Clinical microfluidics for neutrophil genomics and proteomics.
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.
Kotz
Kenneth T
KT
Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA. kkotz@partners.org
Xiao
Wenzong
W
Miller-Graziano
Carol
C
Qian
Wei-Jun
WJ
Russom
Aman
A
Warner
Elizabeth A
EA
Moldawer
Lyle L
LL
De
Asit
A
Bankey
Paul E
PE
Petritis
Brianne O
BO
Camp
David G
DG
Rosenbach
Alan E
AE
Goverman
Jeremy
J
Fagan
Shawn P
SP
Brownstein
Bernard H
BH
Irimia
Daniel
D
Xu
Weihong
W
Wilhelmy
Julie
J
Mindrinos
Michael N
MN
Smith
Richard D
RD
Davis
Ronald W
RW
Tompkins
Ronald G
RG
Toner
Mehmet
M
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D019921:Biotinylation; D002056:Burns / Q000503:physiopathology*; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000276:immunology; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D058851:GPI-Linked Proteins; D023281:Genomics / Q000379:methods*; D006801:Humans; D044085:Microfluidics / Q000379:methods*; D009504:Neutrophils / Q000166:cytology / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D040901:Proteomics / Q000379:methods*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D014947:Wounds and Injuries / Q000503:physiopathology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; C504120:CEACAM8 protein, human; D015815:Cell Adhesion Molecules; D058851:GPI-Linked Proteins; D012313:RNA; D004247:DNA
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