- Czech RepublicVáclav HorejsíInstitute of Molecular Genetics AS CR
- FranceAndré PèlegrinInstitut de Recherche en Cancérologie de Montpellier, IRCM
- Armand BensussanHôpital Saint Louis
- ItalyFabio MalavasiUniversity of Torino Medical School
- NetherlandsRuurd TorensmaNijmegen Centre for Molecular Life Sciences
- SpainGiovanna RoncadorCentro Nacional de Investigaciones Oncológicas
- Leonor KremerCentro Nacional de Biotecnología/CSIC
- Núria PascualNb4D
- Pablo EngelMedical School, University of Barcelona
- United KingdomUniversity of OxfordLRF Haemato-oncology Group
- University of OxfordOxFabs
- University of OxfordLRF Lymphoma Antigens Group
University of Oxford
Name of the laboratory
- LRF Haemato-oncology Group
- Nuffield Department of Clinical Laboratory Sciences, University of Oxford
- Level 4 Academic Block
- University of Oxford
- Headington, Oxford, OX3 9DU, United Kingdom
- +44 1865 220246
Our group is supported by a Programme grant from the Leukaemia Research Fund to study aberrant transcription factors in lymphomas. From April 2009 we have been awarded a Therapeutic Antibody Programme from Cancer Research-UK. We aim to make antibodies that will enable us to i) identify novel diagnostic/prognostic markers, ii) study aberrant protein function in human malignancies, iii) provide additional therapeutic antibodies for the treatment of cancer.
FOXP transcription factors: A complex system of tightly regulated gene expression is essential for an organism’s development and survival. The forkhead or winged helix family of proteins, named FOX (forkhead box) transcription factors, display remarkable functional diversity and are involved in many biological processes, for example, development, oncogenesis, signal transduction, cell cycle regulation, control of metabolism and chromatin remodelling.
There are four members of the FOXP subfamily. We originally cloned the human FOXP1 gene and showed that FOXP1 expression is significantly associated with patients’ survival in both breast cancer and diffuse large B-cell lymphoma (DLBCL). We have recently identified smaller B-cell activation induced FOXP1 isoforms that may have an oncogenic role in DLBCL. This is consistent with FOXP1 chromosome translocations in lymphoma commonly targeting the coding region. FOXP3 is a marker of regulatory T cells that mediate immunotolerance and an important biomarker in both carcinomas and lymphoma. We are using specific “in house” monoclonal antibodies to further study the expression and function of the FOXP proteins and are using a wide range of techniques including chromatin immunoprecipitation to identify FOXP target genes, RNA interference for gene silencing and microRNA modulation to identify FOXP regulators.
Lymphoma-associated antigens: We have had a long-standing collaboration with Dr. Karen Pulford (Oxford) to identify and characterise lymphoma-associated proteins that are recognised by circulating antibodies in patients’ sera. We originally used the SEREX technique to identify a wide variety of lymphoma-associated antigens in diffuse large B-cell lymphoma (DLBCL). This approach identified PAS domain containing 1 (PASD1), a novel transcription factor, as a cancer testis antigen and candidate for vaccine development for lymphoma immunotherapy. The MORC4 zinc finger protein is also of interest because its immunological recognition may reflect its overexpression in a subgroup of DLBCL patients.
A more recent collaboration with Dr. Derek Murphy (Dublin) and Karen Pulford has extended the identification of lymphoma-associated proteins by screening other subtypes of lymphoma using high-throughput protein microarray technology. This has identified yet more exciting molecules for further study, which will contribute to Dublin’s development of array chips for cancer serum screening and Dr. Pulford’s vaccine Programme.
Our functional characterisation of these antigens is critical to understand their role in lymphomagenesis and whether targeting them will inhibit tumour growth and survival. While cancer testis antigens are widely accepted to be valuable targets for therapy, there is still little understanding of their biological role in malignancy.
- 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
- 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.
- C.D.O. Cooper, A.P. Liggins, K. Ait-Tahar, G. Roncador, A.H. Banham, K. Pulford, 2006. Protein expression profiles confirm PASD1 as a cancer-testis antigen and a potential candidate for lymphoma immunotherapy. Leukemia 20, 2172-2174.
- J. Carreras, A. Lopez-Guillermo, B.C. Fox, L. Colomo, A. Martinez, G. Roncador, E. Monserrat, E. Campo and A.H. Banham, 2006. High numbers of tumor infiltrating FOXP3-positive regulatory T cells are associated with improved overall survival in follicular lymphoma. Blood 108, 2957-2964.
- G. Roncador, P.J. Brown, L. Mastre, S. Hue, J.L. Martínez-Torrecuadrada, K-L. Ling, S. Pratap, C. Toms, B.C. Fox, V. Cerundolo, F. Powrie and A.H. Banham, 2005. Analysis of FOXP3 protein expression in human CD4+CD25+ regulatory T cells at the single cell level. Eur. J. Immunol. 35, 1681-1691.
- A.H. Banham, J.M. Connors, P.J. Brown, J.L. Cordell, G. Ott, G. Sreenivasan, P. Farinha, D.E. Horsman and R. Gascoyne, 2005. Expression of the FOXP1 transcription factor is strongly associated with inferior survival in patients with diffuse large B-cell lymphoma. Clin. Cancer Res. 11; 1065-1072.
- A.P. Liggins, G. Guinn, C.S. Hatton, K. Pulford & A.H. Banham, 2004. Serologic detection of diffuse large B-cell lymphoma-associated antigens. International Journal of Cancer 110, 563-569.
- A.H. Banham, N. Beasley, E. Campo, P.L. Fernandez, C. Fidler, K. Gatter, M. Jones, D.Y. Mason, J.E. Prime, P.Trougouboff, K.Wood & J.L. Cordell, 2001. The FOXP1 winged helix transcription factor is a novel candidate tumour suppressor gene on chromosome 3p. Cancer Research, 61, 8820-8829.