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Triggering Positive Selection of Germinal Center B Cells by Antigen Targeting to DEC-205

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T follicular Helper Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1291))

Abstract

Germinal centers (GCs) are the site of maturation of antibody affinity and are thus of key importance to humoral immunity. The study of B-cell dynamics and selection within the GC has been hampered by the limited number of techniques available to manipulate GC output in vivo. Here, we describe an approach to trigger positive selection of B cells in vivo by targeting antigen specifically to a subpopulation of GC B cells via the surface lectin DEC-205 and forcing their interaction with T follicular helper cells. Targeted GC B cells can then be followed over time as they progress through the stages of positive selection.

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References

  1. Victora GD, Nussenzweig MC (2012) Germinal centers. Annu Rev Immunol 30:429–457

    Article  CAS  PubMed  Google Scholar 

  2. Victora GD et al (2010) Germinal center dynamics revealed by multiphoton microscopy with a photoactivatable fluorescent reporter. Cell 143:592–605

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  3. Clynes RA, Towers TL, Presta LG, Ravetch JV (2000) Inhibitory Fc receptors modulate in vivo cytotoxicity against tumor targets. Nat Med 6:443–446

    Article  CAS  PubMed  Google Scholar 

  4. Shih TA, Roederer M, Nussenzweig MC (2002) Role of antigen receptor affinity in T cell-independent antibody responses in vivo. Nat Immunol 3:399–406

    Article  CAS  PubMed  Google Scholar 

  5. Inaba K et al (1995) Tissue distribution of the DEC-205 protein that is detected by the monoclonal antibody NLDC-145. I. Expression on dendritic cells and other subsets of mouse leukocytes. Cell Immunol 163:148–156

    Article  CAS  PubMed  Google Scholar 

  6. Dudziak D et al (2007) Differential antigen processing by dendritic cell subsets in vivo. Science 315:107–111

    Article  CAS  PubMed  Google Scholar 

  7. Aida Y, Pabst MJ (1990) Removal of endotoxin from protein solutions by phase separation using Triton X-114. J Immunol Methods 132:191–195

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Gabriel D. Victora .

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© 2015 Springer Science+Business Media New York

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Pasqual, G., Angelini, A., Victora, G.D. (2015). Triggering Positive Selection of Germinal Center B Cells by Antigen Targeting to DEC-205. In: Espéli, M., Linterman, M. (eds) T follicular Helper Cells. Methods in Molecular Biology, vol 1291. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2498-1_10

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  • DOI: https://doi.org/10.1007/978-1-4939-2498-1_10

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2497-4

  • Online ISBN: 978-1-4939-2498-1

  • eBook Packages: Springer Protocols

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