Accepted Manuscript

Title: Actinobacteria: a relevant minority for the maintenance

of gut homeostasis

Author: Cecilia Binda Loris Riccardo Lopetuso Gianenrico

Rizzatti Giulia Gibiino Vincenzo Cennamo Antonio
Gasbarrini

PII: S1590-8658(18)30210-X
DOI: https://doi.org/doi:10.1016/j.dld.2018.02.012
Reference: YDLD 3676

To appear in: Digestive and Liver Disease

Received date: 18-10-2017

Revised date: 26-1-2018
Accepted date: 19-2-2018

Please cite this article as: Binda C, Lopetuso LR, Rizzatti G, Gibiino
G, Cennamo V, Gasbarrini A, Actinobacteria: a relevant minority for the
maintenance of gut homeostasis, Digestive and Liver Disease (2018),
https://doi.org/10.1016/j.dld.2018.02.012

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Title page

ACTINOBACTERIA: A RELEVANT MINORITY FOR THE MAINTENANCE OF GUT

HOMEOSTASIS

Cecilia Binda*, Loris Riccardo Lopetuso*, Gianenrico Rizzatti*, Giulia Gibiino*, Vincenzo

Cennamo^, Antonio Gasbarrini*

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* Department of Internal Medicine, Gastroenterology and Hepatology, Catholic University of

Sacred Heart of Rome, A. Gemelli Hospital - Italy.

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^ Unit of Gastroenterology and Digestive Endoscopy, AUSL Bologna Bellaria-Maggiore Hospital,

Bologna - Italy
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Electronic word count: 4120
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Corresponding Author:
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Prof. Antonio Gasbarrini

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Department of Internal Medicine, Gastroenterology Division,

Catholic University of Rome, Policlinico “A. Gemelli” Hospital

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Largo Gemelli, 8
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00168 Rome (ITALY)

Phone/fax number: 0039-06-30156018

E-mail: antonio.gasbarrini@unicatt.it

Page 1 of 36
Abstract

Actinobacteria are one the four major phyla of the gut microbiota and, although they represent only

a small percentage, are pivotal in the maintenance of gut homeostasis. During the last decade many

studies focused the attention on Actinobacteria, especially on their role both in gastrointestinal and

systemic diseases and on their possible therapeutic use. In fact, classes of this phylum, especially

Bifidobacteria, are widely used as probiotic demonstrating beneficial effects in many pathological

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conditions, even if larger in vivo studies are needed to confirm such encouraging results. This

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review aims to explore the current knowledge on their physiological functions and to speculate on

their possible therapeutic role(s) in gastrointestinal and systemic diseases.

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Key words: gut microbiota, Actinobacteria, Bifidobacteria spp, gut homeostasis, dysbiosis,

probiotic.
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INTRODUCTION

The gut microbiota is composed by multiple commensal microbial species including 100 trillion

(10^14) bacteria, quadrillion viruses, fungi, parasites, archeas and yeasts, reaching an overall

biomass of about 1 kg and more than 3 million of genes [1- 4]. The different gastrointestinal regions

are characterized by a different bio-compartmentalization with a distinct and stable microbial

community. This is influenced by the acid environment, the presence of bile and pancreatic

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secretion and the high peristaltic activity in the stomach and small intestine, which do not allow a

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stable bacterial colonization, differently from the colon where bacterial colonization is favored by

the low redox potential and the slow transit [5, 6]. The majority of microbes forming the human

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microbiota can be assigned to four major phyla: Bacteroidetes, Firmicutes, Proteobacteria and

Actinobacteria [7, 8]. Firmicutes and Bacteroidetes represent more than 90% of the relative
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abundance of the gut microbiome and their relationship plays a pivotal role in the maintenance of
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gut homeostasis. Actinobacteria and Proteobacteria represent the remaining 10% [7, 8].

Mainly oropharyngeal origin aerobic gram-positive bacteria inhabit stomach, duodenum and
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jejunum, whereas coliforms and anaerobic species (such as Bacteroides, Bifidobateria, Clostridia
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and Lactobacilli) are predominant in the ileum and post ileocecal valve, respectively [9, 10].
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Gut microbiota is involved in many useful functions, such as energy production from nutrient

biotransformation [11, 12], regulation of lipid metabolism [13], metabolism of vitamins and
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absorption of calcium, magnesium and iron [12, 14, 15], maintenance of the intestinal barrier
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function [16, 17], the development of immune system from the first days of life [18- 23].

Many conditions are supposed to be related to quantitative and qualitative changes in gut

microbiota composition and function, such as inflammatory bowel diseases, celiac disease, irritable

bowel syndrome (IBS), obesity, non-alcoholic fatty liver disease (NAFLD) and non-alcoholic

steato-hepatitis (NASH), diabetes, cardiovascular disease, arthritis, psoriasis and psychiatric

disorders. This gut microbiota impairment is known as dysbiosis [24- 30].

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Recently, increasing interest has focused on the phyla of Actinobacteria, especially on

Bifidobacteria family. The purpose of this review is to explore the crucial role of Actinobacteria in

the maintenance of gut homeostasis and the critical importance of their future therapeutic

applications.

ACTINOBACTERIA IN THE GASTROINTESTINAL TRACT

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Actinobacteria are Gram positive, multiple branching rods, non-motile, non-spore-forming and

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anaerobic bacteria [31], that include three main anaerobe families (Bifidobacteria, Propionibacteria

and Corynebacteria) and an aerobe family (Streptomyces). The most represented in the human gut

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are Bifidobacteria. Interestingly, the complete genome sequences are available for certain

Bifidobacterial species, like B. adolescents, B. animalis, B. breve, B. bifidum, B. long and B.

angulatum [31].
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Many factors can impact on the presence of Actinobacteria in the intestine. Although the relation

between phylum diversity and delivery is still unclear, studies demonstrated a higher diversity
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within Actinobacteria phylum among children that underwent vaginal delivery [32-37]. In fact,
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phylum diversity within Actinobacteria, (i.e., Bifidobacterium) was significantly lower in infants
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delivered by caesarean section compared to vaginally delivered newborns during the first week of

life [33 -35], at the age of 1 month [36], and three months [37]. Other studies did not confirm such
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differences or showed a lower effect on Bifidobacteria and Bacteroides colonization at the age of 6
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and 12 month [38, 39]. The lower abundance of Bifidobacteria and Bacteroides in c-section

delivered infants may be explained with a higher mother consumption of antibiotics before, during

and after the delivery. Indeed, postnatal use of antibiotic has been associated with decreased

numbers of Bifidobacterium and Bacteroides and a higher relative abundance of the Clostridium

leptum [40- 42]. Furthermore, a higher abundance of Bifidobacterium genus has been demonstrated

in breastfed infants [43]. Interestingly, it has been demonstrated that human milk is rich in

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substances, like human milk oligosaccharides (HMOs), that may act as probiotics and stimulate the

growth of Lactobacillus and Bifidobacterium species [44].

Weaning causes a further change in microbial composition and Bifidobacteria are no longer the

dominant group, with the adult microbiota established after the second year of life [45]. During the

adulthood, the percentage of Actinobacteria remains stable around 8% [46] and constitutes one of

the prevalent commensal bacterial in the distal small and large intestine [47]. Intriguingly, an Italian

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study on centenarian patients showed a reduction in the total number of anaerobes compared to

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young adults, with a reduction of Bifidobacteria and Bacteroides [48]. These evidences have opened

new horizons for the concept of “fragile” microbiota that could deeply affect the delicate health

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equilibrium in old patients.

ACTINOBACTERIA AND GUT BARRIER

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Gut barrier is a multi-layer system able to afford a daily exposure to pathogens and external

environment. It can be divided into a superficial physical barrier constituted by the epithelial cells,
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the tight junctions and the mucus; and a functional barrier represented by gut-associated mucosa
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lymphoid tissue (GALT), peristalsis and antimicrobial substances, able to regulate the
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immunological response to pathogens and tolerance to commensal bacteria [16, 49]. Gut microbiota

is pivotal in the maintenance of intestinal barrier functions, increasing tight junctions expression,
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regulating mucin biosynthesis and catabolism, providing energy for epithelial cells proliferation and
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stimulating the immune system [17, 49]. In this scenario, Actinobacteria are absolute players in

maintaining gut barrier homeostasis.

The production of short chain fatty acids (SCFA), such as acetate, propionate and butyrate, from

carbohydrate fermentation is crucial for providing energy to epithelial cells turnover and for their

potent antibacterial activity [16, 17]. In this field, Bifidobacteria have beneficial effects in the

maintenance of gut barrier thanks to their great production of SCFA [50]. In particular, they

produce high concentration of acetate that can protect the host from enteropathogenic infections,

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such as entero-hemorrhagic Escherichia coli and Shigella [51]. Moreover it has been demonstrated

that Bifidobacteria have a non-negligible production of lactate, which can be metabolized by a

group of bacteria (“lactate utilizer”) to produce butyrate [52]. In this scenario, in vitro studies

demonstrated that SCFA, especially butyrate, are correlated with an increased expression of the

gene MUC2, a mucin glycoprotein that is one of the major component of the mucous layer and thus

of intestinal barrier [53-55]. The mechanism responsible for the increased production of MUC

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induced by butyrate is not definitely established. Gaudier et al demonstrated that butyrate could

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modify MUC gene expression in goblet cells depending on the energy source available with a

consequent higher expression of MUC2 in conditions of glucose deprivation when the consumption

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of butyrate by goblet cells is increased [54]. Authors concluded that butyrate metabolism is

involved not only in mucin synthesis but can play a role also in gene transcription. Another in vitro
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study showed a relationship between SCFA, especially butyrate, the production of prostaglandins
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(PG) by intestinal myofibroblasts and epithelial mucin expression [53]. In fact, SCFA seemed to

enhance the production of PGE1 in subepithelial myofibroblasts that in turn was able to stimulate
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epithelial mucin expression, probably through the epithelial differentiation induced by cell-to-cell
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contact, extracellular matrix and soluble factors such as transforming growth factor β. Other
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important elements that contribute to the viscoelastic properties of the mucous layer are the Trefoil

factors (TFFs). These mucin-associated peptides are mainly secreted by goblet cells, are supposed
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to be involved in the mucosal maintenance and repair, and seem to reduce the recruitment of
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inflammatory cells [56, 57]. Moreover, butyrate contributes to the colonic defense barrier by

increasing the expression of TFFs [58]. In vitro and in vivo studies and in studies showed a possible

effects of butyrate on the expression of heat shot proteins (HSPs), which contribute to the gut

barrier homeostasis by suppressing the production of inflammatory modulators [59, 60]. A

mechanism for the enhancement of HSPs expression has not been already discerned. So far, only

some hypotheses have been proposed. Indeed, SCFA are able to induce cellular acidification

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through a non-ionic diffusion that may induce the production of stress kinases and HSPs. Moreover,

the butyrate seems to inhibit the histone deacetylase, a crucial transcriptional regulator [59].

Finally, a possible role of butyrate on intestinal permeability has been assessed in different in vitro

studies, which demonstrated a butyrate influence on tight junctions expression. Interestingly, these

effects seem to be concentration-dependent, with a concentration up to 2 mM that can decrease the

intestinal permeability and a concentration higher than 8 mM that, on the contrary, is able to induce

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an increased permeability [56, 61, 62].

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ACTINOBACTERIA AND METABOLISM

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Human intestine does not possess many of the enzymes involved in the degradation and

biotransformation of substances introduced with the diet. Indeed, the majority of enzymes engaged
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in energy production is provided by the commensal bacteria, especially located in the colon and
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belonging to the genus Bacteroides, Bifidobacterium, Ruminococcus and Roseburia [63]. In

particular, the pathways involved include the fermentation of large polysaccharides,

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oligosaccarides, unabsorbed sugars and fibers, that release hydrogen, carbon dioxide and SCFAs,
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the degradation of proteins, the regulation of lipid metabolism through lipoprotein lipase (LPL), and
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the absorption and biosynthesis of vitamin K, B12, iron, calcium and magnesium [13-15, 64-67].

Bifidobacteria are able to produce large quantities of acetate, but do not produce propionate and
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butyrate, which are mainly produced by Bacteroides phylium and Clostridium cluster XIVa and IV
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[68, 69]. However, the acetate released by B. longum NCC2705 represents a co-substrate for

butyrate production [70] that constitutes the main energy source for colonocytes [71].

Actinobacteria are also involved in the biodegradation of resistant starch [72 - 74]. Actinobacteria,

in particular Bifidobacteria, through the glycosyl hydrolases (GHs) hydrolyze the glycosidic bond

between two or more sugars and cooperate to the breakdown of plant-derived carbohydrate starch

and polysaccharides, such as FOS, GOS, XOS, inulin or arabinoxilan [75 - 77]. Moreover,

Bifidobacteria are supposed to be involved in the transformation of linoleic acid (LA) into

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conjugated linoleic acids (CLA), a group of isomers of LA [78], that have potential health-

promoting properties like anticarcinogenesis, anti-atherosclerosis, anti-diabetes, anti obesity and

enhancement of immune functions [79].

Although it has been largely demonstrated that the diet strongly influences gut microbiota

composition, the relationship between diet and Actinobacteria is still unclear. In fact, energy

restrictions, high fiber diet and dietary components such as fructo-oligosaccharides (FOS), galacto-

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oligosaccharides (GOS), xylo-oligosaccharides (XOS) are associated to higher microbial diversity

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[80]. While some studies showed that Actinobacteria abundance is positively associated with a

high-fat diet and negatively associated with fiber intake [81, 82], an opposite association is

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suggested by other studies in which a high concentration of Bifidobacterium spp is positively

correlated with lean individuals, high consumption of complex carbohydrates, improvement of

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glucose homeostasis, reduction of obesity and inflammation [83 – 86]. A study comparing fecal
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samples of lean and obese women showed different concentration of Bifidobacteria and Clostridium

coccoides between the two groups. In particular, there was a negative correlation between
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Bifidobacteria and body fat percentage with an inverse correlation with insulin levels and the
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homeostasis model assessment (HOMA) index [83]. However, in this study there was no difference
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in LPS levels between lean and obese subjects and no correlation with body fat percentage, insulin

levels or HOMA-index. Nevertheless, a study by Cani et al [85] demonstrated that mice fed with an
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high fat diet enriched with prebiotic (fermentable dietary fibre, oligofructose (OFS)) could restore
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their levels of Bifidobacteria, improve glucose tolerance, insulin secretion, and induce an increase

of proglucagon mRNA precursor. Authors highlighted that all these changes were correlated with a

reduction of endotoxiemia, suggesting a protective role of Bifidobacteria, while no relationship was

evident with other bacterial group [85]. Despite the mechanism of action has not been completely

understood, authors suggested that the product of OFS degradation, the SCFA, could ameliorate the

gut barrier function both through a direct action on colonic cells and by modulating gut microbiota,

especially Bifidobacteria [85]. Furthermore, the use of dextrins derived from maize starch is able to

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stimulate the growth of Actinobacteria and Bacteroidetes in both normal-weight and obese children

[87], with possible beneficial effect on the production of SCFA.

A modulation on gut microbiota, and therefore on Actinobacteria phylum, has been shown with the

diet low in fermentable oligosaccharides, disaccharides and polyols, fructo-oligosaccharides and

galacto-oligosaccharides (FODMAPs). FODMAPs are small carbohydrate molecules that are

slowly absorbed in the form of monosaccharides or not absorbed when disaccharides or

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polysaccharides, lasting for a prolonged period into the intestinal lumen [67]. There, FODMAPs

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exert an osmotic action collecting water into the lumen and are fermented by commensal bacteria

releasing SCFA, dioxide, hydrogen, methane and carbon [88, 89]. A study by McIntosh et al

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compared the impact of high and low FODMAPs diet in IBS patients and demonstrated an

increased richness of Actinobacteria with a decreased number of Bifidobacteria in the low

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FODMAPs diet group [90]. On the contrary, other studies showed that a low FODMAPs diet could
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lead to a lower abundance in both Bifidobacteria and Actinobacteria because of their FODMAPs

need for their metabolism [91, 92]. However, not all the patients treated with low FODMAPs
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experienced an improvement of IBS symptoms [90]. This could probably due to different effects of
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diverse Bifidobacteria strains. Recent studies demonstrated that the use of Bifidobacterium infantis
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and Bifidobacterium animalis has beneficial effects on IBS symptoms [93 - 95]. However, all the

available studies have not examined the microbiome of patients with IBS following the
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reintroduction phase, but only after the end of the strict low-FODMAP diet, which is usually only
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recommended for 2 to 6 weeks.

ACTINOBACTERIA AND IMMUNOLOGICAL FUNCTION

The gut microbiota plays a pivotal role in the development of immune system and Bifidobacteria,

are also critical in this field. The stimulation of intraepithelial lymphocytes, the production of

mucosal immunoglobulins and the promotion of a tolerogenic immune response are the main

functions exerted by commensal bacteria [19, 20,22, 23, 96].

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A decreased number of Bifidobacteria is associated to an enhancement of gut permeability [97] that

leads to the translocation of LPS into the serum. This triggers the immune system activation and

sustains chronic inflammatory conditions, such as insulin resistance, diabetes and liver diseases

[98]. The administration of Bifidobacterium pseudocatenulatum CECT 7765 along with high fat

diet in mice is able to down-regulate the inflammation by reducing the production of inflammatory

cytokines and chemokines, especially IL-6 and MCP-1, which are usually increased in obesity and

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metabolic disorders [99]. The reduction of these inflammatory markers is concomitant with the

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improvement of glucose tolerance and with the increasing of IL-4 and IL-13. These cytokines are

able to stimulate the M2 phenotypic differentiation of macrophages, a subtype of adipose tissue

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macrophages that secretes the anti-inflammatory cytokine IL-10, and to promote the control of

inflammation and normal insulin sensitivity [100]. However, the same study showed that after the
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administration of B. pseudocatenulatum CECT 7765 there were increased level of IL-10 only in
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standard diet fed mice and not in high fat diet-fed mice, indicating that probably parallel regulatory

mechanisms are involved. Furthermore, B. pseudocatenulatum CECT 7765 can boost an

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appropriate inflammatory response to a bacterial stimulus (LPS) that is usually impaired in high fat
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diet fed mice. In fact, the administration of this probiotic in these mice stimulates the production of
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TNF-α by LPS-stimulated macrophages, boosts the oxidative burst and therefore the phagocytosis

function in peritoneal macrophages. Finally, it increases the ability of DCs to present antigens and
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prime a T-lymphocyte proliferative response [99]. In this direction, the reduced phagocytic capacity
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and oxidative burst of macrophages and the impaired function of DCs are possible reasons for the

increased susceptibility to infections in obese subjects [101, 102].

Moreover, Actinobacteria, and mainly Bifidobacteria species, can modulate immune-inflammatory

and autoimmune response by inducing regulatory T-cells [103, 104].

Grounding on the evidence that Bifidobacterium longum subspecies infantis (B.infantis) induces

regulatory T cells activity in animal models [103, 105 - 108] and increases the relative proportion of

the same cells in peripheral blood of healthy humans [109], Groeger et al studied the effect of

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B.infantis inflammatory mediator production in inflammatory disorders in patients with ulcerative

colitis (UC), psoriasis and chronic fatigue syndrome (CFS) [110]. They found that 6-8 weeks of

treatment with B.infantis significantly reduced plasma C-reactive protein (CRP) levels in all three

group of patients, whereas a statistically significant attenuation of TNF-α was observed for psoriasis

and CFS. In the UC group they observed a trend of reduction in TNF-α levels that however did not

reach the statistical significance, probably because of the shorter treatment period (6 weeks of

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treatment vs 8 weeks of treatment of the other two groups). Instead, plasma levels of IL-6 were only

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marginally decreased after treatment reinforcing the hypothesis of a specific influence on immune

response for each gut microbiota component [110]. The use of Bifidobacterium species has also

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been studied in respiratory pathology, alone or in combination with prebiotic or other probiotics like

Lactobacilllus. In this field, Bifidobacteria seem to be effective to prevent asthma-like symptoms in

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infant with atopic dermatitis and to treat the symptoms of allergic rhinitis [111 – 115]. This is
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probably due to an immunomodulatory effect of Bifidobacteria on Th balance, regulating Th-2

immunoresponse that is usually skewed in atopic patients and is responsible of higher levels of
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eosinophils and IgE production [113].

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ACTINOBACTERIA AND GUT-BRAIN AXIS

Recently, the interest on the impact of gut microflora on gut-brain axis is increasing. This could
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involve neuroendocrine, immunological and direct neural mechanisms [116 - 118]. Dysbiosis and
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the consequent increased intestinal permeability are associated to an up-regulation of systemic

inflammation that may also involve the central nervous system [119]. Both the gut microbiota

production of neurochemicals (i.e., serotonin, SCFA, dopamine and γ-aminobutyric acid) [116, 120]

and neurotoxic metabolites (i.e., ammonia) [119] by strengthens the possibility of an implication of

the gut-brain axis in depression, anxiety, IBS, IBD, and neurodevelopmental disorders such as

autism [116, 121 - 124]. A direct neural communication between the gut and the brain through the

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stimulation of the enteric nervous system and consequently the vagus nerve has also been supposed

[125, 126].

Standing on previous studies that showed a reduction of Bifidobacteria and lactobacilli following a

stressful experience and emotional stress both in animals and humans [127 - 131], Desbonnet et al

conducted a study to assess the potential antidepressant properties of probiotic Bifidobacteria

infantis in rats [132]. They demonstrated that a two-week treatment with the probiotic

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Bifidobacteria infantis determines an attenuation of pro-inflammatory immune response and an

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elevation of tryptophan, a precursor of serotonine that represents a target of antidepressant

treatments. Authors concluded that these results might support the beneficial effect of this probiotic

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in the treatment of depression, especially when associated to gastrointestinal disorders. Two years

later, these findings were supported by the evidence that the chronic treatment with Bifidobacteria
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infantis was able to attenuate depression in the rats exposed to maternal separation stress in early
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life [133].

On the other side, a recent study by Lyte at al [134] showed that mice fed with resistant starch (RS)
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had increased levels of Bifidobacteria, according also to a previous study by Tachon et al [135].
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RS-fed mice evidenced a significant increase in anxiety-like behavior that the authors considered to
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be induced by the same diet with a concomitant abundance of Bifidobacterium [134].

Furthermore, a role of Actinobacteria, mainly Bifidobacteria, in the deterioration of cognitive

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function has been supposed. In fact, preliminary data on the use of the probiotic VSL#3, a mixture
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of 8 different strains of bacteria including three Bifidobacteria species, showed an improvement in

the age-related decrease of long-term potentiation (LTP) in rats [136]. One of the mechanisms

supposed to be involved in the regulation of LTP is the anti-inflammatory effect of VSL#3 on the

modulation of the hippocampal molecules expression, such as CD 68 and CD 11b. These are

markers of microglial activation, and therefore of inflammation. Moreover the authors reported that

VSL#3 is able to influence the expression of several genes in the cortex. In particular, it is able to

attenuate the age-related changes in three genes, PLA2G3, Nid2 and Alox15, which are associated

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to inflammation. Changes of genes expression in the brain may be linked to the variation of several

molecules induced by VSL#3. In fact, this study showed that this probiotic regulates specific

mediators involved in synaptic plasticity in the hippocampus of aged rats, especially brain-derived

neurotrophic factor (BDNF), synapsin and syntaxin. The authors suggested that the synaptotrophic

effect of VSL#3 is mainly due to the BDNF, whose expression was increased in those rats treated

with the probiotic. The increase of BDNF levels was associated to higher level of synapsin. Of note,

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previous studies recognized its fundamental role in sustaining LTP through the induction of

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neurogenesis and synaptogenesis [136].

Lastly, the existence and the importance of gut-brain axis have also been suggested in alcohol

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dependence. Some alcohol dependent subjects develop a high intestinal permeability that is

associated with significantly lower levels of Bifidobacteria [137]. These subjects are characterized
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by higher levels of depression, anxiety and craving and have a consistent higher risk of persistence
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of psychological symptoms after detoxification when compared to alcohol dependent subject with a

lower intestinal permeability, that on the contrary recover completely after the abstinence period.
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BIFIDOBACTERIA AS PROBIOTIC
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Bifidobacteria, together with Lactobacilli, represents the cornerstone of probiotic therapeutic

approach, but only few Bifidobacteria strains have been studied alone. Thus, in the latter cases the
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beneficial effects cannot be exclusively related to this probiotic family.

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For example, the probiotic formulation VSL#3, a mixture of lyophilized four Lactobacilli (L. casei,

L. plantarium, L acidophilus, L. bulgaricus), three Bifidobacteria strains (B. longuum, B. breve, B.

infantis) and Streptococcus salivaris subspecies termophilus demonstrated to be effective in several

conditions. Its efficacy has been mainly assessed in the prevention of pouchitis relapse in patients

with UC [138, 139], with promising results in maintaining remission and treating mild-to-moderate

UC both in adult and children [140 - 143], in metabolic disorders, in non-alcoholic steato-hepatitis

(NASH) [144, 145] and in the prevention of antibiotic-associated diarrhea [146]. An amelioration of

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the pro-inflammatory status in patients affected by UC has been demonstrated with the use of

Bifidobacterium infantis alone [110], as mentioned above, and Bifidobacterium breve in association

with GOS [147]. The use of Bifidobacteria, alone or in association with other probiotics is also

effective in the alleviation of symptoms in patients with IBS [93 - 95, 148 - 150], constipation [151,

152] and in the reduction of antibiotic-associated diarrhea in infants [153]. Moreover, a recent study

showed that a supplementation of B.breve in association with breast feeding is able to reduce the

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incidence of necrotizing enterocolitis (NEC) in preterm neonates born between 28 and 34 weeks of

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gestation. However, further studies are needed for neonates < 28 weeks where the reduction of NEC

didn’t reach a statistical significance [154].

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Furthermore, in vitro and murine models studies highlighted how Bifidobacteria, especially B.

longum and B. lactis strains, seem to be active against rotavirus infection [155] and could protect
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against colorectal cancer (CRC) [156, 157]. Grounding on a previous study where the use of a
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combination of RS and B. lactis had significantly increased the acute apoptotic response to a

genotoxic carcinogen (AARGC) in rat colon [158], Le Leu et al confirmed his protective action
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against CRC in rats model, supporting the importance of this synbiotic in cancer prevention. The
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mechanism(s) of actions that sustain(s) apoptosis with this symbiotic remain(s) unclear, but authors
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supposed the presence of immunomodulating properties derived by the interaction between B.lactis

and RS resulting in butyrate production [156]. A recent randomized, double-blind, placebo-

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controlled study on humans affected by colorectal polyps (no CRC) or CRC [157] highlighted how
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the dietary symbiotic, B. lactis, L. Rhamnosus and oligofructose-enriched inulin, was able to reduce

colon cell proliferation and cell DNA-damage. However, the two groups responded differently to

the synbiotic, maybe because of an increased resistance to qualitative and quantitative changes of

cancer patients intestinal microflora [157].

In addition, the effects of Bifidobacteria are not limited to the gastrointestinal tract. In fact, a

possible therapeutic role of such probiotic has also been supposed in the treatment of respiratory

Page 14 of 36
pathologies [112-115], psoriasis, chronic fatigue syndrome [110], depression [132, 133] and in the

deterioration of cognitive functions [136].

CONCLUSIONS

This review shows how Actinobacteria phylum, despite it represents a minority group of

commensal bacteria, plays a pivotal role in the development and maintenance of gut homeostasis

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(Figure 1). Its involvement has been supposed in the modulation of gut permeability, immune

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system, metabolism and gut-brain axis. An unbalanced abundance has been evidenced in several

pathological conditions. For this reason, the interest in the use of Actinobacteria, especially of

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Bifidobacteria family, as probiotic is constantly increasing. This group may represent a significant

part of the next-generation of probiotics with a potential efficacy both in gut diseases and extra-

intestinal disorders (Table 1).

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However, further studies are needed to deeply understand the interaction between Actinobacteria

and the host and consequently their therapeutic implications in the prevention and treatment of
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several systemic disorders with the end goal of promoting gut health.
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2. Leser TD, Molbach L: Better living through microbial action: the benefits of the mammalian

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FIGURE LEGEND

Figure 1

Actinobacteria phylum groups four main families: Bifidobacteria, Propionibacteria, Corynebacteria,

Stremptomyces. Actinobacteria exert crucial physiological functions in the gut. FOS: fructo-

oligosaccharides, GOS: galacto-oligosaccharides, XOS: xylo-oligosaccharides, BDNF: brain-

derived neurotrophic factor.

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Table 1

Potential modulatory and therapeutic role of Actinobacteria in intestinal and extra-intestinal

diseases.

INTESTINAL DISEASES EXTRA-INTESTINAL DISEASES

UC: prevention of pouchitis relapse, NASH

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maintenance of remission and

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treatment of mild-to-moderate UC
IBS RESPIRATORY PATHOLOGIES

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CONSTIPATION PSORIASIS

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ANTIBIOTIC-ASSOCIATED CHRONIC FATIGUE SYNDROME
DIARHEA
NEC DEPRESSION
COLORECTAL CANCER
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COGNITIVE DETERIORATION

RCU/ UC: ulcerative colitis, IBS: irritable bowel syndrome, NEC: necrotizing entero-colitis,
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NASH: non-alcoholic steato-hepatitis.
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CONFLICT OF INTEREST

Authors declare no conflict of interest.

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Figure 1

i
cr
PHYSIOLOGICAL FUNCTIONS

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Immunological function:
Gut Barrier: -maintenance of gut permeability
-production of acetate
-down-regulation of inflammation by

M
-production of lactate that is used by
production of IL-4 and IL-13
“lactate-utilizer” for the production of
-induction of regulatory T-cells
butyrate

ed
-regulation of Th-2 immunoresponse
-maintenance of gut permeability

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Metabolism: Gut-Brain Axis:
-production of acetate -potential anti-depressant effects with the
elevation of tryptophan levels
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-biodegradation of resistant starch with

production of FOS, GOS, XOS, inulin, -regulation of mediators involved in
arabinoxilan synaptic plasticity like BDNF, synapsin,
-formation of conjugated linoleic acids Actinobacteria syntaxin
-modulation of hippocampal molecules such
as CD68 and CD11b
Bifidobacteria Propionibacteria

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Corynebacteria Streptomyces