Abstract
Langerhans cells (LCs) are antigen-presenting cells responsible for initiating an immune response against human papillomaviruses (HPVs) entering the epithelial layer in vivo as they are the first immune cell that HPV comes into contact with. LCs become activated in response to foreign antigens, which causes internal signaling resulting in the increased expression of co-stimulatory molecules and the secretion of inflammatory cytokines. Functionally activated LCs are then capable of migrating to the lymph nodes where they interact with antigen-specific T cells and initiate an adaptive T-cell response in vivo. However, HPV has evolved in a manner that suppresses LC function, and thus the induction of antigen-specific T cells is hindered. While many methods exist to monitor the activity of LCs in vitro, the migration and induction of cytotoxic T cells is ultimately indicative of a functional immune response. Here, methods in analyzing functional migration and induction of antigen-specific T cells after stimulation of LCs with HPV virus-like particles in vitro are described.
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References
Walboomers JMM et al (1999) Human papillomavirus is a necessary cause of invasive cervical cancer worldwide. J Pathol 189(1):12–19
Bosch F, Manos M, Munoz N, Sherman M, Jansen A, Peto J, Schiffman M, Moreno V, Kurman R, Shah K (1995) Prevalence of human papillomavirus in cervical cancer: a worldwide perspective. International Biological Study on Servical Cancer (IBSCC) Study Group. J Natl Cancer Inst 87:796–802
Stanley MA, Pett MR, Coleman N (2007) HPV: from infection to cancer. Biochem Soc Trans 35(6):1456–1460
Merad M, Ginhoux F, Collin M (2008) Origin, homeostasis and function of Langerhans cells and other langerin-expressing dendritic cells. Nat Rev Immunol 8(12):935–947
Cumming SA, Cheun-Im T, Milligan SG, Graham SV (2008) Human Papillomavirus type 16 late gene expression is regulated by cellular RNA processing factors in response to epithelial differentiation. Biochem Soc Trans 36:522–524
Kirnbauer R et al (1992) Papillomavirus L1 major capsid protein self-assembles into virus-like particles that are highly immunogenic. Proc Natl Acad Sci U S A 89(24):12180–12184
Buck CB et al (2008) Arrangement of L2 within the papillomavirus capsid. J Virol 82(11):5190–5197
Fahey LM et al (2009) A major role for the minor capsid protein of human papillomavirus type 16 in immune escape. J Immunol 183(10):6151–6156
Yan M et al (2004) Despite differences between dendritic cells and Langerhans cells in the mechanism of papillomavirus-like particle antigen uptake, both cells cross-prime T cells. Virology 324(2):297–310
Fausch SC et al (2002) Human papillomavirus virus-like particles do not activate Langerhans cells: a possible immune escape mechanism used by human papillomaviruses. J Immunol 169(6):3242–3249
Fausch SC, Da Silva DM, Kast WM (2003) Differential uptake and cross-presentation of human papillomavirus virus-like particles by dendritic cells and Langerhans cells. Cancer Res 63:3478–3482
Fausch SC et al (2005) Human papillomavirus can escape immune recognition through Langerhans cell phosphoinositide 3-kinase activation. J Immunol 174(11):7172–7178
Peiser M et al (2008) Human Langerhans cells selectively activated via Toll-like receptor 2 agonists acquire migratory and CD4 + T cell stimulatory capacity. J Leukoc Biol 83(5):1118–1127
Larsen CP et al (1990) Migration and maturation of Langerhans cells in skin transplants and explants. J Exp Med 172(5):1483–1493
Albert ML, Sauter B, Bhardwaj N (1998) Dendritic cells acquire antigen from apoptotic cells and induce class I-restricted CTLs. Nature 392(6671):86–89
Banchereau J, Steinman RM (1998) Dendritic cells and the control of immunity. Nature 392(6673):245–252
Acknowledgements
WM Kast holds the Walter A. Richter Cancer Research Chair. The methods in this chapter were developed with support from the Karl. H and Ruth M. Balz Trust, Sammie’s circle and NIH Grant R01 CA74397 in the Immune Monitoring Core Facility of the Norris Comprehensive Cancer Center that is supported by NIH Grant 5P30 CA014089 from the NCI. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NCI or the NIH.
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Yan, L., Woodham, A.W., Da Silva, D.M., Kast, W.M. (2015). Functional Analysis of HPV-Like Particle-Activated Langerhans Cells In Vitro. In: Keppler, D., Lin, A. (eds) Cervical Cancer. Methods in Molecular Biology, vol 1249. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2013-6_25
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DOI: https://doi.org/10.1007/978-1-4939-2013-6_25
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-2013-6
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