Abstract
In the past 20 years, the interest for the tumor microenvironment (TME) has exponentially increased. Indeed, it is now commonly admitted that the TME plays a crucial role in cancer development, maintenance, immune escape and resistance to therapy. This stands true for hematological malignancies as well. A considerable amount of newly developed therapies are directed against the cancer-supporting TME instead of targeting tumor cells themselves. However, the TME is often not clearly defined. In addition, the unique phenotype of each tumor and the variability among patients limit the success of such therapies. Recently, our group took advantage of the mass cytometry technology to unveil the specific TME in the context of chronic lymphocytic leukemia (CLL) in mice. We found the enrichment of LAG3 and PD1, two immune checkpoints. We tested an antibody-based immunotherapy, targeting these two molecules. This combination of antibodies was successful in the treatment of murine CLL. In this methods article, we provide a detailed protocol for the staining of CLL TME cells aiming at their characterization using mass cytometry. We include panel design and validation, sample preparation and acquisition, machine set-up, quality control, and ...Continue Reading
References
Jun 19, 2010·Nature Reviews. Immunology·Shimon SakaguchiDavid A Hafler
Oct 14, 2010·Blood·Yair HerishanuAdrian Wiestner
Oct 4, 2011·Nature Biotechnology·Peng QiuSylvia K Plevritis
Jun 14, 2012·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Gregory K BehbehaniGarry P Nolan
Aug 21, 2012·Nature Biotechnology·Bernd BodenmillerGarry P Nolan
May 21, 2013·Nature Biotechnology·El-ad David AmirDana Pe'er
Jul 1, 2014·Proceedings of the National Academy of Sciences of the United States of America·Robert V BruggnerGarry P Nolan
Feb 24, 2015·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Sabine TricotAntonio Cosma
Jun 23, 2015·Cell·Jacob H LevineGarry P Nolan
Dec 22, 2015·Methods in Molecular Biology·Albert C Shaw
May 3, 2016·Cell Systems·Bernd Bodenmiller
May 18, 2016·Nature Methods·Nikolay SamusikGarry P Nolan
Sep 16, 2016·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Chikara TakahashiWilliam E O'Gorman
Oct 31, 2016·Experimental Hematology·Rachael J M Bashford-RogersPaul Kellam
Dec 10, 2016·Cytometry. Part B, Clinical Cytometry·Cara Ellen WogslandKanutte Huse
Apr 2, 2017·Physiology & Behavior·P E BunneyC M Kotz
Apr 4, 2017·Current Protocols in Molecular Biology·Nalin LeelatianJonathan M Irish
Apr 27, 2017·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Jamila Elhmouzi-YounesAnne-Sophie Beignon
Nov 25, 2017·Nature Communications·Vincent van UnenBoudewijn P F Lelieveldt
Dec 20, 2017·The Journal of Immunology : Official Journal of the American Association of Immunologists·Abigail K KimballEric T Clambey
Feb 15, 2018·Blood·Marina WierzEtienne Moussay
Apr 2, 2018·Cell Systems·Stéphane ChevrierBernd Bodenmiller
Jun 9, 2018·Blood·Fathima Zumla CaderMargaret A Shipp
Sep 18, 2018·Oncoimmunology·Marina WierzJerome Paggetti
Sep 28, 2018·Nature Protocols·Guojun HanGarry P Nolan
Oct 3, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Lars R OlsenHolden Terry Maecker
Nov 9, 2018·British Journal of Haematology·Daphne R FriedmanJ Brice Weinberg
Nov 20, 2018·Computational and Structural Biotechnology Journal·Guillaume BeyrendRamon Arens
Feb 21, 2019·Cell Reports·Zhi-Zhang YangStephen M Ansell
Sep 11, 2019·Methods in Molecular Biology·Bita SahafSean C Bendall
Oct 22, 2019·European Journal of Immunology·Andrea CossarizzaArturo Zychlinsky
Nov 7, 2019·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Christina Bligaard Pedersen, Lars Rønn Olsen
Feb 13, 2020·Proceedings of the National Academy of Sciences of the United States of America·Palash C MaityHassan Jumaa