T-Follicular Helper Cells Methods and Protocols

The volume provides a comprehensive overview of methods that have been developed for the study of Tfh and Tfr biology in mice, and in different human diseases. The chapters detail methods to investigate the function of Tfh cells in vivo and in vitro, including the use of mathematical models to study GC dynamics. 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-Follicular Helper Cells: Methods and Protocols aims to be a useful practical guide to researchers to help further their study in this field.

 Part I: Identification of Tfh and Tfr cells in humans and mice

1.         Complete Visualization of T Follicular Helper Cells in Germinal Centers

Liat Stoler-Barak and Ziv Shulman

2.         In vivo imaging of Tfh cells  

Sandra Nakandakari-Higa and Johanne T. Jacobsen

3.         Circulating T Follicular Helper Subsets in Human Blood

Yayoi Iwamoto and Hideki Ueno

4.         Identification of Human Circulating and Tissue T Follicular Regulatory (Tfr) Cells by Flow Cytometry

Filipa Ribeiro, Pedro Ávila, João Eurico Fonseca, and Luis Graca

5.         Assessment of circulating Tfh and Tfr by mass cytometry

James B. Wing and Shimon Sakaguchi

6.         Identification of T peripheral helper (Tph) cells

Vanessa S. Wacleche, Runci Wang, and Deepak A. Rao

7.         Identification of non-classical follicular T cells 

Alan Courey-Ghaouzi and Mauro Gaya

8.         Identification of follicular T cells in the gut

Hristo Georgiev, Georgia Papadogianni, and Günter Bernhardt

Part II: Development and function of Tfh and Tfr cells

9.         Intravital imaging of Tfh dynamics

Bo Liu and Hai Qi

10.     A method to analyze models of the dynamics of GCs

J. Faro and E. Faro

11.     Identification of Antigen Presenting Cell subsets supporting human Tfh differentiation Lamine Alaoui, Melanie Durand, and Elodie Segura

12.     The use of peptide-MHCII tetramers to identify antigen-specific T follicular helper and T follicular regulatory cells

Meryem Aloulou and Nicolas Fazilleau

13.     Assessment of the TCR repertoire of human circulating T follicular helper cells Mengyun Hu and Federica Sallusto

14.     Assessing in vivo T cell-dependent antigen-specific antibody responses

Markus M. Xie and Alexander L. Dent

15.     Tfh-mediated and Tfr-suppressed antigen driven IgG and IgE assays

Rachel L. Clement, Benjamin L. Hanson, and Peter T. Sage

Part III: Tfh and Tfr cells in infectious and inflammatory diseases

16.     Assessment of the impact of cytokines on TFH, TREG, and TFR cell populations after influenza infection

Amber Papillion, Meagan M. Jenkins, and André Ballesteros-Tato

17.     Evaluation of human circulating T follicular helper cells in influenza- and SARS-CoV-2-specific B cell immunity

Marios Koutsakos, Katherine Kedzierska, and Thi H. O. Nguyen

Sulan Yu, Meiling Wu, Yun Feng, Jiangang Shen, Liwei Lu, and Xiang Lin

19.     Identification of Human T Follicular Cells in Ectopic Lymphoid Structures

Rodrigo Pedroso, Filipa Ribeiro, Ana Rita Pires, Luis Graca, and Válter R. Fonseca

20.     Understanding T Follicular helper cells in asthma through murine models of allergic airway disease 

Faith I. Uwadiae and James A. Harker

21.     Therapeutic modulation of T follicular helper cells by low-dose IL-2 treatment

Zhian Chen, Kaili Liang, Jun Deng, and Di Yu

22.     3D tissue explant and single-cell culture systems for ex vivo drug testing on Tfh cells derived from human adenoid tonsils

Angelika Schmidt and Dirk Baumjohann