Basic Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Mar 28, 2019; 25(12): 1465-1477
Published online Mar 28, 2019. doi: 10.3748/wjg.v25.i12.1465
Anti-tumor necrosis factor α therapy associates to type 17 helper T lymphocytes immunological shift and significant microbial changes in dextran sodium sulphate colitis
Valentina Petito, Cristina Graziani, Loris R Lopetuso, Marco Fossati, Alessandra Battaglia, Vincenzo Arena, Domenico Scannone, Gianluca Quaranta, Andrea Quagliariello, Federica Del Chierico, Lorenza Putignani, Luca Masucci, Maurizio Sanguinetti, Alessandro Sgambato, Antonio Gasbarrini, Franco Scaldaferri
Valentina Petito, Loris R Lopetuso, Antonio Gasbarrini, Franco Scaldaferri, Istituto di Patologia Speciale Medica, Università Cattolica del Sacro Cuore, Roma 00168, Italy
Cristina Graziani, Loris R Lopetuso, Antonio Gasbarrini, Franco Scaldaferri, UOC di Medicina Interna e Gastroenterologia, Area di Gastroenterologia e Oncologia Medica, Dipartimento di Scienze Gastroenterologiche, Endocrino-Metaboliche e Nefro-Urologiche, Fondazione Policlinico Universitario “A. Gemelli” IRCCS, Roma 00168, Italy
Marco Fossati, Dipartimento delle Scienze della Salute della Donna e del Bambino, Fondazione Policlinico Universitario “A. Gemelli” IRCCS, Roma 00168, Italy
Alessandra Battaglia, Istituto di Clinica Ostetrica e Ginecologica, Università Cattolica del Sacro Cuore, Roma 00168, Italy
Vincenzo Arena, U.O.S.A. Gineco-Patologia e Patologia Mammaria, Fondazione Policlinico Universitario “A. Gemelli” IRCCS, Roma 00168, Italy
Vincenzo Arena, Istituto di Anatomia Patologica, Università Cattolica del Sacro Cuore, Roma 00168, Italy
Domenico Scannone, Dipartimento di Anatomia Patologica e Istologia, Fondazione Policlinico Universitario “A. Gemelli” IRCCS, Roma 00168, Italy
Gianluca Quaranta, Luca Masucci, Maurizio Sanguinetti, Dipartimento di Microbiologia, Fondazione Policlinico Universitario “A. Gemelli” IRCCS, Roma 00168, Italy
Gianluca Quaranta, Luca Masucci, Maurizio Sanguinetti, Istituto di Microbiologia, Università Cattolica del Sacro Cuore, Roma 00168, Italy
Andrea Quagliariello, Federica Del Chierico, Lorenza Putignani, Unità di Microbioma Umano, Ospedale Pediatrico Bambino Gesù IRCCS, Roma 00146, Italy
Lorenza Putignani, Unità di Parassitologia, Ospedale Pediatrico Bambino Gesù IRCCS, Roma 00146, Italy
Alessandro Sgambato, Istituto di Patologia Generale, Università Cattolica del Sacro Cuore, Roma 00168, Italy
Author contributions: Petito V and Scaldaferri F designed the research. Petito V performed the majority of the experiments in vivo and in vitro, and analyzed the data; Graziani C and Lopetuso LR gave their support in experiments with animals. Fossati M and Battaglia A helped to design cytometry experiments. Arena V scored the histology slides. Scannone D prepared histological slides and stained the slides of mice colon. Quaranta G supported the qPCR analysis. Quagliariello A and Del Chierico F performed the statistical and biostatistical analysis. Putignani L, Masucci L, Sanguinetti M and Sgambato A made critical revisions related to important intellectual content of the manuscript. Gasbarrini A and Scaldaferri F gave the final approval of the article to be published.
Institutional animal care and use committee statement: This manuscript was approved by the institutional animal care and use committee, No. NN42 (25/11/2013-25/11/2016).
Conflict-of-interest statement: Authors disclose no any conflict of interest.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: it was uploaded on Novembre 11th, 2018
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Loris R Lopetuso, MD, PhD, Academic Fellow, Senior Postdoctoral Fellow, UOC di Medicina Interna e Gastroenterologia, Area di Gastroenterologia e Oncologia Medica, Dipartimento di Scienze Gastroenterologiche, Endocrino-Metaboliche e Nefro-Urologiche, Fondazione Policlinico Universitario “A. Gemelli” IRCCS, Largo A Gemelli 8, Roma 00168, Italy. lopetusoloris@libero.it
Telephone: +39-329-3137604
Received: November 14, 2018
Peer-review started: November 14, 2018
First decision: January 6, 2019
Revised: February 15, 2019
Accepted: February 22, 2019
Article in press: February 23, 2019
Published online: March 28, 2019
Processing time: 134 Days and 19.8 Hours
ARTICLE HIGHLIGHTS
Research background

Anti-tumor necrosis factor α (TNFα) represents the best therapeutic option to induce mucosal healing and clinical remission in patients with moderate-severe ulcerative colitis. On the other side gut microbiota plays a crucial role in pathogenesis of ulcerative colitis but few information exists on how microbiota changes following anti-TNFα therapy and on microbiota role in mucosal healing.

Research motivation

The hypothesis behind this study is that anti-TNFα could induce a dysbiosis sustained by immunological changes: dysbiosis could represent one of the reasons for loss of response to anti-TNFα therapy or primary failure.

Research objectives

With this manuscript we aimed to evaluate, in colitic mice as well as healthy mice, intestinal immune system status and gut microbiota modulation induced by anti-TNFα therapy. A more comprehensive approach including gut microbiota modulation, if clarified, could be assessed by dedicated studies on active gut microbiota modulation during biologic therapy in inflammatory bowel disease (IBD).

Research methods

Healthy mice treated with anti-TNFα showed similar histological, microbial and immune features of untreated colitic mice, in particular a lymphomononuclear infiltrate both at 5th and 12th d at hematoxylin and eosin staining, an increase of type 1 helper T lymphocytes (Th1) and type 17 helper T lymphocytes (Th17) at day 12, and finally increase of Enterococcaceae at day 5, a decrease of Bacteroides and Clostridiaceae at day 12.

These findings are particularly relevant to understand the role that anti-TNFα modulation plays on gut microbiota (and vice-versa) also in humans: this finding could be of major interest in order to give more lights on mechanisms of loss of response to anti-TNFα, mechanisms of immunological shift with towards other pathways, like Th17 pathways, as well as novel potential therapeutic targets in IBD.

Research results

Gut microbiota contributes to immune system priming, development and activation. In course of IBD a decrease of Firmicutes (and, among these, the species Faecalibacterium prausnitzii) together with an increase in Proteobacteria has been demonstrated, associated to an increase of T lymphocyte differentiation toward Th1, Th17 subsets and a decrease of CD4+ regulatory T lymphocytes (Tregs). How these conditions are modified in course of medical treatment is not well clarified, particularly when a powerful modulator of the immune system, like anti-TNFα, is utilized.

Research conclusions

Our study confirmed data from the literature. The same features were confirmed in the experimental model of colitis presented in this paper. Furthermore, it was shown that higher representation of Bacteroides, Clostridiaceae and Faecalibacterium prausnitzii and lower representation of Enterococcaceae are associated to a healthy state while the opposite happens during colitis. Anti-TNFα is able to induce changing in gut microbiota toward a health state in course of active colitis, but a more dysbiotic effect when utilized in healthy controls. Healthy related-microbial assessment is associated to higher Treg cells count and lower Th1 and Th17 frequency, condition modified in course of active colitis or use of anti-TNFα in course of healthy condition.

The study conclusion demonstrates a clear relationship between microbial changes and immunological changes in healthy as well as in controls. However, whether these findings correlate to human disease needs to be assessed in a dedicated study as well the need of checking and correction of dysbiosis in patients exposed to anti-TNFα.

Research perspectives

This study strongly suggests a complete assessment of immune system and microbiota in course of powerful immunomodulatory therapy like anti-TNFα and opens new and important considerations. In order to personalize the therapeutic approach to an IBD patient, clinical studies assessing gut microbiota composition before starting an immunomodulatory therapy and in course of therapy are needed. Once an alteration has been demonstrated, the potential role of concomitant microbiota modulation could also be considered for newer and more personalized therapeutic approaches.