Laboratory of Tumor Immunology

  • Production and characterization of murine and rabbit monoclonal antibodies recognizing antigenic structures present on the surface of cancer cells
  • Understanding the role of adapter proteins in the ontogenesis of T lymphocytes and the pathogenesis of T cell lymphomas
  • Assessment of the diversity of the TCR repertoire of regulatory and effector T lymphocytes during experimental cancer immunotherapy based on the use of monoclonal antibodies that are inhibitors of immune checkpoints
  • Characterization of a new strain of mice carrying a spontaneous point mutation in the non-erythrocyte spectrin gene as a research model for West’s syndrome
Laboratory Head

Head: prof. dr hab. Arkadiusz Miążek,  (https://orcid.org/0000-0002-7210-2706)

Arkadiusz MiążekArkadiusz Miążek obtained his Ph.D. in biochemistry (1993) from Wroclaw University (Poland) after completing doctoral studies at Basel Institute for Immunology (Switzerland). From 1997 to 2003 he was a postdoctoral researcher at Centre d’Immunologie de Marseille Luminy (France). Since 2011 he holds the position of Head of Laboratory of Tumor Immunology at Hirszfeld Institute of Immunology.

Team

Professors of the Institute:

  • prof. Paweł Kisielow

PhD Students:

  • mgr Michał Zalas
  • mgr inż. Joanna Skrzymowska

Most important scientific achievements
  • Generation and characterization of new cytotoxic monoclonal antibodies against canine lymphomas and leukemias. These are antibodies with therapeutic potential constituting a model for experimental cancer immunotherapy in comparative and veterinary medicine.
  • Characterization of the expression mechanism and function of LAT adapter protein isoforms. A new LAT protein isoform and its diverse expression in the animal world have been described. The genetic elements influencing the formation of this isoform and its function were identified.
  • Establishment of the effect of anti-GITR antibody immunotherapy on remodeling the TCR repertoire of regulatory and effector T cells during the anti-tumor response. It was found that as a result of immunotherapy, there is an increase in the diversity of the TCR repertoire of T-reg lymphocytes infiltrating the tumor and direct competition between effector and regulatory lymphocytes for the same antigens, which may limit the effectiveness of the therapy.

Research methods
  • generation and selection of hybridoma cells producing murine and rabbit monoclonal antibodies
  • antibody engineering: production of chimeric antibodies, production of antibody fragments and their conjugation
  • molecular biology, production of transgenic animals, silencing and overexpression of genes in vitro and in vivo, overproduction of recombinant proteins
  • ELISA, ELISPOT tests, flow cytometry
  • metabolic and behavioral tests in laboratory mouse models

Key equipment
  • table top flow cytometer
  • ELISPOT scanner
  • nanodrop spectrophotometer
  • rotarod: apparatus for studying the motor coordination of mice
  • treadmill for measuring coordination of mouse movements (parallel rod floor test apparatus)
  • blood gas sensors with software to analyze the basal metabolism in mice

Most important projects (during the last 10 years)
  • 2018 – 2021 OPUS 13 Fri. “Characterization of the phenotype of mice carrying a spontaneous, dominant point mutation in the Spna2 gene encoding the alpha II chain of non-erythrocyte spectrin” UMO-2017/25 / B / NZ3 / 03012
  • 2016-2019 PRELUDIUM pt. “Conformational epitopes of the canine DR main histocompatibility complex (DLA-DR) molecules as new targets in the diagnosis and therapy of lymphomas” 2016/21 / N / NZ5 / 01942
  • 2013 – 2015 OPUS 4 pt. “Understanding the suppressive role of the LAT adapter protein in the development of T-type acute lymphoblastic leukemia (T-ALL)” Awarded by the National Science Center (contract no. UMO-2012/05 / B / NZ5 / 01339)
  • 2009 – 2012 own research project of the Ministry of Science and Higher Education N401057237 Study of the expression and localization of the LAT adapter protein among precursors of T lymphocytes in the new LAT-INV-CFP transgenic mouse model

Selected publications
  • An Antibody Specific for the Dog Leukocyte Antigen DR (DLA-DR) and Its Novel Methotrexate Conjugate Inhibit the Growth of Canine B Cell Lymphoma. Lisowska M, Milczarek M, Ciekot J, Kutkowska J, Hildebrand W, Rapak A, Miazek A. Cancers (Basel). 2019 Sep 26;11(10):1438. doi: 10.3390/cancers11101438.
    PMID: 31561563 Free PMC article.
  • Development of novel monoclonal antibodies to dog leukocyte antigen DR displaying direct and immune-mediated cytotoxicity toward canine lymphoma cell lines. Lisowska M, Pawlak A, Kutkowska J, Hildebrand W, Ugorski M, Rapak A, Miazek A. Hematol Oncol. 2018 Mar 24. doi: 10.1002/hon.2507. Online ahead of print.
    PMID: 29573261
  • A Stretch of Negatively Charged Amino Acids of Linker for Activation of T-Cell Adaptor Has a Dual Role in T-Cell Antigen Receptor Intracellular Signaling. Arbulo-Echevarria MM, Narbona-Sánchez I, Fernandez-Ponce CM, Vico-Barranco I, Rueda-Ygueravide MD, Dustin ML, Miazek A, Duran-Ruiz MC, García-Cózar F, Aguado E. Front Immunol. 2018 Feb 2;9:115. doi: 10.3389/fimmu.2018.00115. eCollection 2018.
    PMID: 29456532 Free PMC article.  
  • Anti-GITR Antibody Treatment Increases TCR Repertoire Diversity of Regulatory but not Effector T Cells Engaged in the Immune Response Against B16 Melanoma. Scirka B, Szurek E, Pietrzak M, Rempala G, Kisielow P, Ignatowicz L, Miazek A. Arch Immunol Ther Exp (Warsz). 2017 Dec;65(6):553-564. doi: 10.1007/s00005-017-0479-1. Epub 2017 Jun 21.
    PMID: 28638937 Free PMC article.  
  • Phorbol ester-mediated re-expression of endogenous LAT adapter in J.CaM2 cells: a model for dissecting drivers and blockers of LAT transcription. Marek-Bukowiec K, Aguado E, Miazek A. Genes Immun. 2016 Jul;17(5):313-20. doi: 10.1038/gene.2016.25. Epub 2016 Jun 9.
    PMID: 27278128 Free PMC article.  
  • Identification of functional, short-lived isoform of linker for activation of T cells (LAT). Kłossowicz M, Marek-Bukowiec K, Arbulo-Echevarria MM, Ścirka B, Majkowski M, Sikorski AF, Aguado E, Miazek A. Genes Immun. 2014 Oct;15(7):449-56. doi: 10.1038/gene.2014.35. Epub 2014 Jul 10.
    PMID: 25008862  
  • The membrane adaptor LAT is proteolytically cleaved following Fas engagement in a tyrosine phosphorylation-dependent fashion. García-Blesa A, Klossowicz M, López-Osuna C, Martínez-Florensa M, Malissen B, García-Cózar FJ, Miazek A, Aguado E. Biochem J. 2013 Mar 15;450(3):511-21. doi: 10.1042/BJ20121135.
    PMID: 23240581  
  • Assessment of caspase mediated degradation of linker for activation of T cells (LAT) at a single cell level. Kłossowicz M, Scirka B, Suchanek J, Marek-Bukowiec K, Kisielow P, Aguado E, Miazek A. J Immunol Methods. 2013 Mar 29;389(1-2):9-17. doi: 10.1016/j.jim.2012.12.004. Epub 2012 Dec 20.
    PMID: 23261919  
  • Peripheral Thy1+ lymphocytes rearranging TCR-gammadelta genes in LAT-deficient mice. Miazek A, Macha K, Łaszkiewicz A, Kissenpfennig A, Malissen B, Kisielow P. Eur J Immunol. 2009 Sep;39(9):2596-605. doi: 10.1002/eji.200939252. PMID: 19701892   Restrictase free generation of targeting vectors for disruption of complex mouse genes. Miazek A, Cebula A, Skwarek M, Cebrat M, Kisielow P. Biochem Biophys Res Commun. 2007 Jun 29;358(2):483-8. doi: 10.1016/j.bbrc.2007.04.131. Epub 2007 Apr 30.
    PMID: 17498650