title: Microfluidics for T- lymphocyte cell separation and inflammation monitoring in burn patients.
abstract: Severe burns result in T lymphocyte specific immunologic changes. In addition to decreased levels of circulating lymphocytes, changes in cytokine secretion and receptor expression also take place. Our finer understanding of the inflammatory response has led to the development of immune-targeted therapeutics, requiring specialized gene-expression monitoring. The emerging field of bio-micro-electromechanical systems can be used to isolate highly pure T lymphocytes in a clinically relevant and timely manner for downstream genomic analysis. Blood samples from healthy volunteers and burn-injured patients were introduced into microfluidic devices developed in our laboratory. Utilizing cell-affinity chromatography for positive selection of T lymphocytes, the devices served as a platform for RNA extraction and downstream cytokine analysis via quantitative real-time polymerase chain reaction (PCR). From a 0.5-mL whole blood sample, the microfluidic devices captured highly pure T lymphocytes from healthy volunteers and burn-injured patients. Cell capture was of sufficient quantity, and extracted RNA was of sufficient quality, for evaluating the gene expression of cytokines: interferon-gamma, interleukin-2, interleukin-4, and interleukin-10. Microfluidics is a useful tool in processing blood from burn-injured patients. Though in its very early stages of development, cell-specific information obtained by this platform/technology will likely be an important component of near-patient molecular diagnostics and personalized medicine.
authors.affiliation: Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Shriners Hospital for Children, Boston, Massachusetts, USA. alanrosenbach@hotmail.com
authors.forename: Alan E
authors.initials: AE
authors.lastname: Rosenbach
authors.forename: Piyush
authors.initials: P
authors.lastname: Koria
authors.forename: Jeremy
authors.initials: J
authors.lastname: Goverman
authors.forename: Kenneth T
authors.initials: KT
authors.lastname: Kotz
authors.forename: Amit
authors.initials: A
authors.lastname: Gupta
authors.forename: Ming
authors.initials: M
authors.lastname: Yu
authors.forename: Shawn P
authors.initials: SP
authors.lastname: Fagan
authors.forename: Daniel
authors.initials: D
authors.lastname: Irimia
authors.forename: Ronald G
authors.initials: RG
authors.lastname: Tompkins
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002056:Burns / Q000150:complications / Q000235:genetics* / Q000276:immunology*; D002469:Cell Separation; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007249:Inflammation / Q000150:complications / Q000235:genetics* / Q000276:immunology*; D008297:Male; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D008875:Middle Aged; D012313:RNA / Q000235:genetics / Q000592:standards; D013601:T-Lymphocytes / Q000166:cytology* / Q000378:metabolism; D018417:Th1 Cells / Q000378:metabolism; D018418:Th2 Cells / Q000378:metabolism; D055815:Young Adult
chemical_list: D016207:Cytokines; D012313:RNA