Analysis of Draining Leukocytes from the Abdominal Cavity Monitors Immune Events after Intestinal Transplant

DOMINIK MEIER; HERNÁN CAGNOLA; RAMISCH DIEGO; CAROLINA RUMBO; GUILLERMO DOCENA; FERNANDO CHIRDO; GABRIEL GONDOLES; MARTIN RUMBO

Congreso; 1st. French-Argentinean Congress of Immunology, November 2-5, 2010. Buenos Aires, Argentina.; 2010

<!-- /* Font Definitions */ @font-face {font-family:"Arial Narrow"; panose-1:2 11 5 6 2 2 2 3 2 4; mso-font-charset:0; mso-generic-font-family:swiss; mso-font-pitch:variable; mso-font-signature:647 0 0 0 159 0;} /* Style Definitions */ p.MsoNormal, li.MsoNormal, div.MsoNormal {mso-style-parent:""; margin:0cm; margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Times New Roman"; mso-fareast-font-family:"Times New Roman";} @page Section1 {size:612.0pt 792.0pt; margin:70.85pt 3.0cm 70.85pt 3.0cm; mso-header-margin:36.0pt; mso-footer-margin:36.0pt; mso-paper-source:0;} div.Section1 {page:Section1;} --> During intestinal transplant (ITx) operation, intestinal lymphatics are not reconstituted. Consequently, migrating immune cells drain freely into the abdominal cavity and could be recovered though a drainage of the intestinal cavity used for clinical postoperative surveillance. Our aim was to evaluate whether leukocytes migrating from the transplanted intestine could be recovered from the abdominal draining fluid and to determine potential applications of the assessment of draining cellular populations to basic and clinical studies. Draining fluid was collected by a peritoneal drainage system in the early postoperative period of 7 ITx patients. Cell composition of the abdominal draining fluid was analyzed by flow cytometry during the first 14 post-ITx days. Cell sorting and molecular fingerprinting by short tandem repeat amplification was performed. The correlation between analyzed parameters and clinical evolution was evaluated. The main populations in the draining fluid were CD3+CD4+CD8-, CD3+CD8+CD4- and CD3-HLA-DR+CD19+ lymphocytes, although several minor populations were identified. Cellular pattern varies along the post-ITx period in non-complicated recipients from a mixed leukocyte pattern to an exclusively lymphocytic pattern. We could associate changes in draining cell patterns to early 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. This study demonstrates that 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. Furthermore, this is a unique opportunity to study different immune cell populations migrating from the mucosal intestinal site..