The aim of this project was to develop an in vitro functional assay for the prediction of
allograft rejection following renal transplantation. This assay was also used to study
acute GVHD following identical sibling bone marrow transplantation. Lymphocyte
cytokine profiles were measured by ELISA (protein secretion) and flow cytometry
(cytokine expression) following mitogen stimulation and MLR. In normal individuals,
considerable inter-individual variations were found in both protein secretion (IL-2, IL-
4, IL-10 and IFN-γ) and cytokine expression (IL-2 and IFN-γ). Strong relationships
were found between IL-2 protein and expression, IL-2 and IL-10 protein, and IL-10
and IFN-γ protein secretion. Analysis of cytokine gene polymorphisms showed no
correlation between IFN-y protein secretion, frequency or gene polymorphism.
Pre-transplant MLRs were set up between renal transplant patient/donors pairs and
cytokine protein secretion (IL-2, IL-4, IL-6, IL-10 and IFN-γ) measured by ELISA.
Analysis was performed to ascertain predictive factors of allograft rejection. Inter-individual
variations were found for all cytokine profiles. Significant correlations were
found between individual cytokine protein profiles including IL-10 and IFN-γ. In
addition, a correlation was found between HLA-DR mismatching and both IL-10 and
IFN-γ protein secreted in the MLR. Primary univariate analysis revealed that HLA and
HLA-DR mismatching, female donor sex, MLR-stimulated IL-10, MLR-stimulated
IFN-γ and spontaneous IL-4 secretion were associated with an increased risk of
rejection. Multivariate analysis showed the strongest correlations for predicting risk of
rejection to be female donor sex, HLA mismatching and MLR-stimulated IL-l 0
secretion. A combination of high HLA mismatching and high IL-l 0 secretion in MLR
gave the highest risk of rejection (RR=25.5). Finally, cytokine secretion decreased
when measured post-transplant. Prediction of graft survival could not be analysed due
to the low number (n=6) of patients that suffered graft failure in the group.
Cytokine protein secretion (IL-2, IL-4, IL-10 and IFN-γ) in MLR was also studied for
prediction of GVHD after bone marrow transplantation. There was a very low MLR response by all BMT pairs, therefore analysis could not be performed.
Date of Award | 1999 |
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Original language | English |
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Awarding Institution | |
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CYTOKINE PROFILES AND THEIR RELEVANCE TO HUMAN TRANSPLANTATION
CARTWRIGHT, N. H. (Author). 1999
Student thesis: PhD