In Vivo RNA Interference Screens Identify Regulators of Antiviral CD4+ and CD8+ T Cell Differentiation

被引:94
作者
Chen, Runqiang [1 ,2 ,3 ]
Belanger, Simon [2 ]
Frederick, Megan A. [3 ]
Li, Bin [2 ]
Johnston, Robert J. [2 ]
Xiao, Nengming [4 ]
Liu, Yun-Cai [4 ]
Sharma, Sonia [4 ]
Peters, Bjoern [2 ]
Rao, Anjana [1 ]
Crotty, Shane [2 ]
Pipkin, Matthew E. [3 ]
机构
[1] La Jolla Inst Allergy & Immunol, Div Signaling & Gene Express, La Jolla, CA 92037 USA
[2] La Jolla Inst Allergy & Immunol, Div Vaccine Discovery, La Jolla, CA 92037 USA
[3] Scripps Res Inst, Dept Canc Biol, Jupiter, FL 33458 USA
[4] La Jolla Inst Allergy & Immunol, Div Cell Biol, La Jolla, CA 92037 USA
关键词
FOLLICULAR HELPER; TRANSCRIPTION FACTORS; IMMUNE-RESPONSES; VIRAL-INFECTION; P-TEFB; MEMORY; EXPRESSION; ELONGATION; EFFECTOR; GENES;
D O I
10.1016/j.immuni.2014.08.002
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Classical genetic approaches to examine the requirements of genes for T cell differentiation during infection are time consuming. Here we developed a pooled approach to screen 30-100+ genes individually in separate antigen-specific T cells during infection using short hairpin RNAs in a microRNA context (shRNAmir). Independent screens using T cell receptor (TCR)-transgenic CD4+ and CD8+ T cells responding to lymphocytic choriomeningitis virus (LCMV) identifiedmultiple genes that regulated development of follicular helper (Tfh) and T helper 1 (Th1) cells, and short-lived effector and memory precursor cytotoxic T lymphocytes (CTLs). Both screens revealed roles for the positive transcription elongation factor (P-TEFb) component Cyclin T1 (Ccnt1). Inhibiting expression of Cyclin T1, or its catalytic partner Cdk9, impaired development of Th1 cells and protective short-lived effector CTL and enhanced Tfh cell and memory precursor CTL formation in vivo. This pooled shRNA screening approach should have utility in numerous immunological studies.
引用
收藏
页码:325 / 338
页数:14
相关论文
共 47 条
[1]   mTOR regulates memory CD8 T-cell differentiation [J].
Araki, Koichi ;
Turner, Alexandra P. ;
Shaffer, Virginia Oliva ;
Gangappa, Shivaprakash ;
Keller, Susanne A. ;
Bachmann, Martin F. ;
Larsen, Christian P. ;
Ahmed, Rafi .
NATURE, 2009, 460 (7251) :108-U124
[2]   Initial T cell receptor transgenic cell precursor frequency dictates critical aspects of the CD8+ T cell response to infection [J].
Badovinac, Vladimir P. ;
Haring, Jodie S. ;
Harty, John T. .
IMMUNITY, 2007, 26 (06) :827-841
[3]   Endogenous naive CD8+ T cell precursor frequency regulates primary and memory responses to infection [J].
Bar, Joshua J. ;
Khanna, Kamal M. ;
Lefrancois, Leo .
IMMUNITY, 2008, 28 (06) :859-869
[4]   RNAi screens in mice identify physiological regulators of oncogenic growth [J].
Beronja, Slobodan ;
Janki, Peter ;
Heller, Evan ;
Lien, Wen-Hui ;
Keyes, Brice E. ;
Oshimori, Naoki ;
Fuchs, Elaine .
NATURE, 2013, 501 (7466) :185-+
[5]   Transcriptional insights into the CD8+ T cell response to infection and memory T cell formation [J].
Best, J. Adam ;
Blair, David A. ;
Knell, Jamie ;
Yang, Edward ;
Mayya, Viveka ;
Doedens, Andrew ;
Dustin, Michael L. ;
Goldrath, Ananda W. .
NATURE IMMUNOLOGY, 2013, 14 (04) :404-412
[6]   The Cyclin K/Cdk12 complex maintains genomic stability via regulation of expression of DNA damage response genes [J].
Blazek, Dalibor ;
Kohoutek, Jiri ;
Bartholomeeusen, Koen ;
Johansen, Eric ;
Hulinkova, Petra ;
Luo, Zeping ;
Cimermancic, Peter ;
Ule, Jernej ;
Peterlin, B. Matija .
GENES & DEVELOPMENT, 2011, 25 (20) :2158-2172
[7]   Transcriptional regulator Id2 mediates CD8+ T cell immunity [J].
Cannarile, Michael A. ;
Lind, Nicholas A. ;
Rivera, Richard ;
Sheridan, Alison D. ;
Camfield, Kristin A. ;
Wu, Bei Bei ;
Cheung, Kitty P. ;
Ding, Zhaoqing ;
Goldrath, Ananda W. .
NATURE IMMUNOLOGY, 2006, 7 (12) :1317-1325
[8]   Asymmetric T lymphocyte division in the initiation of adaptive immune responses [J].
Chang, John T. ;
Palanivel, Vikram R. ;
Kinjyo, Ichiko ;
Schambach, Felix ;
Intlekofer, Andrew M. ;
Banerjee, Arnob ;
Longworth, Sarah A. ;
Vinup, Kristine E. ;
Mrass, Paul ;
Oliaro, Jane ;
Killeen, Nigel ;
Orange, Jordan S. ;
Russell, Sarah M. ;
Weninger, Wolfgang ;
Reiner, Steven L. .
SCIENCE, 2007, 315 (5819) :1687-1691
[9]   Bcl6 Expressing Follicular Helper CD4 T Cells Are Fate Committed Early and Have the Capacity To Form Memory [J].
Choi, Youn Soo ;
Yang, Jessica A. ;
Yusuf, Isharat ;
Johnston, Robert J. ;
Greenbaum, Jason ;
Peters, Bjoern ;
Crotty, Shane .
JOURNAL OF IMMUNOLOGY, 2013, 190 (08) :4014-4026
[10]   A Validated Regulatory Network for Th17 Cell Specification [J].
Ciofani, Maria ;
Madar, Aviv ;
Galan, Carolina ;
Sellars, MacLean ;
Mace, Kieran ;
Pauli, Florencia ;
Agarwal, Ashish ;
Huang, Wendy ;
Parkurst, Christopher N. ;
Muratet, Michael ;
Newberry, Kim M. ;
Meadows, Sarah ;
Greenfield, Alex ;
Yang, Yi ;
Jain, Preti ;
Kirigin, Francis K. ;
Birchmeier, Carmen ;
Wagner, Erwin F. ;
Murphy, Kenneth M. ;
Myers, Richard M. ;
Bonneau, Richard ;
Littman, Dan R. .
CELL, 2012, 151 (02) :289-303