Super-resolution microscopy reveals ultra-low CD19 expression on myeloma cells that triggers elimination by CD19 CAR-T

Nat Commun. 2019 Jul 17;10(1):3137. doi: 10.1038/s41467-019-10948-w.

Abstract

Immunotherapy with chimeric antigen receptor-engineered T-cells (CAR-T) is under investigation in multiple myeloma. There are reports of myeloma remission after CD19 CAR-T therapy, although CD19 is hardly detectable on myeloma cells by flow cytometry (FC). We apply single molecule-sensitive direct stochastic optical reconstruction microscopy (dSTORM), and demonstrate CD19 expression on a fraction of myeloma cells (10.3-80%) in 10 out of 14 patients (density: 13-5,000 molecules per cell). In contrast, FC detects CD19 in only 2 of these 10 patients, on a smaller fraction of cells. Treatment with CD19 CAR-T in vitro results in elimination of CD19-positive myeloma cells, including those with <100 CD19 molecules per cell. Similar data are obtained by dSTORM analyses of CD20 expression on myeloma cells and CD20 CAR-T. These data establish a sensitivity threshold for CAR-T and illustrate how super-resolution microscopy can guide patient selection in immunotherapy to exploit ultra-low density antigens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD19 / metabolism*
  • Biomarkers / metabolism
  • Cohort Studies
  • Flow Cytometry
  • Humans
  • Immunotherapy, Adoptive
  • Interferon-gamma / metabolism
  • Multiple Myeloma / metabolism
  • Multiple Myeloma / therapy*
  • Receptors, Chimeric Antigen / therapeutic use

Substances

  • Antigens, CD19
  • Biomarkers
  • CD19 molecule, human
  • Receptors, Chimeric Antigen
  • Interferon-gamma