The study of host-pathogen interaction is pivotal to understand the pathogenesis of
leptospirosis. Motility and chemotaxis of leptospires are essential for rapid dissemination in
the host. An in vitro assay for the identification of chemoattractants is included in this
volume (Chapter 11). Protocols for in situ structural analysis of periplasmic flagella in
Leptospira spp. by electron cryotomography (Chapter 12) and measurement of cell-body
rotation (Chapter 13) are provided. Bioluminescence imaging can be used to follow bacterial dissemination in a mouse in real time and in a noninvasive way (Chapter 14), and
translocation of bacteria across cell monolayers can be determined (Chapter 15). After
entering bloodstream, leptospires can resist host complement system, bind to host cells
and the extracellular matrix, and evade host defense mechanism in macrophages. In this
book, in vitro/ex vivo assays for the identification of leptospiral adhesion proteins
(Chapter 16), leptospiral complement resistance (Chapter 17), and intracellular trafficking
of leptospires in macrophages (Chapter 18) are included. Biofilm formation probably
enhances the survival of Leptospira in the environment, and a protocol for quantification
of biofilms is also included (Chapter 19). Assays for the measurement of viability of
leptospires are also described (Chapter 20).
This book also offers protocols for in vivo analysis of transcriptional/proteomic changes
of L. interrogans using dialysis membrane chamber implanted in the peritoneal cavity of rats
(Chapter 21). Quantification of bacterial burden and host cytokine gene expression in the
hamster model of acute leptospirosis is also described in Chapter 22. Hamsters are also used
to evaluate vaccine candidates for leptospirosis (Chapter 23).
In addition to experimental protocols, this book includes review articles providing an
overview of Leptospira and leptospirosis (Chapter 24) and laboratory diagnosis of leptospirosis (Chapter 25).
This book thus summarizes all aspects of manipulation of Leptospira spp., from strain
isolation to cutting-edge approaches for studying the pathogenesis of leptospirosis. This
book reflects the progress that has been made in the field in the last 10 years. All the
contributors are leading researchers in the Leptospira and leptospirosis field, and we would
like to thank them for providing their comprehensive protocols and techniques for this
book. We also would like to thank Dr. John Walker, the Series Editor of Methods in
Molecular Biology, for giving us the opportunity to edit this book and his continuous
support.
We hope that this book will help new researchers join the field and study these fastidious
but fascinating organisms.
Tokyo, Japan
Nobuo Koizumi
Paris, France
Mathieu Picardeau
vi
Preface
leptospirosis. Motility and chemotaxis of leptospires are essential for rapid dissemination in
the host. An in vitro assay for the identification of chemoattractants is included in this
volume (Chapter 11). Protocols for in situ structural analysis of periplasmic flagella in
Leptospira spp. by electron cryotomography (Chapter 12) and measurement of cell-body
rotation (Chapter 13) are provided. Bioluminescence imaging can be used to follow bacterial dissemination in a mouse in real time and in a noninvasive way (Chapter 14), and
translocation of bacteria across cell monolayers can be determined (Chapter 15). After
entering bloodstream, leptospires can resist host complement system, bind to host cells
and the extracellular matrix, and evade host defense mechanism in macrophages. In this
book, in vitro/ex vivo assays for the identification of leptospiral adhesion proteins
(Chapter 16), leptospiral complement resistance (Chapter 17), and intracellular trafficking
of leptospires in macrophages (Chapter 18) are included. Biofilm formation probably
enhances the survival of Leptospira in the environment, and a protocol for quantification
of biofilms is also included (Chapter 19). Assays for the measurement of viability of
leptospires are also described (Chapter 20).
This book also offers protocols for in vivo analysis of transcriptional/proteomic changes
of L. interrogans using dialysis membrane chamber implanted in the peritoneal cavity of rats
(Chapter 21). Quantification of bacterial burden and host cytokine gene expression in the
hamster model of acute leptospirosis is also described in Chapter 22. Hamsters are also used
to evaluate vaccine candidates for leptospirosis (Chapter 23).
In addition to experimental protocols, this book includes review articles providing an
overview of Leptospira and leptospirosis (Chapter 24) and laboratory diagnosis of leptospirosis (Chapter 25).
This book thus summarizes all aspects of manipulation of Leptospira spp., from strain
isolation to cutting-edge approaches for studying the pathogenesis of leptospirosis. This
book reflects the progress that has been made in the field in the last 10 years. All the
contributors are leading researchers in the Leptospira and leptospirosis field, and we would
like to thank them for providing their comprehensive protocols and techniques for this
book. We also would like to thank Dr. John Walker, the Series Editor of Methods in
Molecular Biology, for giving us the opportunity to edit this book and his continuous
support.
We hope that this book will help new researchers join the field and study these fastidious
but fascinating organisms.
Tokyo, Japan
Nobuo Koizumi
Paris, France
Mathieu Picardeau
vi
Preface