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
Abstract
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.

AIM

To elucidate whether gut microbiota and immune system changes appear following anti TNFα therapy during dextran sulfate sodium (DSS) colitis.

METHODS

Eighty C57BL/6 mice were divided into four groups: “No DSS”, “No DSS + anti-TNFα”, “DSS” and “DSS + anti-TNFα”. “DSS” and “DSS + anti-TNFα” were treated for 5 d with 3% DSS. At day 3, mice whithin “No DSS+anti-TNFα” and “DSS+anti-TNFα” group received 5 mg/kg of an anti-TNFα agent. Forty mice were sacrificed at day 5, forty at day 12, after one week of recovery post DSS. The severity of colitis was assessed by a clinical score (Disease Activity Index), colon length and histology. Bacteria such as Bacteroides, Clostridiaceae, Enterococcaceae and Fecalibacterium prausnitzii (F. prausnitzii) were evaluated by quantitative PCR. Type 1 helper T lymphocytes (Th1), type 17 helper T lymphocytes (Th17) and CD4+ regulatory T lymphocytes (Treg) distributions in the mesenteric lymph node (MLN) were studied by flow cytometry.

RESULTS

Bacteria associated with a healthy state (i.e., such as Bacteroides, Clostridiaceae and F. prausnitzii) decreased during colitis and increased in course of anti-TNFα treatment. Conversely, microorganisms belonging to Enterococcaceae genera, which are linked to inflammatory processes, showed an opposite trend. Furthermore, in colitic mice treated with anti-TNFα microbial changes were associated with an initial increase (day 5 of the colitis) in Treg cells and a consequent decrease (day 12 post DSS) in Th1 and Th17 frequency cells. Healthy mice treated with anti-TNFα showed the same histological, microbial and immune features of untreated colitic mice. “No DSS + anti-TNFα” group showed a lymphomononuclear infiltrate both at 5th and 12th d at hematoxylin and eosin staining, an increase of in Th1 and Th17 frequency at day 12, an increase of Enterococcaceae at day 5, a decrease of Bacteroides and Clostridiaceae at day 12.

CONCLUSION

Anti-TNFα treatment in experimental model of colitis improves disease activity but it is associated to an increase in Th17 pathway together with gut microbiota alteration.

Keywords: Gut microbiota; Dextran sodium sulphate colitis; Immune system; T cells; Mesenchymal lymphnode; Tumor necrosis factor α

Core tip: The effect of the gut microbiota on the immune responses is not limited to the gut. On the other hand, autoimmune diseases, such as inflammatory bowel disease, are associated to different degree of dysbiosis involving different bacterial genera. Using a colitic mouse model, we aimed to evaluate the impact of anti-tumor necrosis factor α (TNFα) therapy on the intestinal immune system and gut microbiota concurrently. Healthy mice treated with anti-TNFα showed similar histological, microbial and immune features of untreated colitic mice, a lymphomononuclear infiltrate both at day 5 and 12 at hematoxylin and eosin staining, an increase of type 1 helper T lymphocytes and type 17 helper T lymphocytes at day 12, and finally increase of Enterococcaceae at day 5, a decrease of Bacteroides and Clostridiaceae at day 12.