Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.603958
Title: A study of soluble MHC class I in experimental transplantation
Author: Henderson, M. L.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2005
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Abstract:
In this study, soluble MHC class I (sClassI) levels were investigated in rat models of allogenic heart and liver transplantation. This necessitated the synthesis and refolding of two allelic forms (RT1A.a and RT1A.1) of recombinant sClassI (rsClass I) molecules and the development of sensitive ELISAs, using the rsClass I as standards, in order to quantify the levels of circulating sClass I in the serum of rat transplant recipients. Two components of sClass I, the heavy chain (lacking the transmembrane and cytoplasmic domain) and the beta2-microglobulin (β2m), were prepared using a bacterial expression system. Acceptable yields of RT1A.a heavy chain were routinely achieved but production of RT1A.1 required modification of the DNA sequence. High expression of RT1A.1 heavy chain was only achieved following the introduction of silent mutations to the sequence, which were believed to have altered the mRNA secondary structure leading to improved transcription. Each allelic heavy chain protein was refolded with βb2m nd peptide using a dilution method. The products of the refold were purified by gel filtration and FPLC. Allele-specific ELISAs were used to measure the levels of donor and recipient sClass I serum taken at different time points from Lewis recipients of DA heterotopic cardiac transplants (HCT) or DA liver transplants (OLT). Following HCT, DA donor strain sClass I was not detectable in the serum of Lewis recipients whereas recipient strain sClass I levels rose steadily. In contrast, following OLT, the levels of DA donor sClass I reached values comparable to normal DA rats and remained at this level for the duration of the experiment. Interestingly, following OLT and removal of the recipient’s own liver, the levels of recipient sClass I also rose. The results of this study are discussed with reference to the current opinion on the source and relevance of sClass I in transplantation.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.603958  DOI: Not available
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