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

MEIER D; CAGNOLA HH; RAMISCH D; RUMBO C; CHIRDO F; DOCENA G; GONDOLESI G; RUMBO M

Bologna

Congreso; XIth International Small Bowel Transplant Symposium; 2009

XIth International Small Bowel Transplant Symposium

Introduction: 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: Six consecutive ITx patients were included; the cell composition of the abdominal draining fluid was analyzed the first 14 post-ITx days by flow cytometry. Cell sorting and molecular fingerprinting by short tandem repeat amplification was performed. The correlation between analyzed parameters and clinical evolution was evaluated. Results: In spite of marked lymphopenia in peripheral blood, all patients analyzed showed a variable proportion of lymphocytes in abdominal draining fluid, that in all cases was of above the 50% of total cells present in this fluid. 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-c). We could associate changes in draining cell patterns to early events such as rejection or infections. 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 11 after ITx (<2%), confirming that cells migrating from the graft can be recovered in the draining fluid. Conclusions: This study demonstrates that the cell analysis of the draining fluid from ITx recipients may provide a useful approach for monitoring changes in graft immunobiology during the first 3 weeks post-transplant. This type of analysis might be serve to improve clinical management.