Main description:

This second edition provides new and updated chapters useful for the study of Regulatory B cells. Organized in four sections, chapters detail basic methods for the isolation and immunophenotypical analysis of these cells, experimental approaches for the ex vivo generation/expansion of IL-10 producing B cells, molecular biology techniques for the analysis of IL-10 expression and production, and animal models mimicking pathologic settings. Written in the highly successful Methods in Molecular Biology series format, chapters include an introduction to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Regulatory B Cells: Methods and Protocols, Second Edition aims to be useful to the scientific community and serve to clarify some unsolved aspects of Regulatory B Cells research.

Contents:

Part I: Purification and Phenotypic Characterization of B Cell Subsets with Breg Traits

1. Purification and characterization of murine MZ and T2-MZP cells M. Manuela Rosado, Alaitz Aranburu, Marco Scarsella, Simona Cascioli, Ezio Giorda, and Rita Carsetti

2. Purification and immune-phenotyping of B-1 cells from body cavities of mice Vanessa Yenson and Nicole Baumgarth

3. Purification and immunophenotypic characterization of murine plasma cells Van Duc Dang, Simon Fillatreau and Andreia C. Lino

4. Purification of murine and human IL-10 producing B cells from different anatomical compartments Francesca Mion, Eleonora Martinis, Carlo E.M. Pucillo and Silvia Tonon

5. Purification and immunophenotypic characterization of human CD19+CD24hiCD38hi and CD19+CD24hiCD27+ B cells Hannah F. Bradford and Claudia Mauri

Part II: Mechanisms of Immune Suppression by B Cells

6. Detection of IL-10 in murine B cells: in vitro and in vivo techniques Xiang Lin, Xiaohui Wang and Liwei Lu

7. Detection and quantification of transforming growth factor- 1 produced by murine B cells: pros and cons of different techniques Yoshiyuki Mishima, Akihiko Oka and Shunji Ishihara

8. IL-35 detection in B cells at the mRNA and protein level Bhalchandra Mirlekar, Daniel Michaud and Yuliya Pylayeva-Gupta

9. Characterization and activation of Fas Ligand producing mouse B cells and their killer exosomes Steven K. Lundy, Sophina H. Taitano, and Lucien E.P.M. van der Vlugt

10. Characterization and activity of TIM-1 and IL-10-reporter expressing regulatory B cells Kanishka Mohib, David M. Rothstein and Qing Ding

11. New method for the expansion of highly purified human regulatory granzyme B-expressing B cells Melanie Chesneau, Hoa L. Mai and Sophie Brouard

12. Characterization of the cell surface phenotype and regulatory activity of B cell IgD low (BDL) regulatory B cells Mohamed Khalil, Avijit Ray and Bonnie N. Dittel

Part III: Methods for the ex vivo expansion and molecular characterization of IL-10 producing B cells

13. Use of Toll-like receptor (TLR) ligation to characterize human regulatory B cells subsets Mathilde A.M. Chaye, Chiara Tontini, Arifa Ozir-Fazalalikhan, Astrid L. Voskamp and Hermelijn H. Smits

14. Use of co-cultures for the study of cellular interactions influencing B cell regulatory functions Giada Pace and Viviana Valeri

15. Use of inhibitory compounds to dissect the molecular pathways involved in regulatory B cell differentiation Maik Luu and Alexander Visekruna

16. Methods to study the transcriptome of regulatory B cells Maud Maho-Vaillant and Sebastien Calbo

17. Purification of murine IL-10+ B cells for analyses of biological functions and transcriptomics Jianbo Sun and Uttiya Basu

18. il-10 gene locus DNA methylation in regulatory B cells Silvia Tonon, Eleonora Martinis, Carlo E.M. Pucillo and Francesca Mion

19. B cell commitment to IL-10 production: the Vert-X Il10egfp mouse Akihiko Oka, Bo Liu, Jeremy W. Herzog and R. Balfour Sartor

Part V: Study of regulatory B cells in pathological settings
20. Regulatory B cells in experimental mouse models of arthritis Diana E. Matei, Claudia Mauri and Elizabeth C. Rosser

21. Regulatory and IgE+ B cells in allergic asthma John Paul Oliveria, Rita Agayby and Gail M. Gauvreau

22. Regulatory B cells in type 1 diabetes Joanne Boldison, Larissa Camargo Da Rosa and F. Susan Wong

23. Analysis of regulatory B cells in experimental autoimmune uveitis Jin Kyeong Choi and Charles E. Egwuagu

24. Purification and immunophenotypic characterization of human CD24hiCD38hi and CD24hiCD27+ regulatory B cells in transplantation Rowa. Y. Alhabbab and Giovanna Lombardi