Tracking Early T Follicular Helper Cell Differentiation In Vivo

  • Dirk Baumjohann
  • K. Mark Ansel
Part of the Methods in Molecular Biology book series (MIMB, volume 1291)


T follicular helper (Tfh) cells provide essential help to B cells for the generation of high-affinity antibodies. These mechanisms provide the basis for the success of modern vaccines, but dysregulated Tfh cell responses are also linked to autoimmune diseases. In addition to their established role in driving humoral immunity, Tfh cells are gaining attention for their role in other processes of the adaptive immune system. For example, Tfh cells may serve as transitional differentiation intermediates during effector and memory T-helper cell differentiation and as a reservoir of HIV-infected cells. While B cells are required for the full maturation and maintenance of Tfh cell responses, they are dispensable for the initial induction of the Tfh cell phenotype, which occurs at the priming stage through interaction with dendritic cells. Nevertheless, the precise mechanisms of these early events during Tfh cell differentiation remain relatively unknown. Here, we describe a method for tracking early Tfh cell differentiation by following cell division kinetics and phenotypic changes of recently activated antigen-specific CD4+ T cells in vivo. As an example, we use this method to visualize the requirements for T cell-expressed CD28 for the differentiation of CXCR5+Bcl6+ Tfh cells.

Key words

T follicular helper cells Follicular helper T cells Tfh cells Flow cytometry FACS CFSE CellTrace Violet CTV T-dependent antibody response Bcl6 Bcl-6 CXCR5 



This work was supported by the National Multiple Sclerosis Society (D.B.), the UCSF Program for Breakthrough Biomedical Research, funded in part by the Sandler Foundation (D.B.), and the NIH (P01 HL107202, R01 HL109102).


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Microbiology & Immunology, Sandler Asthma Basic Research CenterUniversity of California San FranciscoSan FranciscoUSA
  2. 2.Institute for ImmunologyLudwig-Maximilians-UniversityMunichGermany

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