title: A clinical microchip for evaluation of single immune cells reveals high functional heterogeneity in phenotypically similar T cells.
abstract: Cellular immunity has an inherent high level of functional heterogeneity. Capturing the full spectrum of these functions requires analysis of large numbers of effector molecules from single cells. We report a microfluidic platform designed for highly multiplexed (more than ten proteins), reliable, sample-efficient (∼1 × 10(4) cells) and quantitative measurements of secreted proteins from single cells. We validated the platform by assessment of multiple inflammatory cytokines from lipopolysaccharide (LPS)-stimulated human macrophages and comparison to standard immunotechnologies. We applied the platform toward the ex vivo quantification of T cell polyfunctional diversity via the simultaneous measurement of a dozen effector molecules secreted from tumor antigen-specific cytotoxic T lymphocytes (CTLs) that were actively responding to tumor and compared against a cohort of healthy donor controls. We observed profound, yet focused, functional heterogeneity in active tumor antigen-specific CTLs, with the major functional phenotypes quantitatively identified. The platform represents a new and informative tool for immune monitoring and clinical assessment.
authors: NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, California, USA.
authors: Chao
authors: C
authors: Ma
authors: Rong
authors: R
authors: Fan
authors: Habib
authors: H
authors: Ahmad
authors: Qihui
authors: Q
authors: Shi
authors: Begonya
authors: B
authors: Comin-Anduix
authors: Thinle
authors: T
authors: Chodon
authors: Richard C
authors: RC
authors: Koya
authors: Chao-Chao
authors: CC
authors: Liu
authors: Gabriel A
authors: GA
authors: Kwong
authors: Caius G
authors: CG
authors: Radu
authors: Antoni
authors: A
authors: Ribas
authors: James R
authors: JR
authors: Heath
references: Clin Cancer Res. 2009 Oct 1;15(19):6267-76
references: 19789309
references: Nat Rev Immunol. 2003 Dec;3(12):931-9
references: 14647475
references: J Am Chem Soc. 2009 Jul 22;131(28):9695-703
references: 19552409
references: Immunity. 2007 Sep;27(3):393-405
references: 17892848
references: Nat Med. 2004 Aug;10(8):806-10
references: 15286782
references: Immunobiology. 2004;209(1-2):191-8
references: 15481153
references: J Am Chem Soc. 2007 Feb 21;129(7):1959-67
references: 17260987
references: Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14863-8
references: 9843981
references: Nature. 2002 Dec 19-26;420(6917):860-7
references: 12490959
references: Cancer Cell. 2007 Oct;12(4):300-1
references: 17936555
references: Nat Immunol. 2008 May;9(5):450-3
references: 18425095
references: Chemphyschem. 2010 Oct 4;11(14):3063-9
references: 20715281
references: Nat Med. 2003 Mar;9(3):315-21
references: 12592402
references: Nat Biotechnol. 2008 Dec;26(12):1373-8
references: 19029914
references: Cancer Res. 2009 Nov 1;69(21):8412-9
references: 19843860
references: Nat Rev Immunol. 2008 Apr;8(4):247-58
references: 18323851
references: Nature. 2008 Jul 24;454(7203):428-35
references: 18650913
references: Cytometry A. 2008 Nov;73(11):975-83
references: 18785267
references: Clin Cancer Res. 2006 Jan 1;12(1):107-16
references: 16397031
references: Nature. 2000 Aug 17;406(6797):782-7
references: 10963608
references: Biophys J. 2011 May 18;100(10):2378-86
references: 21575571
references: Nat Med. 1999 Jun;5(6):677-85
references: 10371507
references: Nature. 2008 Jul 24;454(7203):436-44
references: 18650914
references: Nat Med. 2005 Oct;11(10):1113-7
references: 16186817
references: Springer Semin Immunopathol. 2006 Nov;28(3):209-19
references: 16932955
references: Cancer Cell. 2008 Oct 7;14(4):335-43
references: 18835035
references: Nat Med. 2007 Jul;13(7):843-50
references: 17558415
references: Immunol Rev. 2009 Jan;227(1):264-82
references: 19120490
references: J Exp Med. 2007 Jun 11;204(6):1405-16
references: 17535971
references: Proc Natl Acad Sci U S A. 2005 May 31;102(22):7916-21
references: 15905333
references: Science. 2006 Oct 6;314(5796):126-9
references: 16946036
references: J Transl Med. 2008 May 01;6:22
references: 18452610
references: Nat Rev Immunol. 2005 Dec;5(12):953-64
references: 16322748
references: Lab Chip. 2010 Nov 21;10(22):3157-62
references: 20924527
references: Science. 2002 Oct 18;298(5593):580-4
references: 12351675
references: J Immunol. 2005 Oct 1;175(7):4686-96
references: 16177116
references: J Immunol. 2004 Nov 1;173(9):5372-80
references: 15494483