UC San Diego UC San Diego Electronic Theses and Dissertations Title The role of Erk MAPK pathway in CD4 T cell proliferation and differentiation Permalink https://escholarship.org/uc/item/6tb5b89q Author Chang, Chiung-Fang Publication Date 2011 Peer reviewed|Thesis/dissertation eScholarship.org Powered by the California Digital Library University of California UNIVERSITY OF CALIFORNIA, SAN DIEGO The role of Erk MAPK pathway in CD4 T cell proliferation and differentiation A dissertation submitted in partial satisfaction of the requirements for the degree Doctor of Philosophy in Biology by Chiung-Fang Chang Committee in charge: Professor Stephen M. Hedrick, Chair Professor Ananda W. Goldrath Professor Cornelis Murre Professor David Traver Professor Carl F. Ware Professor Maurizio Zanetti 2011 Copyright Chiung-Fang Chang, 2011 All rights reserved The Dissertation of Chiung-Fang Chang is approved, and it is acceptable in quality and form for publication on microfilm and electronically: Chair University of California, San Diego 2011 iii DEDICATION To my parents, for their encouragement and support iv TABLE OF CONTENTS SIGNATURE PAGE ............................................................................................ iii DEDICATION ...................................................................................................... iv TABLE OF CONTENTS ....................................................................................... v LIST OF FIGURES ............................................................................................ vii LIST OF TABLES ............................................................................................... ix ACKNOWLEDGEMENTS .................................................................................... x VITA ................................................................................................................... xii ABSTRACT OF THE DISSERTATION ............................................................. xiv Chapter. 1 General introduction ........................................................................... 1 MAPK (mitogen activated protein kinase) pathway ......................................... 2 ERK1/2 and their substrates ............................................................................ 3 T cell activation and differentiation ................................................................... 6 Immune responses .......................................................................................... 8 CD4 T cell differentiation .................................................................................. 9 Autoimmune diseases and CD4 T cells ......................................................... 14 Summary ........................................................................................................ 15 Chapter. 2 Phenotype of Erk2f/f CreERT2 mice and CD4 T cell activation ......... 20 Abstract .......................................................................................................... 21 Introduction .................................................................................................... 21 Results ........................................................................................................... 25 Discussion ...................................................................................................... 29 Experimental procedures ............................................................................... 30 Chapter. 3 The role of Erk in CD4 T helper proliferation and differentiation ...... 40 v Abstract .......................................................................................................... 41 Introduction .................................................................................................... 41 Results ........................................................................................................... 42 Discussion ...................................................................................................... 45 Experimental procedures ............................................................................... 46 Chapter. 4 The role of Erk in induced regulatory T proliferation and differentiation ..................................................................................................... 62 Abstract .......................................................................................................... 59 Introduction .................................................................................................... 59 Results ........................................................................................................... 61 Discussion ...................................................................................................... 64 Experimental procedures ............................................................................... 65 Chapter. 5 Microarray analysis for the role of Erk2 in Th1 and iTreg ................ 76 Abstract .......................................................................................................... 77 Introduction .................................................................................................... 77 Results ........................................................................................................... 78 Discussion ...................................................................................................... 94 Experimental procedures ............................................................................... 95 Chapter. 6 General discussion ........................................................................ 115 References ...................................................................................................... 126 vi LIST OF FIGURES Figure 1-1. T cells activation and proliferation ................................................... 17 Figure 1-2. T cell receptor signaling and Erk target genes. ............................... 18 Figure 1-3. Differentiation of T helper cells and regulatory T cells. ................... 19 Figure 2-1. Erk2f/fCreERT2 mice. ........................................................................ 33 Figure 2-2. Deletion of Erk2 in Erk2f/f Cre-ERT2 mice following tamoxifen treatment ......................................................................................... 34 Figure 2-3. Secondary lymphoid tissues of Erk2f/f Cre-ERT2 mice appear normal but there is a defect in Erk2f/f Cre-ERT2 thymus. ............................. 35 Figure 2-4. Erk1 deficient CD4 T cells exhibit normal proliferation under stimulation ....................................................................................... 36 Figure 2-5. Erk2 deficient CD4 T cells have impaired proliferation with TCR signaling alone but exhibit normal proliferation under optimal stimulation in vitro ........................................................................... 37 Figure 2-6. Erk2-deficient Smarta CD4 T cells have impaired proliferation with GP peptide alone but exhibit normal proliferation under optimal stimulation in vitro ........................................................................... 38 Figure 2-7. Erk deficient CD4 T cells have impaired survival and proliferation under optimal stimulation in vitro .................................................... 39 Figure 3-1. Normal proliferation of Erk1 deficient CD4 T cells ........................... 48 Figure 3-2. Erk1 deficient CD4 T cells exhibit normal CD4 differentiations. ...... 49 Figure 3-3. Erk2-/- CD4 T cells display impaired survival to the Th1 subset ...... 50 Figure 3-5. Erk2-/- CD4 T cells exhibit impaired polarization to the Th1 subset . 52 Figure 3-6. Erk2-deficient CD4 T cells express normal T-bet and phos-stat-1, but more Foxp3, Gata3 under Th1 skewing condition. condition .... 53 Figure 3-7. Erk1-/-2-/- CD4 T cells display impaired survival to the Th1 subset .. 54 Figure 3-8. Erk1-/-2-/- CD4 T cells impaired polarization to the Th1 subset. ....... 55 Figure 3-9. Erk2 deficient mice have the impaired anti-viral Th1 responses. .... 56 Figure 3-10. Impaired anti-viral Th1 responses in absence of Erk2 .................. 57 Figure 4-1. Slightly increased percentages and cell number in Erk2f/fCD4Cre lymph nodes and spleens ............................................................... 69 Figure 4-2. Slightly increased nTreg percentages and normal nTreg cell number in Erk2f/fERCre mice ....................................................................... 70 Figure 4-3. Enhanced differentiation of induced Treg in absence of Erk2. ........ 71 Figure 4-4. Enhanced differentiation of induced Treg in absence of total Erk (Erk1 and Erk2) and Erk2, but not Erk1 alone. ............................... 72 Figure 4-5. In vivo enhanced differentiation of induced Treg without Erk2 ........ 73 Figure 4-6. Normal function of Treg in absence of Erk2 .................................... 74 Figure 4-7. Erk2 regulate DNMT and phos-Smad under iTreg condition. .......... 75 Figure 5-1. Experiment Design for microarray ................................................... 97 vii Figure 5-2. Day 0 scatter plot for the expression of WT and Erk2-/- CD4 T cells98 Figure 5-3. Scatters plots show that the mean expression of wild type (WT) CD4 T cells and Erk2-/- CD4 T cells under different conditions ............... 99 Figure 5-4. Scatters plots show that the mean expression wild type (WT) CD4 T cells and Erk2-/- CD4 T cells under different