Gut immune homeostasis: the immunomodulatory role of Bacillus clausii, from basic to clinical evidence

Figures & data

Figure 1. Proiotics modulate the gut immune system in a variety of ways, including modifying the balance between Th1/Th2/Th17 and Treg cells, inducing the production of sIgA that inhibits adhesion and motility of pathogens, producing antimicrobial peptides such as bacteriocins and defensins, producing proteases that counter enterotoxins, competitive exclusion from mucosal adhesion sites, interaction with gut epithelial cells via cell-surface structures and increased synthesis of tight junction proteins, mucins, short-chain fatty acids, digestive enzymes, and chemicals with systemic effects, such as cortisol and tryptophan (reviewed in [7,8].

Figure 2. Exposure to different subsets of microbes due to different environments in the first 1000 days of life can lead to normal or pathological imprinting with long-term consequences on health. Caesarean section and formula milk contribute to dysbiosis in early life. Other factors such as diet and consumption of antibiotics can also lead to imbalances in the gut microbiota, ultimately contributing to chronic inflammatory disease later in life [19]. Adapted from [17].

Figure 3. Probiotics have shown promise in a variety of conditions affected by an imbalanced gut microbiota. Apart from gut-related diseases such as diarrhea, enterocolitis, IBS, IBD, and food allergies, the gut microbiota also influence diseases of the central nervous system, skin, liver, and lungs. Vaccine response and metabolic diseases are also affected by gut microbiota composition [1,4,56–75].

Table 1. Immunomodulatory effects of Bacillus probiotics

Type of study	Probiotic species/strains (dose used in clinical studies)	Property	Mechanism	Evidence of action	Reference
Clinical (RCT)	B. subtilis CU1 (2 x 10^9 spores/day)	Antimicrobial activity	Produce antibiotics (amicoumacins)	Reduce the number of respiratory infections, increased salivary and fecal levels of sIgA	[89]
	B. coagulans GBI-30, 6086 (1 x 10^9 CFU/day)	Alter the gut microbiota composition	Increase numbers of beneficial bacterial species	Increased numbers of Faecalibacterium prausnitzii, Clostridium lituseburense, and Bacillus spp.	[90,91]
	B. coagulans LBSC (6 x 10^9 CFU/day), B. coagulans IS2 (2 x 10^9 CFU/day), B. coagulans Colinox® (dosage information not available)	Alter the gut microbiota composition	Reduce the frequency of gastrointestinal symptoms such as bloating, cramping, abdominal pain, diarrhea, constipation, and stomach rumbling.	Treat irritable bowel syndrome in adults and children	[92–94]
	B. coagulans lilac-01 (1 x 10^8 CFU/day)	Produce short-chain fatty acids	Increase fecal moisture content, lower fecal pH	Treat functional constipation	[95]
Clinical (Case report)	B. subtilis 3, B. licheniformis 31 (4 x 10^9 CFU/day)	Alter the gut microbiota composition	Inhibit the growth of enteric pathogens	Reduce the incidence of AAD, nausea, vomiting, bloating and abdominal pain during antibiotic treatment.	[96]
Preclinical (in vitro)	B. subtilis Biosporin	Antimicrobial activity	Produce antibiotics	Inhibit H. pylori, Enterococcus faecium, Campylobacter jejuni, and Shigella flaxneri	[97]
	B. coagulans BDU3	Antimicrobial activity	Produce bacteriocins	Inhibit the growth of Bacillus cereus MTCC 430, Staphylococcus aureus MTCC 3160, Enterococcus sp. MTCC 9728, Lactobacillus sp. MTCC 10093, and Micrococcus luteus MTCC 106.	[98]
	B. polyfermenticus SCD	Antimicrobial activity	Produce bacteriocin (polyfermenticin SCD)	Inhibit the growth of Bacillus spp., Listeria monocytogenes ATCC 15313, Micrococcus flavus ATCC 10240, S. aureus ATCC 25923 and KCCM 32359, and Clostridium perfringens ATCC 3624	[99]
	B. subtilis KKU213	Antimicrobial activity	Produce bacteriocin (subtilosin A)	Inhibit Bacillus cereus MC11, Listeria monocytogenes DMST23145, Micrococcus luteus MC45, Staphylococcus aureus MC13, and Enterococcus faecalis BT2 and MG30.	[100]
	Bacillus indicus HU36, B. subtilis HU58, B. coagulans SC-208, B. licheniformis and B. clausii SC-109 spores	Alter the gut microbiota composition	Increase the levels of butyrate, acetate, and propionate, increase the numbers of Bifidobacteriaceae, Lactobacillaceae, and Prevotellaceae	Maintain eubiosis	[101]
Preclinical (cell line)	B. amyloliquefaciens CCTCC M 2012280	Alter pro- and anti-inflammatory responses	Increase expression of inflammatory cell-surface markers, CD80, CD86 and MHC-II, polarize macrophages into pro-inflammatory M1 type, and increase NO production and phagocytosis by M1 macrophages	Destroy tumor cells and intracellular pathogens	[102]
Preclinical (mice)	Mix of B. subtilis and Enterococcus faecium	Alter pro- and anti-inflammatory responses	Reduce mortality, reduce levels of IL-6 and TNF-α, suppress macrophage activation to M1 type	Improve survival outcomes in sepsis	[103]
	Exopolysaccharide of B. subtilis	Alter pro- and anti-inflammatory responses	Polarize macrophages into anti-inflammatory M2 type, reduce Citrobacter rodentium-driven hyperinflammation.	Inhibit acute colitis caused by Citrobacter rodentium	[104]
	B. subtilis PB6	Alter the levels of pro- and anti-inflammatory cytokines	Induce higher expression of the anti-inflammatory cytokine, IL-10, and lower levels of the pro-inflammatory IL-12 and IFN-γ	Prevent colitis development	[105]
Preclinical (rats)	B. coagulans (strain not mentioned)	Alter the gut microbiota composition and reduce chronic inflammation	Improve clinical and biochemical parameters of rheumatoid arthritis	Treat rheumatoid arthritis	[106]

AAD, antibiotic-associated diarrhea; IFN, interferon; IL, interleukin; MHC, major histocompatibility complex.

Table 2. Modes of action identified for B. clausii from preclinical studies

Mode of action	Property of B. clausii	Reference
Alter pro- and anti-inflammatory responses	Increase expression of IL-6, IL-10, IL-12, IFN-α, and IFN-γ Decrease expression of TNF- α Inhibit release of IL-8 and IFN-β Stimulate production of intermediate levels of NO Stimulate proliferation of CD4^+ T cells	[88,113,117,119]
Antimicrobial activity	Production of clausin Production of human beta-defensin-2 and cathelicidin LL-32	[116,117]
Inhibit enterotoxins	Serine protease activity that inhibits enterotoxins produced by C. difficile	[121]
Enhance barrier function	Increase production of MUC5AC, ZO-1 Express and secrete proteins that bind plasminogen/fibronectin/mucus	[80,117,118]

IFN, interferon; IL, interleukin; MUC5AC, mucin 5AC; NO, nitric oxide; ZO-1, TNF, tumor necrosis factor; Zonula occludens protein 1.

Table 3. The effect of B. clausii on the clinical outcomes of different diseases

Condition	Strain	Outcome	Study design	Reference
Acute pediatric diarrhea	O/C, SIN, N/R, T (2 x 10^9 to 4 x 10^9 CFU/day)	Reduced duration of diarrhea	Prospective, open-label, multi-center, observational study; n = 3178	[126]
	UBBC-07 4 x 10^9 spores/day	Reduced duration of diarrhea, reduced stool frequency	Randomized, double-blind, placebo-controlled trial; n = 153	[127]
Rotavirus infection	Strain not mentioned	Reduction in general weakness, swelling and/or abdominal pain, fever, diarrhea duration, vomiting. Normalization of IgA and IgM levels to pre-infection levels.	Observational study; n = 65	[128]
	O/C, SIN, N/R, T (2 x 10^9 to 4 x 10^9 CFU/day)	Reduced duration of diarrhea, reduced stool frequency, shortened hospital stay	Meta-analysis; n = 898 from 6 studies	[124]
H. pylori treatment	O/C, SIN, N/R, T (6 x 10^9 spores/day)	Reduced diarrhea, nausea and epigastric pain	Single-center, double blind, randomized, placebo-controlled prospective study; n = 120	[87]
	O/C, SIN, N/R, T (6 x 10^9 CFU/day)	Lower incidence of diarrhea, fewer days with diarrhea	Randomized, double blind, single-center, placebo-controlled, parallel group, phase 3b study; n = 130	[129]
Necrotizing enterocolitis & late-onset sepsis	O/C, SIN, N/R, T	Significantly faster attainment of full feeds	Randomized, double-blind, placebo-controlled trial; n = 244	[132]
Respiratory tract infections	O/C, SIN, N/R, T	Reduced number and duration of infections during treatment and follow-up periods	Randomized, single-blind, multi-center, two arm parallel group study; n = 80	[136]
Allergic rhinitis	O/C, SIN, N/R, T	Decrease in pro-inflammatory cytokines, increase in anti-inflammatory cytokines	Single-blind, non-controlled study; n = 10 Randomized, double-blind, placebo-controlled study; n = 20	[137,138]

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Data availability statement

There are no datasets associated with this article.