Abstract
Cellular studies are essential in the xenotransplantation field in order to investigate the cellular immune responses triggered by xenogeneic cells and identify the key molecules involved. A series of functional studies can be conducted with this purpose that include treatment with proinflammatory cytokines and xenogeneic cell-based assays that put together pig cells and human leukocytes such as monocytes, NK cells, and T cells. The choice of the pig cell type is critical to appropriately model the transplant setting of interest. Thus, pig endothelial cells are commonly used for studying the rejection process of vascularized organs. Treatment with cytokines allows studying the regulation of adhesion, costimulatory molecules, and receptors involved in triggering the immune response in an attempt to reproduce the more complex in vivo situation. The adhesion assays are used to determine the capacity of human leukocytes to adhere to porcine cells under various conditions. Furthermore, we describe coculture, costimulatory, and cytotoxicity assays for investigating the cellular and molecular mechanisms that take place during the xenogeneic immune response.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ashton-Chess J, Roussel JC, Manez R et al (2003) Cellular participation in delayed xenograft rejection of hCD55 transgenic pig hearts by baboons. Xenotransplantation 10:446–453
Davila E, Byrne GW, LaBreche PT et al (2006) T-cell responses during pig-to-primate xenotransplantation. Xenotransplantation 13:31–40
Tseng YL, Kuwaki K, Dor FJ et al (2005) alpha1,3-Galactosyltransferase gene-knockout pig heart transplantation in baboons with survival approaching 6 months. Transplantation 80:1493–1500
Cardona K, Korbutt GS, Milas Z et al (2006) Long-term survival of neonatal porcine islets in nonhuman primates by targeting costimulation pathways. Nat Med 12:304–306
Hering BJ, Wijkstrom M, Graham ML et al (2006) Prolonged diabetes reversal after intraportal xenotransplantation of wild-type porcine islets in immunosuppressed nonhuman primates. Nat Med 12:301–303
Murray AG, Khodadoust MM, Pober JS, Bothwell AL (1994) Porcine aortic endothelial cells activate human T cells: direct presentation of MHC antigens and costimulation by ligands for human CD2 and CD28. Immunity 1:57–63
Rollins SA, Kennedy SP, Chodera AJ et al (1994) Evidence that activation of human T cells by porcine endothelium involves direct recognition of porcine SLA and costimulation by porcine ligands for LFA-1 and CD2. Transplantation 57:1709–1716
Costa C, Barber DF, Fodor WL (2002) Human NK cell-mediated cytotoxicity triggered by CD86 and Galα1,3-Gal is inhibited in genetically modified porcine cells. J Immunol 168:3808–3816
Sullivan JA, Oettinger HF, Sachs DH, Edge AS (1997) Analysis of polymorphism in porcine MHC class I genes: alterations in signals recognized by human cytotoxic lymphocytes. J Immunol 159:2318–2326
Seebach JD, Comrack C, Germana S, LeGuern C, Sachs DH, DerSimonian H (1997) HLA-Cw3 expression on porcine endothelial cells protects against xenogeneic cytotoxicity mediated by a subset of human NK cells. J Immunol 159:3655–3661
Forte P, Lilienfeld BG, Baumann BC, Seebach JD (2005) Human NK cytotoxicity against porcine cells is triggered by NKp44 and NKG2D. J Immunol 175:5463–5470
Li S, Waer M, Billiau AD (2009) Xenotransplantation: role of natural immunity. Transpl Immunol 21:70–74
Jin R, Greenwald A, Peterson MD, Waddell TK (2006) Human monocytes recognize porcine endothelium via the interaction of galectin 3 and α-GAL. J Immunol 177:1289–1295
Hauzenberger E, Hauzenberger D, Hultenby K, Holgersson J (2000) Porcine endothelium supports transendothelial migration of human leukocyte subpopulations: anti-porcine vascular cell adhesion molecule antibodies as species-specific blockers of transendothelial monocyte and natural killer cell migration. Transplantation 69:1837–1849
Ide K, Wang H, Tahara H et al (2007) Role for CD47-SIRPα signaling in xenograft rejection by macrophages. Proc Natl Acad Sci USA 104:5062–5066
Sommaggio R, Máñez R, Costa C (2009) TNF, pig CD86 and VCAM-1 identified as potential targets for intervention in xenotransplantation of pig chondrocytes. Cell Transplant 18:1381–1393
Sultan P, Murray AG, McNiff JM et al (1997) Pig but not human interferon-gamma initiates human cell-mediated rejection of pig tissue in vivo. Proc Natl Acad Sci USA 94:8767–8772
Acknowledgments
This work was supported by Ministerio de Educación y Ciencia (SAF2008-00499) and a grant from Fundación de Investigación Médica Mútua Madrileña (338/05) to C.C. R.S. was supported by a 1-year fellowship from Fundació catalana de trasplantament and M.P.-C. by an IDIBELL fellowship.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Sommaggio, R., Pérez-Cruz, M., Costa, C. (2012). Cellular Studies for In Vitro Modeling of Xenogeneic Immune Responses. In: Costa, C., Máñez, R. (eds) Xenotransplantation. Methods in Molecular Biology, vol 885. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-845-0_7
Download citation
DOI: https://doi.org/10.1007/978-1-61779-845-0_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-844-3
Online ISBN: 978-1-61779-845-0
eBook Packages: Springer Protocols