Analysis of Draining Leukocytes from the Abdominal Fluid for monitoring Immune Events after Intestinal Transplant

DOMINIK MEIER; HERNÁN CAGNOLA; DIEGO RAMISCH; CAROLINA RUMBO; FERNANDO CHIRDO; GUILLERMO DOCENA; GABRIEL E. GONDOLES; MARTÍN RUMBO

Bologna, Italy.

Congreso; XI International Small Bowel Transplant Symposium.; 2009

International Society of Intestinal Transplantation

Introduction (Max 50 words): During intestinal transplant (ITx) operation, intestinal lymphatics are not reconstituted. Our aim was to evaluate whether leukocytes migrating from the transplanted intestine could be recovered from the abdominal draining fluid and to determine the potential applications of the assessment of draining cellular populations to basic and clinical studies. Methods (Max 50 words): Six consecutive ITx patients were included; the cell composition of the abdominal draining fluid was analyzed the first 15 post-ITx days by flow cytometry. Cell sorting of CD8+ T cells and molecular fingerprinting by short tandem repeat amplification was performed. The correlation between analyzed parameters and clinical evolution was evaluated. Results (Max 100 words): The main populations in the draining fluid were CD3+CD4+CD8-, CD3+CD8+CD4- and CD3-HLA-DR+CD19+ lymphocytes. Cellular pattern varies along the post-ITx period in non-complicated recipients from a mixed leukocyte pattern to an exclusively lymphocytic pattern (Fig1, a-b). We could associate changes in draining cell patterns to early rejection (Fig 1,c)  or infections (Fig1,d). Graft derived lymphocytes were recognized by genetic fingerprinting of CD8+ sorted T cells. At days 1-2, donor T cells were detected in the draining fluid (50% of total CD8+ cells) and were mostly replaced by day 4 after ITx (<5%), confirming that cells migrating from the graft can be recovered in the draining fluid. Conclusions (Max 50 words): This study demonstrates that the cell analysis of the draining fluid from ITx recipients may provide a useful approach for monitoring changes leukocyte populations migrating from the graft during the early post-transplant period. This analysis might be useful for clinical management and also to gain insight on intestinal transplant immunobiology.