Abstract
Bronchus-associated lymphoid tissue (BALT) forms spontaneously in the lung after pulmonary infection and has been identified as a highly organized lymphoid structure supporting the efficient priming of T cells in the lung. To explore the mechanisms and instructive signals controlling BALT neogenesis we used both, a single dose of vaccinia virus MVA and repeated inhalations of heat-inactivated Pseudomonas aeruginosa (P. aeruginosa). Intranasal administration of both pathogens induces highly organized BALT but distinct pathways and molecules are used to promote the development of BALT. Here, we describe the induction and phenotype of the distinct types of BALT as well as the immunofluorescence microscopy-based analysis of the induced lymphoid tissue in the lung.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Neyt K, Perros F, GeurtsvanKessel CH et al (2012) Tertiary lymphoid organs in infection and autoimmunity. Trends Immunol 33:297–305
Xu B, Wagner N, Pham LN et al (2003) Lymphocyte homing to bronchus-associated lymphoid tissue (BALT) is mediated by L-selectin/PNAd, alpha4beta1 integrin/VCAM-1, and LFA-1 adhesion pathways. J Exp Med 197:1255–1267
Moyron-Quiroz JE, Rangel-Moreno J, Kusser K et al (2004) Role of inducible bronchus associated lymphoid tissue (iBALT) in respiratory immunity. Nat Med 10:927–934
Sminia T, van der Brugge-Gamelkoorn GJ, Jeurissen SH (1989) Structure and function of bronchus-associated lymphoid tissue (BALT). Crit Rev Immunol 9:119–150
Halle S, Dujardin HC, Bakocevic N et al (2009) Induced bronchus-associated lymphoid tissue serves as a general priming site for T cells and is maintained by dendritic cells. J Exp Med 206:2593–2601
Kocks JR, Adler H, Danzer H et al (2009) Chemokine receptor CCR7 contributes to a rapid and efficient clearance of lytic murine gamma-herpes virus 68 from the lung, whereas bronchus-associated lymphoid tissue harbors virus during latency. J Immunol 182:6861–6869
Toyoshima M, Chida K, Sato A (2000) Antigen uptake and subsequent cell kinetics in bronchus-associated lymphoid tissue. Respirology 5:141–145
Rangel-Moreno J, Carragher DM, de la Luz Garcia-Hernandez M et al (2011) The development of inducible bronchus-associated lymphoid tissue depends on IL-17. Nat Immunol 12:639–646
Meyer H, Sutter G, Mayr A (1991) Mapping of deletions in the genome of the highly attenuated vaccinia virus MVA and their influence on virulence. J Gen Virol 72(Pt 5):1031–1038
Kremer M, Volz A, Kreijtz JHCM et al (2012) Easy and efficient protocols for working with recombinant vaccinia virus MVA. Methods Mol Biol 890:59–92
Liberati NT, Urbach JM, Miyata S et al (2006) An ordered, nonredundant library of Pseudomonas aeruginosa strain PA14 transposon insertion mutants. Proc Natl Acad Sci 103:2833–2838
Fleige H, Ravens S, Moschovakis GL et al (2014) IL-17-induced CXCL12 recruits B cells and induces follicle formation in BALT in the absence of differentiated FDCs. J Exp Med 211:643–651
Foo SY, Zhang V, Lalwani A et al (2015) Regulatory T cells prevent inducible BALT formation by dampening neutrophilic inflammation. J Immunol 194:4567–4576
Kocks JR, Davalos-Misslitz ACM, Hintzen G et al (2007) Regulatory T cells interfere with the development of bronchus-associated lymphoid tissue. J Exp Med 204:723–734
Acknowledgment
We would like to thank Stephan Halle and Tim Worbs for valuable advices in establishing the infection model, Susanne Häußler and Gerd Sutter for providing P. aeruginosa and MVA respectively. This work was supported by Deutsche Forschungsgemeinschaft (DFG) grants SFB587-B3 and SFB900-B1 and by ERC grant 322645 LYMPHATICS HOMING to R. Förster.
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Fleige, H., Förster, R. (2017). Induction and Analysis of Bronchus-Associated Lymphoid Tissue. In: Clausen, B., Laman, J. (eds) Inflammation. Methods in Molecular Biology, vol 1559. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6786-5_13
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6786-5_13
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6784-1
Online ISBN: 978-1-4939-6786-5
eBook Packages: Springer Protocols