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P782 A new compatibility test for donor selection for faecal microbiota transplantation in ulcerative colitis

Ponce-Alonso M.1, Garcia-Fernandez S.1, Aguilera L.2, Rodriguez-de-Santiago E.2, Foruny J.R.2, Roy G.3, DelCampo R.1, Canton R.1, Lopez-Sanroman A.*2

1Hospital Ramon y Cajal, Department of Microbiology, Madrid, Spain 2Hospital Ramon y Cajal, Department of Gastroenterology & Hepatology, Madrid, Spain 3Hospital Ramon y Cajal, Department of Immunology, Madrid, Spain

Background

Faecal Microbiota Transplantation (FMT) is an effective and safe treatment against Clostridium difficile infections (CDI). Its usefulness for the treatment of other conditions is being explored. A promising application is ulcerative colitis (UC) treatment, as patients' microbiota contributes to the bowel inflammation in this disease. However, FMT therapeutic success seems to be donor dependent in this setting. The aims of the present work were: 1) To design an individualized test to select the best faecal donor for each UC patient, and 2) To assess the post-FMT implantation of gut microbiota in a UC patient and to compare it with implantation ons CDI patient who used the same faecal donor.

Methods

A 40 year-old male with extensive moderate UC (E3S2) was approached and consented to be studied for protocol design. Disease was refractory to anti-TNF-α, anti-integrin, tacrolimus, and thiopurines. Patient was maintained on 15 mg/day of prednisone. Lymphoid cells from rectal biopsies were obtained by enzymatic digestion with collagenase and DNA-ase. Lymphocytes were faced against three gut microbiota samples from independent healthy donors to determine interleukin production in supernatants using the Cytometric Bead Array kit (B&D). Different incubation times (6, 18, 24 h) and microbiota concentrations (1, 1/100, 1/1000) were tested to select the optimal conditions. Donor faeces resulting in a milder inflammatory response were chosen for FMT. An unrelated CDI patient underwent also FMT with the same faeces, and was used as a positive control of faecal microbiota implantation, which was assessed by PCR-DGGE of the donor faeces, three UC faecal samples (basal, 15 days and 30 days after FMT) and a single CDI one (30 days after FMT).

Results

The optimal conditions of our test corresponded to a non-diluted faecal sample, combined with an 18 h incubation with lymphocytes. Markers that best discriminated the inflammatory response (higher response range) against the donor microbiota were IL-6 and TNF-α. Our protocol allowed the selection of a faecal donor for our UC patient, preventing the triggering of an inflammatory response in the intestinal immune system. FMT also allowed the reduction of prednisone doses in this patient as well as a clinical improvement of his symptoms. After FMT, PCR-DGGE gut microbiota fingerprints were indistinguishable between the healthy donor and both the UC and CDI patients, demonstrating an adequate microbiota implantation.

Conclusion

We propose this new test to select the most compatible gut microbiota donor for each UC-patient before the FMT. Although the effectiveness of this test should be validated in a higher number of UC-patients and donors, we obtained promising results in our single patient.