An autologous endothelial cell:peripheral blood mononuclear cell assay that detects cytokine storm responses to biologics

FASEB J. 2015 Jun;29(6):2595-602. doi: 10.1096/fj.14-268144. Epub 2015 Mar 6.

Abstract

There is an urgent unmet need for human tissue bioassays to predict cytokine storm responses to biologics. Current bioassays that detect cytokine storm responses in vitro rely on endothelial cells, usually from umbilical veins or cell lines, cocultured with freshly isolated peripheral blood mononuclear cells (PBMCs) from healthy adult volunteers. These assays therefore comprise cells from 2 separate donors and carry the disadvantage of mismatched tissues and lack the advantage of personalized medicine. Current assays also do not fully delineate mild (such as Campath) and severe (such as TGN1412) cytokine storm-inducing drugs. Here, we report a novel bioassay where endothelial cells grown from stem cells in the peripheral blood (blood outgrowth endothelial cells) and PBMCs from the same donor can be used to create an autologous coculture bioassay that responds by releasing a plethora of cytokines to authentic TGN1412 but only modestly to Campath and not to control antibodies such as Herceptin, Avastin, and Arzerra. This assay performed better than the traditional mixed donor assay in terms of cytokine release to TGN1412 and, thus, we suggest provides significant advancement and a definitive system by which biologics can be tested and paves the way for personalized medicine.

Keywords: TGN1412; coculture; personalized medicine; stem cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alemtuzumab
  • Antibodies, Monoclonal / pharmacology
  • Antibodies, Monoclonal, Humanized / pharmacology
  • Bevacizumab
  • Biological Assay / methods
  • Biological Products / pharmacology*
  • Cell Proliferation / drug effects
  • Coculture Techniques
  • Culture Media / pharmacology
  • Cytokines / metabolism*
  • Endothelial Cells / cytology
  • Endothelial Cells / drug effects*
  • Endothelial Cells / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Human Umbilical Vein Endothelial Cells / cytology
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Interleukin-2 / metabolism
  • Interleukin-6 / metabolism
  • Interleukin-8 / metabolism
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / drug effects*
  • Leukocytes, Mononuclear / metabolism
  • Reproducibility of Results
  • Serum / chemistry
  • Trastuzumab
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Humanized
  • Biological Products
  • Culture Media
  • Cytokines
  • Interleukin-2
  • Interleukin-6
  • Interleukin-8
  • Tumor Necrosis Factor-alpha
  • Bevacizumab
  • Alemtuzumab
  • ofatumumab
  • Trastuzumab
  • TGN-1412