Clinical microfluidics for neutrophil genomics and proteomics

Nat Med. 2010 Sep;16(9):1042-7. doi: 10.1038/nm.2205. Epub 2010 Aug 29.

Abstract

Neutrophils have key roles in modulating the immune response. We present a robust methodology for rapidly isolating neutrophils directly from whole blood with 'on-chip' processing for mRNA and protein isolation for genomics and proteomics. We validate this device with an ex vivo stimulation experiment and by comparison with standard bulk isolation methodologies. Last, we implement this tool as part of a near-patient blood processing system within a multi-center clinical study of the immune response to severe trauma and burn injury. The preliminary results from a small cohort of subjects in our study and healthy controls show a unique time-dependent gene expression pattern clearly demonstrating the ability of this tool to discriminate temporal transcriptional events of neutrophils within a clinical setting.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibodies, Monoclonal
  • Antigens, CD / genetics
  • Antigens, CD / immunology
  • Biotinylation
  • Burns / physiopathology*
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / immunology
  • DNA / genetics
  • DNA / isolation & purification
  • GPI-Linked Proteins
  • Genomics / methods*
  • Humans
  • Microfluidics / methods*
  • Neutrophils / cytology
  • Neutrophils / physiology*
  • Oligonucleotide Array Sequence Analysis
  • Proteomics / methods*
  • RNA / genetics
  • RNA / isolation & purification
  • Wounds and Injuries / physiopathology

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • CEACAM8 protein, human
  • Cell Adhesion Molecules
  • GPI-Linked Proteins
  • RNA
  • DNA