Uncomplicated diverticular disease: innate and adaptive immunity in human gut mucosa before and after rifaximin

Rossella Cianci, Simona Frosali, Danilo Pagliari, Paola Cesaro, Lucio Petruzziello, Fabio Casciano, Raffaele Landolfi, Guido Costamagna, Franco Pandolfi, Rossella Cianci, Simona Frosali, Danilo Pagliari, Paola Cesaro, Lucio Petruzziello, Fabio Casciano, Raffaele Landolfi, Guido Costamagna, Franco Pandolfi

Abstract

Background/aim: Uncomplicated diverticular disease (UDD) is a frequent condition in adults. The pathogenesis of symptoms remains unknown. Bacteria are able to interact with Toll-like receptors (TLRs) and to induce inflammation through both innate immunity and T-cell recruitment. We investigated the pattern of TLRs 2 and 4 and the intestinal homing in patients with UDD before and after a course of Rifaximin.

Methods: Forty consecutive patients with UDD and 20 healthy asymptomatic subjects were enrolled. Among UDD patients, 20 were assigned to a 2-month course of treatment with Rifaximin 1.2 g/day for 15 days/month and 20 received placebo. Blood sample and colonic biopsies were obtained from patients and controls. The samples were collected and analyzed at baseline and at the end of treatment. Flow cytometry was performed using monoclonal antibodies (CD3, CD4, CD8, CD103, TCR-gamma/delta, CD14, TLR2, and TLR4).

Results: In UDD, TLR2 and TLR4 expression on immune cell subpopulations from blood and mucosa of the affected colon are altered as compared with controls. Rifaximin treatment induced significant modifications of altered conditions.

Conclusions: Our data show the role of TLRs in the development of inflammation in UDD. TLRs distribution is altered in UDD and these alterations are reversed after antibiotic treatment. This trial is registered with ClinicalTrials.gov: NCT02068482.

Figures

Figure 1
Figure 1
Expression of TLRs in lymphocytes. (a, b) TLR2 and TLR4 in peripheral blood lymphocytes, at baseline and after Rifaximin and placebo treatments. (c, d) TLR2 and TLR4 in sigma mucosa lymphocytes at baseline and after Rifaximin and placebo treatments. (e) TLR2 and TLR4 in transverse mucosa lymphocytes at baseline and after Rifaximin treatment. The values reported in box-and-whiskers plots show the minimum, the 25th percentile, the median, the 75th percentile, and the maximum. In particular, the whiskers go down to the smallest value (minimum) and up to the largest one (maximum), the top and bottom of the box are the 25th and 75th percentiles, and the line in the middle of the box is the median corresponding to 50th percentile (***P < 0.001, **P < 0.01, *P < 0.05 patients versus control (health or baseline)); §§P < 0.01, §P < 0.05 between treatments. Healthy control (CTR), UDD patients (PZ), before Rifaximin (BR), after Rifaximin (AR), before placebo (BP), and after placebo (AP).
Figure 2
Figure 2
Modulation of TLR2 and TLR4 lymphocytes in PBMC and gut. (a, b) The comparison of modulation of TLR2 and TLR4 expression in the different patient groups between PBMC and gut (before and after placebo or Rifaximin). (c-d) Modulation of TLR2 and TLR4 expression observed in a single representative patient. The values reported in box-and-whiskers plots show the minimum, the 25th percentile, the median, the 75th percentile, and the maximum. In particular, the whiskers go down to the smallest value (minimum) and up to the largest one (maximum), the top and bottom of the box are the 25th and 75th percentiles, and the line in the middle of the box is the median, corresponding to 50th percentile, evaluated before Rifaximin (BR), after Rifaximin (AR), before placebo (BP), and after placebo (AP).
Figure 3
Figure 3
Expression of TLRs in monocytes and granulocytes. (a, b) Percentage of TLR2 and TLR4 monocytes in peripheral blood, at baseline and after Rifaximin and placebo treatments. (c, d) Median of fluorescence (MFI) of TLR2 and TLR4 monocytes in peripheral blood at baseline and after Rifaximin and placebo treatments. (e, f) MFI of TLR2 and TLR4 granulocytes in peripheral blood at baseline and after Rifaximin and placebo treatments. The values reported in box-and-whiskers plots show the minimum, the 25th percentile, the median, the 75th percentile, and the maximum. In particular, the whiskers go down to the smallest value (minimum) and up to the largest one (maximum), the top and bottom of the box are the 25th and 75th percentiles, and the line in the middle of the box is the median corresponding to 50th percentile (***P < 0.001, **P < 0.01, and *P < 0.05 patients versus controls (health or baseline)). Healthy control (CTR), UDD patients (PZ), before Rifaximin (BR), after Rifaximin (AR), before placebo (BP), and after placebo (AP).
Figure 4
Figure 4
Expression of CD103 in lymphocytes of sigma mucosa. (a) Percentage of CD103 in CD4 lymphocytes subpopulation after Rifaximin and placebo treatments. (b) Percentage of CD103 in CD8 lymphocytes subpopulation after Rifaximin and placebo treatments. (c) Percentage of TCR-gamma/delta CD103 lymphocytes subpopulation after Rifaximin and placebo treatments. The values reported in box-and-whiskers plots show the minimum, the 25th percentile, the median, the 75th percentile, and the maximum. In particular, the whiskers go down to the smallest value (minimum) and up to the largest one (maximum), the top and bottom of the box are the 25th and 75th percentiles, and the line in the middle of the box is the median corresponding to 50th percentile (*P < 0.05 before versus after treatments in patients); before Rifaximin (BR), after Rifaximin (AR), before placebo (BP), and after placebo (AP).

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