Tumors and Ly6Chigh Monocytes Fatemeh Zare

Tumors and Ly6Chigh Monocytes
Fatemeh Zare
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PhD Programme in Biomedicine
Programa de Doctorado en Biomedicina
Tumors and Ly6Chigh Monocytes
Tumores y Monocitos Ly6Chigh
Thesis submitted by - Memoria presentada por
Fatemeh Zare
To qualify for the Doctorate degree by - Para optar al grado de Doctor por la
University of Barcelona - Universidad de Barcelona

Programa de Doctorado de Biomedicina
Bienio 2014-2015
Memoria presentada por
Fatemeh Zare
Para optar al Grado de Doctora por la
Universidad de Barcelona
Director de Tesis
Dr. Antonio Celada Cotarelo
Catedratico de Inmunologia
Barcelona, Octubre de 2014
Departamento de Fisiologia y Inmunologia
Facultat de Biologia
Universidad de Barcelona

ACKNOWLEDGMENTS
My PhD has been a lengthy journey and many people have helped
me along the way. Firstly I would like to acknowledge my supervisor for all
his help and support throughout this time. Prof. Antonio Celada, thank you
for encouraging me to work independently as a scientist and for providing
me the opportunity to be a part of your group. Prof Roger Gomis, given that
for you I was an external student, I wish to especially thank you for your
constant support. Dr. Jordi Lloberas, thank you for your wonderful problem
solving skills, especially under pressure, during those times when I showed
up at your office door with a million questions. My gratitude also goes to Dr.
Jaume Comas for teaching me flow cytometry, and developing the cell
sorting protocol. I would like to extend my appreciation to all the members of
the Macrophage Biology, both past and present, for their technical support
and encouragement. Thank you to Juan A. calatayud for our rewarding
collaboration and for the many discussions about our work. A million thanks
to Dr. Gloria Sans, Dr. Selma Kuznetsova, Lorena V Strella, Juan Torres and
Ester Ruiz that I could count on your help always. Many thanks to our lab
technician Gemma López for her unending patience and help around the
laboratory. I would like to thank my family and friends for their love and
support during my studies. I am eternally grateful to my wonderful parents
and sister who have always encouraged me in everything and who have
been there not only for all the highs , but importantly for all the lows too.
Without you I would never have gotten this far. This is for you.

TABLE OF CONTENTS
1 INTRODUCTION ...................................................................................... 14
1.1 Cancer .................................................................................................. 15
1.1.1 Cancer and the Immune System .................................................. 16
1.1.2 Tumor Immunology ....................................................................... 16
1.1.3 Cancer Related Inflammation (CRI) ............................................. 17
1.2 Innate and adaptive immune response to tumors ................................ 18
1.3 Immune cells and anti-cancer therapies ............................................... 21
1.4 Cells in the tumor microenvironment .................................................... 23
1.4.1 Inflammatory cell component of tumors ....................................... 24
1.4.2 Tumor-associated leukocytes ....................................................... 25
1.4.3 Macrophages ................................................................................ 26
1.4.3.1 Tumor associated macrophages .............................................. 26
1.4.3.1.1 TAMs phenotype ................................................................ 27
1.4.4 Monocytes .................................................................................... 32
1.4.4.1 Monocyte heterogeneity ........................................................... 33
1.4.4.2 Monocytes sub populations ...................................................... 35
1.4.4.3 Recruitment of monocytes to sites of inflammation ................. 38
1.4.4.4 Ly6Chigh and Ly6Clow monocytes .............................................. 39
1.4.4.5 Monocytes differentiation to macrophages .............................. 42
1.4.4.6 Monocytes recruitment to tumors ............................................. 43
1.5 Monocytes generation .......................................................................... 46
1.6 Animal models ...................................................................................... 48
1.7 Objectives ............................................................................................. 48
2 MATERIALS AND METHODS.................................................................. 49
2.1 Mouse strains ....................................................................................... 50
2.2 Cell lines and culture ............................................................................ 50
2.2.1 MDA-MB-231 cell line ................................................................... 50
2.2.2 C26 cell line .................................................................................. 51