Cryptococcus neoformans, 1st ed. 2024 Methods and Protocols Methods in Molecular Biology Series, Vol. 2775

This volume explores the latest developments in the study of Cryptococcus neoformans and its pathology, along with discussion on newly used therapeutics. The chapters in this book cover a wide range of protocols commonly used in Cryptococcus research such as animal models, genetics techniques, virulence factor phenotyping, and microscopy. 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.

Cutting-edge and thorough, Cryptococcus neoformans: Methods and Protocols is a valuable resource that will help researchers further enhance their understanding of this pathogenic yeast, and will aid in new discoveries and therapeutics.

Preface…
Table of Contents…
Contributing Authors…

Part I Virulence Models

1.     A Galleria Model of Cryptococcus neoformans Infection
Helene C. Eisenman

2.     The Rabbit Model of Cryptococcal Meningitis
Charles Giamberardino and John Perfect

3.     Models for Inducing Experimental Cryptococcosis in Mice
Natalia Castro-Lopez and Floyd L. Wormley Jr.

4.     In Vivo Modeling of Cryptococcus neoformans Infection and Collection of Murine Samples
Ben Muselius and Jennifer Geddes-McAlister

Part II Transformation

5.     Biolistic Transformation of Cryptococcus neoformans
Dena L. Toffaletti, Jennifer L. Tenor, and John R. Perfect

6.     Genetic Transformation of Cryptococcus Species with Agrobacterium Transfer DNA
Nicholas F. Chong, Alexander Idnurm, and Bridgit C. Nugent

7.     Gene Silencing via RNA Interference in Cryptococcus
Indrani Bose

Part III ‘Omics

8.     Cryptococcus neoformans Transcriptome Analysis to Identify Differentially Expressed Genes Using the STAR Pipeline in CyVerse Discovery Environment
Rebecca L. Seipelt-Thiemann

9.     Proteomic Profiling of Samples Derived from a Murine Model Following Cryptococcus neoformans Infection
Ben Muselius, Antoine Bodein, Florence Roux-Dalvai, Arnaud Droit, and Jennifer Geddes-McAlister

Part IV Microscopy

10.  Electron Microscopy of Cryptococcus neoformans: Processing Challenges to Avoid Artifacts Glauber R. de S. Araújo, Bruno Pontes, and Susana Frases

Part V Interactions with Macrophages

11.  Assessing Phagocytosis of Cryptococcus neoformans Cells in Human Monocytes or the J774 Murine Macrophage Cell Line
Emmanuel Lafont, Aude Sturny-Leclère, Carolina Coelho, Fanny Lanternier, and Alexandre Alanio

12.  Interaction Between Macrophages and Cryptococcus neoformans: Distinguishing Phagocytosed Versus External Fungi
Amanda Pereira Rocha, Thaís Silva Alves, Adrielle Veloso Caixeta, Patrícia Albuquerque, and André Moraes Nicola

13.  Detection and Quantification of Cryptococcus Uptake by Phagocytic Cells Using Imaging Flow Cytometry
Althea Campuzano, Chiung-Yu Hung, and Floyd L. Wormley Jr.

14.  Two Methods of Measuring Cryptococcus neoformans Fungal Burden in Macrophages
Emily Smith-Peavler, Linda M. Sircy, David E. Nelson, and Erin E. McClelland

Part VI Virulence Factors

15.  Quantification of C. neoformans Capsule Diameter
Tiffany E. Guess, Hoyin Lai, David E. Nelson, and Erin E. McClelland

16.  Methods of Cryptococcal Polysaccharide Analysis Using ELISA
Maggie P. Wear, Scott A. McConnell, Seth D. Greengo, Livia Liporagi Lopes, and Arturo Casadevall

17.  Measuring Laccase Activity and Melanin Production in Cryptococcus neoformans
Patrícia Albuquerque, Herdson Renney de Sousa, Stefânia de Oliveira Frazão, Larissa Verginia do Nascimento Miranda, Hugo Costa Paes, Ildinete Silva Pereira, and André Moraes Nicola

18.  Measuring Urease and Phospholipase Secretion in Cryptococcus neoformans
Hunter J. Hoffman and Erin E. McClelland

19.  Measuring Stress Phenotypes in Cryptococcus neoformans
Rajendra Upadhya, Corinna Probst, J. Andrew Alspaugh, and Jennifer K. Lodge

Part VII Measuring Specific Phenotypes

20.  Antibody Isolation in C. neoformans
Susana Ruiz-Mendoza, Marina da Silva Ferreira, Michele Ramos Valente, and Allan J. Guimarães

21.  Fluorescence and Biochemical Assessment of the Chitin and Chitosan Content of Cryptococcus Brian T. Maybruck, Rajendra Upadhya, Woei C. Lam, Charles A. Specht, andJennifer K. Lodge

22.  Induction of Dormancy in Cryptococcus neoformans In Vitro: The HypNOS Protocol
Ruchi Agrawal, Aude Sturny-Leclère, Raffael J. A. de Castro, and Alexandre Alanio

23.  Analysis of Cryptococcus Extracellular Vesicles
Flavia C. G. Reis, Rafael F. Castelli, Diogo Kuczera, Haroldo C. Oliveira, and Marcio L. Rodrigues

24.  Preparation of Biologically Active Fractions Enriched with Glucuronoxylomannan, the Main Antigen of the Cryptococcal Capsule
Luisa J. Jozefowicz and Marcio L. Rodrigues

25.  Measuring Replicative Lifespan in Cryptococcus neoformans
Vanessa K.A. Silva, Natalia K. Oliveira, and Bettina C. Fries

26.  In Vitro Titan Cell Generation in Cryptococcus neoformans and Automated Cell Size Measurements
Benjamin Hommel, Aude Sturny-Leclère, and Alexandre Alanio

27.  Chitosan-Deficient Cryptococcus as Whole-Cell Vaccines
Charles A. Specht, Woei C. Lam, Maureen M. Hester, Diana Lourenco, Stuart M. Levitz, Jennifer K. Lodge, and Rajendra Upadhya

28.  Vaccine Strategies for Cryptococcus neoformans
Veronica S. Brauer, Samyr Machado Querobino, Gabriel S. Matos, Deveney Dasilva, and Maurizio Del Poeta

Subject Index List…

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts