Abstract
The generation of class-switched, high-affinity, antibody-producing B cells plays a critical role in the establishment of type 2 immunity to intestinal helminths as well as in the pathogenesis of allergy and asthma. The generation of these high-affinity, antibody-producing B cells occurs in germinal centers (GC) and relies on interactions with follicular dendritic cells (FDCs) and T follicular helper (Tfh) cells. One critical mediator produced by Tfh cells in GCs is interleukin-4 (IL-4). Tfh-derived IL-4 drives class switching to type 2 antibody isotypes IgE and IgG1 and is required for high-affinity IgG1 production. In vivo detection of IL-4-expressing Tfh cells is required to better understand the role of these cells during the GC response. Detection of IL-4-expressing cells has been greatly improved by the generation of the IL-44get reporter mice, which read out IL-4 expression as green fluorescent protein (GFP). Much has been learned from these mice with regard to type 2 immunity using flow cytometry and immunohistochemistry. However, these methods do not allow the study of cellular behavior and interactions in real time. In contrast, multi-photon microscopy allows for deep tissue imaging and tracking of multiple cell types in intact tissues over time. Here, we describe a protocol for in vivo detection of IL-4-expressing Tfh cells in an explanted popliteal lymph node by multi-photon microscopy. The dynamics of Tfh cell motility and their interactions with FDC networks in the GCs were analyzed.
The original version of this chapter was revised. A correction to this chapter can be found at https://doi.org/10.1007/978-1-4939-7896-0_31
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Change history
05 September 2018
Correction to: R. Lee Reinhardt (ed.), Type 2 Immunity: Methods and Protocols, Methods in Molecular Biology, vol. 1799, https://doi.org/10.1007/978-1-4939-7896-0
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Acknowledgments
This research was funded in part by NIH grants R01AI125553 (J.J.), R01DK111733 (R.S.F), and R01AI119004 (R.L.R). The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
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Dell’Aringa, M., Reinhardt, R.L., Friedman, R.S., Jacobelli, J. (2018). Live Imaging of IL-4-Expressing T Follicular Helper Cells in Explanted Lymph Nodes. In: Reinhardt, R. (eds) Type 2 Immunity. Methods in Molecular Biology, vol 1799. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-7896-0_17
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DOI: https://doi.org/10.1007/978-1-4939-7896-0_17
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