• Marie Weinreich Petersen
  • Anette Sand Svendsen
In order to improve transplantation outcome a new method for evaluation of crossmatch would be valuable. Aim: The present study had three aims. The first aim was to evaluate kidney transplantation outcome in relation to results from tests performed before transplantation. The second aim was to test the magnetic activated cell sorting (MACS) for separating T-and B-cells from isolated peripheral blood mononuclear cells (PBMC) and defrosted spleen. The third aim was to evaluate the Vi-CELL XR Cell Viability Analyzer (Vi-CELL) for analyzing lymphocyte concentration and viability. Material and methods: Clinical data was gathered from 147 recipients who had been kidney transplanted with deceased donor in the years 2011-2013 at Aarhus University Hospital, Skejby. MACS was used to separate T- and B-cells. Lymphocyte concentration and viability before and after separation were evaluated using flow cytometry. Samples with lymphocytes from defrosted spleen and PBMC were analyzed for lymphocyte concentration and viability with the Vi-CELL and flow cytometry, and the results were compared. Results: No significant results were obtained when evaluating different tests performed before transplantation and their relation to transplantation outcome (P > 0.05). However, the results indicated that presence of human leukocyte antigen-antibodies and donor specific antibodies in the recipient contributes to poor estimated glomerular filtration rate (eGFR) and higher risk of rejections. A difference in lymphocyte viability ≤ 5 PP between before and after separation with MACS was acceptable and seen for two out of three. However, it was not possible to reach an acceptable purification yield (> 90 %) and lymphocyte loss (< 10 %). It was not possible to set the Vi-CELL parameters to measure a lymphocyte concentration ≤ 15 % difference compared to flow cytometry. However, the difference in lymphocyte viability between the Vi-CELL and flow cytometer was acceptable for the majority of the samples (≤ 5 PP). Conclusion: It was found that kidney transplantation outcome could not be predicted by pre-transplantation tests. When testing the utility of MACS acceptable values were obtained for lymphocyte death, but not for lymphocyte loss and purification yield. Evaluation of the Vi-CELL for analyzing lymphocyte concentration and viability showed that the Vi-CELL was able to measure viability correct, but not lymphocyte concentration compared to flow cytometry.
Udgivelsesdato29 maj 2015
Antal sider60
Ekstern samarbejdspartnerAarhus University Hospital
Ledende overlæge, klinisk lektor Bjarne Kuno Møller bjmoel@rm.dk
ID: 213140713