Presented at:
British Society of Immunology
Harrogate, UK, December, 2005
Immune Monitoring in Renal Transplantation
Hernandez-Fuentes MP, Sagoo P, Jimenez E, Rovis F, Stevenson S, Warrens A, Lechler R
King's College London, United Kingdom; Imperial College London, United Kingdom
Prospective study of a transplant recipients' immune response to donor tissue would prove an invaluable resource if the likely outcome of transplant success could be predicted. We have adapted several routine laboratory techniques to study donor-specific responses generated via direct and indirect pathways of antigen presentation in renal transplant patients.
Quantitative assessment of donor-specific responses generated by the indirect pathway are made by stimulating recipient PBMCs with a preparation of solubilised donor membrane proteins in an IFNγ ELISpot assay. Frequencies of direct pathway donor-reactive CD8 and CD4 T cells are studied by stimulation with APCs purified from donor PBMCs or spleen, by IFNγ and IL-4 ELISpot. Direct T cell responder frequencies are also estimated by CFSE analysis. Depletion of CD4+CD25+ cells from responder populations in each assay further allows detection of Treg-mediated regulation of anti-donor responses.
Using these assays donor-specific hypo- and hyper-responsiveness is detected. This paper discusses molecular, cellular, technical and statistical basis of the immune monitoring assays developed, which provide an insight into the development of recipient responses and the governing role of Tregs.