Porcine CFSE mixed lymphocyte reaction and PKH-26 cell-mediated lympholysis assays

Transplant Immunology
Manei OkuKazuhiko Yamada

Abstract

Mixed lymphocyte reaction (MLR) and cell-mediated lympholysis (CML) are widely used to assess T cell responses. A major limitation of the traditional MLR and CML assays is that they require radioisotope labeling with (3)H for MLR and (51)Cr for CML, thereby limiting their use to laboratories with the capabilities to deal safely with these materials. Recently, flow cytometry with CFSE labeling has been used to detect cell division in rodent and human assays, and flow cytometry with PKH-26 labeling has been used to study cytotoxicity in murine models. Partially inbred miniature swine provide a unique large animal preclinical model for experimental transplantation, helping to bridge the gap between rodent and clinical studies. In this study, we modified the reported CFSE and PKH-26 labeling procedures for use with porcine cells, and established that these radioactive-free MLR and CML assays are comparable to traditional radioactive CML and MLR assays for assessing immunologic responses in miniature swine. To our knowledge, this is the first report that has directly compared the traditional CML/MLR with radiation-free CML/MLR in MHC-defined swine models. The aim of this study is to establish non-radiolabeled CSFE and PKH-26 labelin...Continue Reading

References

Mar 1, 1977·Transplantation Proceedings·G S LeightS A Rosenberg
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Sep 6, 2003·Methods : a Companion to Methods in Enzymology·Karin Fischer, Andreas Mackensen
Jan 25, 2006·Transplant Immunology·Kenneth J WoodsideJohn A Daller
Dec 7, 2006·Proceedings of the National Academy of Sciences of the United States of America·Shuji NoboriKazuhiko Yamada
Jan 25, 2008·The New England Journal of Medicine·Tatsuo KawaiDavid H Sachs
Jan 25, 2008·The New England Journal of Medicine·John D ScandlingSamuel Strober
Jan 25, 2008·The New England Journal of Medicine·Stephen I AlexanderMichael O Stormon

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Citations

Aug 27, 2010·Veterinary Immunology and Immunopathology·T S DalgaardH R Juul-Madsen
Dec 17, 2014·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Jinyao ChenLishi Zhang

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