Main description:

This volume provides detailed descriptions on cutting-edge experimental methods to assess the multiple and complex interactions between T. cruzi and its host. Chapters detail a series of methods and protocols, ranging from rather classical biochemical/genetic approaches to modern, powerful and high-throughput '-omics' technologies, to tackle this issue, as well as novel techniques to improve treatment and clinical evaluation of Chagasic patients. Genome-wide mining strategies aimed at identifying potential parasite antigens and vaccine candidates and an overview on the challenges and milestones encountered during T. cruzi genome assembly are also included. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, T. cruzi Infection: Methods and Protocols aims to provide students, researchers, and clinicians who are interested in the study of Chagas disease an indispensable source of information.

Contents:

Preface...Table of Contents...Contributing Authors...
1. Trypanosoma cruzi Genome Assemblies: Challenges and Milestones of Assembling a Highly Repetitive and Complex GenomeJoao Luis Reis-Cunha and Daniella C. Bartholomeu
2. Mining Trypanosoma cruzi Genome Sequences for Antigen Discovery and Vaccine DevelopmentChristian Teh-Poot and Eric Dumonteil
3. Transcriptome Studies in Trypanosoma cruzi using RNAseqGonzalo Greif, Luisa Berna, Florencia Diaz-Viraque, and Carlos Robello
4. An Improved Approach to Trypanosoma cruzi Molecular Genotyping by Next-Generation Sequencing of the Mini-Exon GeneAlicia Majeau, Claudia Herrera, and Eric Dumonteil
5. Genome Editing by CRISPR/Cas9 in Trypanosoma cruziNoelia Lander, Miguel A. Chiurillo, and Roberto Docampo
6. Sample Preparation of Trypanosoma cruzi Surface, Extracellular, and Nuclear Subproteomes Carlos Andre Ornelas Ricart, Rayner Myr Lauterjung Queiroz, Agenor de Castro Moreira dos Santos Junior, and Sebastien Charneau
7. Isolation and Characterization of Extracellular Vesicles Derived from Trypanosoma cruziIzadora Volpato Rossi, Bruno Gavinho, and Marcel Ivan Ramirez
8. Production of Recombinant Trypanosoma cruzi Antigens in Leishmania tarentolae Maria Jose Ferrer, Diana Patricia Wehrendt, Mariana Bonilla, Marcelo Alberto Comini, Maria Teresa Tellez-Inon, and Mariana Potenza 9. Homologous Expression of Glycosylphosphatidyl Inositol-Anchored Glycoproteins in Trypanosoma cruzi Virginia Balouz, Andrea C. Mesias, Camila Centeno Camean, Ivana Ducrey, Maite Mabel Lobo, Ignacio M. Durante, Gaspar E. Canepa, Carlos A. Buscaglia, and Maria de los Milagros Camara
10. Metabolic Labeling of Surface Neo-sialylglyconjugates Catalyzed by Trypanosoma cruzi trans-SialidaseGiannina Carlevaro, Andres B. Lantos, Gaspar E. Canepa, Maria de los Milagros Camara, Martin Somoza, Carlos A. Buscaglia, Oscar Campetella, and Juan Mucci
11. Exploiting Genetically Modified Dual-Reporter Strains to Monitor Experimental Trypanosoma cruzi Infections and Host: Parasite Interactions Martin C. Taylor, Amanda F. Francisco, Shiromani Jayawardhana, Gurdip Singh Mann, Alexander Ward, Francisco Olmo, Michael D. Lewis, and John M. Kelly
12. Transmigration of Trypanosoma cruzi trypomastigotes through 3D Spheroids Mimicking Host TissuesMatias Exequiel Rodriguez, Mariana Rizzi, Lucas Caeiro, Yamil Masip, Daniel O. Sanchez, and Valeria Tekiel
13. In Situ Detection of Dormant Trypanosoma cruzi Amastigotes using Bioluminescent-Fluorescent ReportersFernando Sanchez-Valdez and Angel Padilla
14. Multi-Parametric Evaluation of Trypanosoma cruzi Infection Outcome in Animal ModelsJulien Santi-Rocca, Nuria Girones, and Manuel Fresno
15. MicroRNA Transcriptome Profiling in Heart of Trypanosoma cruzi-Infected MiceLudmila Rodrigues Pinto Ferreira
16. Genotyping of Trypanosoma cruzi from Clinical Samples by Multilocus Conventional PCR Otacilio C. Moreira and Juan Carlos Ramirez
17. Assessing Parasite Load in Chagas Disease Patients by Quantitative Multiplex Real-Time PCRJuan Carlos Ramirez and Otacilio da Cruz Moreira
18. Diagnostic Applicability of Neutralizing Antibodies to Trypanosoma cruzi trans-sialidase Leguizamon Maria Susana
19. Analytics of Functional Autoantibodies in Patients with Chagas' DiseaseGerd Wallukat, Katrin Wenzel, and Ingolf Schimke
20. Apolipoprotein-A1 and Fibronectin Fragments as Markers of Cure for the Chagas DiseaseElizabeth Ruiz-Lancheros, Makan Golizeh, and Momar Ndao
21. Host Derived Molecules as Novel Chagas Disease Biomarkers: Hypercoagulability Markers in PlasmaJulio Alonso-Padilla, Dolors Tassies, Nuria Cortes-Serra, Joaquim Gascon, Joan-Carles Reverter, and Maria-Jesus Pinazo
22. Purification of GlycosylphosphatidyLinositol-Anchored Mucins from Trypanosoma cruzi Trypomastigotes and Synthesis of -Gal-Containing Neoglycoproteins: Application as Biomarkers for Reliable Diagnosis and Early Assessment of Chemotherapeutic Outcomes of Chagas DiseaseUriel Ortega-Rodriguez, Susana Portillo, Roger A. Ashmus, Jerry A. Duran, Nathaniel S. Schocker, Eva Iniguez , Alba L. Montoya, Brenda G. Zepeda, Janet J. Olivas, Nasim H. Karimi, Julio Alonso-Padilla, Luis Izquierdo, Maria-Jesus Pinazo, Belkisyole Alarcon de Noya , Oscar Noya, Rosa A. Maldonado, Faustino Torrico, Joaquim Gascon , Katja Michael, and Igor C. Almeida
23. Cytometric Bead Array (CBA) for Measurement Cytokines Levels in Chagas Disease PatientsNayara I. Medeiros and Juliana A. S. Gomes
24. Generation of T. cruzi-specific Primary CD4+ T Cell Lines from Peripheral Blood Mononuclear Cells Isolated from Chagas Disease Patients Gonzalo R. Acevedo, Paula B. Alcaraz, Clemencia Pinilla, and Karina A. Gomez
25. Trypanosma cruzi Specific T-Cell Responses to Monitor Treatment Efficacy in Chronic Chagas DiseaseMaria Cecilia Albareda, Maria Ailen Natale, Gonzalo Leandro Cesar, Melisa Daiana Castro Eiro, Maria Gabriela Alvarez, and Susana Adriana Laucella
26. CD8+ T Cell Response to Trypanosoma cruzi Antigens during Chronic Chagas DiseasePaola Lasso, Jose Mateus, John Mario Gonzalez, Adriana Cuellar, and Concepcion Puerta
27. Fluorescence Proteomic Technology to Analyze Peripheral Blood Mononuclear Cells in Chronic Chagas Disease John E. Wiktorowicz, M. Paola Zago, and Nisha J. Garg
28. Isolation and Phenotypic Characterization of Inflammatory Cells from Clinical Samples: Purification of Macrophages from Trypanosoma cruzi-Infected HeartsNatalia Eberhardt, Liliana Maria Sanmarco, and Maria Pilar Aoki