Abstract
Humoral immune responses depend on B cells encountering antigen (Ag) in lymph nodes (LNs) draining infection sites, getting activated, interacting with different cells, proliferating and differentiating into antibody (Ab)-secreting cells. Each of these events occurs in distinct LN sub-compartments, requiring the migration of B cells from niche to niche in a fast and tightly coordinated fashion. While some of the rules that characterize B cell behavior in secondary lymphoid organs have been elucidated at the population level, we have only limited knowledge of the precise dynamics of B cell interactions with different kinds of LN cells at the single-cell level. Here, we describe in detail an intravital microscopy technique that allows the analysis of B cell dynamic behavior in the popliteal lymph node of anesthetized mice at high spatial and temporal resolution. A detailed understanding of the spatiotemporal dynamics of B cells within secondary lymphoid organs may lead to novel, rational vaccine strategies aimed at inducing rapid and long-lived humoral immune responses.
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References
Pereira JP, Kelly LM, Cyster JG (2010) Finding the right niche: B-cell migration in the early phases of T-dependent antibody responses. Int Immunol 22:413–419. https://doi.org/10.1093/intimm/dxq047
Andrian v UH, Mempel TR (2003) Homing and cellular traffic in lymph nodes. Nat Rev Immunol 3:867–878. https://doi.org/10.1038/nri1222
Halin C, Mora JR, Sumen C, Andrian v UH (2005) In vivo imaging of lymphocyte trafficking. Annu Rev Cell Dev Biol 21:581–603. https://doi.org/10.1146/annurev.cellbio.21.122303.133159
Bajénoff M, Egen JG, Qi H et al (2007) Highways, byways and breadcrumbs: directing lymphocyte traffic in the lymph node. Trends Immunol 28:346–352. https://doi.org/10.1016/j.it.2007.06.005
Batista FD, Harwood NE (2009) The who, how and where of antigen presentation to B cells. Nat Rev Immunol 9:15–27. https://doi.org/10.1038/nri2454
Germain RN, Robey EA, Cahalan MD (2012) A decade of imaging cellular motility and interaction dynamics in the immune system. Science 336:1676–1681. https://doi.org/10.1126/science.1221063
Cyster JG (2010) B cell follicles and antigen encounters of the third kind. Nat Immunol 11:989–996. https://doi.org/10.1038/ni.1946
Kuka M, Iannacone M (2014) The role of lymph node sinus macrophages in host defense. Ann N Y Acad Sci 1319:38–46. https://doi.org/10.1111/nyas.12387
Gonzalez SF, Degn SE, Pitcher LA et al (2011) Trafficking of B cell antigen in lymph nodes. Annu Rev Immunol 29:215–233. https://doi.org/10.1146/annurev-immunol-031210-101255
Victora GD, Mesin L (2014) Clonal and cellular dynamics in germinal centers. Curr Opin Immunol 28C:90–96. https://doi.org/10.1016/j.coi.2014.02.010
Mempel TR, Henrickson SE, Andrian v UH (2004) T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases. Nature 427:154–159. https://doi.org/10.1038/nature02238
Murooka TT, Mempel TR (2012) Multiphoton intravital microscopy to study lymphocyte motility in lymph nodes. Methods Mol Biol 757:247–257. https://doi.org/10.1007/978-1-61779-166-6_16
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Sammicheli, S., Kuka, M., Iannacone, M. (2018). Intravital Imaging of B Cell Responses in Lymph Nodes. In: Ishii, M. (eds) Intravital Imaging of Dynamic Bone and Immune Systems . Methods in Molecular Biology, vol 1763. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7762-8_7
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DOI: https://doi.org/10.1007/978-1-4939-7762-8_7
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