mechanisms of Childhood Asthma p77-88, in ‘Bronchial Asthma’ publisher InTec Open Access.
• C.D.O. Cooper, C.H. Lawrie, A.P. Liggins, G.P. Collins, C.S. Hatton, K. Pulford & A.H. Banham (2011). Serological identification of antigens associated with peripheral T-cell lymphoma (PTCL). PLOS One. 6(8):e23916.
• M. Yan, N. Jene, D. Byrne, E.K.A. Millar, S.A. O’Toole, C.M. McNeil, G.J. Bates, A.L. Harris, A.H. Banham, R.L. Sutherland, S.B. Fox (2011). Recruitment of regulatory T cells is driven by hypoxia induced CXCR4 expression, and is associated with poor prognosis in basal-like breast cancers. Breast Cancer Res 13(2):R47.
• Y. Yamamoto, T. Negoro, A. Hoshi, A. Wakagi, S. Shimizu, A.H. Banham, M. Ishii, H. Akiyama, Y. Kiuchi, S. Sunaga, T. Tobe, G. Roncador, K. Itabashi, Y. Nakano (2011). Impaired Ca2+ regulation of CD4+CD25+ regulatory T cells from pediatric asthma. Int Arch Allergy Immunol. 156(2):148-158.
• K. Ait-Tahar, A.P. Liggins, G.P. Collins, A. Campbell, M. Barnardo, C. Lawrie, D. Moir, C. Hatton, A.H. Banham, K. Pulford, (2011). CD4-positive Th responses to the PASD1 protein in patients with diffuse large B-cell lymphoma. Haematologica 96(1):78-86.
• Y. Yamamoto, T. Negoro, A. Wakagi, A. Hoshi, A.H. Banham, G. Roncador, T. Tobe, S. Sunaga, Y. Nakano, K. Itabashi. (2010). Participation of Th17 and Tregs cells in pediatric bronchial asthma and food allergies. J Health Sci. 56(5):589-97.
• D. Joseph-Pietras, Y. Gao, N. Zojer, K. Ait-Tahar, A.H. Banham, K. Pulford, J. Rice, N. Savelyeva, S.S. Sahota, (2010). DNA vaccines to target the cancer testis antigen PASD1 in human multiple myeloma. Leukemia 24(11):1951-9.
• A.P. Liggins, S.H. Lim, E.J. Soilleux, K. Pulford and A.H. Banham. (2010). A panel of cancer-testis gene exhibiting broad spectrum expression in haematological malignancies. Cancer Immun 10:8.
• A. Chadburn, E. Hyjek, A.H. Banham, K. Bhatia, E. Cesarman (2010). Reply to K. Dunleavy et al. J Clin. Oncol. 28(16):e261-2.
• E. Ballabio, J. Chi, G. Roncador, A.H. Banham, C.S.R. Hatton, C.H. Lawrie(2010). Comparison of Choi and Hans’ algorithms by immunohistochemistry and quantitative reverse transcriptase-PCR. Clin. Cancer Res. 16(14):3805-6.
• L. Kemény, Zs. Csoma, E. Bagdi, A.H. Banham, L. Krenács, A. Koreck (2010). Targeted phototherapy of plaque type psoriasis using UVB-LEDs. Br J Dermatology 163(1):167-73.
• A.J. Campbell, L. Lyne, P.J. Brown, R.J. Launchbury, P.A. Bignone, J. Chi, G. Roncador, C.H. Lawrie, K. Gatter, R. Kusec and A.H. Banham (2010). Aberrant expression of the neuronal transcription factor FOXP2 in neoplastic plasma cells. Br J Haematol. 149(2): 221-30.
• A.H. Banham, L. Lyne, T.L. Scase, B.A. Blacklaws. Monoclonal antibodies raised to the human FOXP3 protein can be used effectively for detecting Foxp3+ T cells in other mammalian species. Journal of Veterinary Immunology and Immunopathology. Epub ahead of print.
• P.A. Bignone & A.H. Banham. FOXP3+ regulatory T cells as biomarkers in human malignancies. Commissioned review for Expert Opin Biol Ther 8:1897-1920.
• P.J. Brown, S.L. Ashe, E. Leich, C. Burek, S. Barrans, J. Fenton, A.S. Jack, K Pulford, A. Rosenwald and A.H Banham, 2008. Potentially oncogenic B-cell activation induced smaller isoforms of FOXP1 are highly expressed in the activated B-cell-like subtype of DLBCL. Blood 111: 2816-2824.
• P.J. Brown, R. Kagaya and A.H. Banham, 2008. Characterisation of human FOXP1 isoform 2, using monoclonal antibody 4E3-G11, and intron retention as a tissue-specific mechanism generating a novel FOXP1 isoform. Histopathol. Epub ahead of print.