Bifidobacterium animalis subsp. lactis modulates early-life immune response and gut metabolism

Abstract

Background: The maturation of the immune system is critical during early life, as it involves the differentiation, maturation, and establishment of immune tolerance of immune cells. This process is influenced not only by genetic factors but also by environmental factors, particularly the symbiotic microbiota. Bifidobacterium animalis subsp. lactis (BB-12), originally found in dairy products, is widely used in infant formula and dietary supplements. However, its role and mechanisms in immune development during early life remain unclear.

Methods: Using GF mice as the experimental model, B. animalis subsp. lactis BB-12 was administered via gavage during early life. In the juvenile stage, changes in T-cell subsets in the spleen, thymus, and gut intraepithelial lymphocytes (IEL) were assessed using spectral flow cytometry. Additionally, targeted metabolomics analysis of tryptophan metabolism and short-chain fatty acid pathways in colonic tissue was conducted to explore how B. animalis subsp. lactis BB-12 influences the immune system through gut microbiota metabolism.

Results: BB-12 effectively modulates the gut immune microenvironment, leading to beneficial changes in T-cell subsets in key immune tissues such as the spleen, thymus, and gut IELs. Metabolomics analysis further supports these findings by showing that BB-12 intervention greatly increased the production of tryptophan derivatives and acetic acid in the colon of GF mice.

Conclusion: The findings provide theoretical evidence for the role of B. animalis subsp. lactis in immune system development and support its application in dietary supplements, suggesting potential as a component for infant immune health and in preventing immune-related diseases.

Keywords

Bifidobacterium animalis subsp. lactis / germ-free model / gut microbiota / immune homeostasis / metabolism

References

[1] Henrick BM, Rodriguez L, Lakshmikanth T, et al. Bifidobacteria-mediated immune system imprinting early in life. Cell. 2021; 184(15): 3884-3898.

[2] Sanders ME, Merenstein DJ, Reid G, Gibson GR, Rastall RA. Probiotics and prebiotics in intestinal health and disease: from biology to the clinic. Nat Rev Gastroenterol Hepatol. 2019; 16: 605-616.

[3] Xu D, Zhou S, Liu Y, Scott AL, Yang J, Wan F. Complement in breast milk modifies offspring gut microbiota to promote infant health. Cell. 2024; 187(3): 750-763.

[4] Laursen MF, Sakanaka M, von Burg N, et al. Bifidobacterium species associated with breastfeeding produce aromatic lactic acids in the infant gut. Nat Microbiol. 2021; 6: 1367-1382.

[5] Button JE, Cosetta CM, Reens AL, et al. Precision modulation of dysbiotic adult microbiomes with a human-milk-derived synbiotic reshapes gut microbial composition and metabolites. Cell Host Microbe. 2023; 31(9): 1523-1538.

[6] Button JE, Autran CA, Reens AL, et al. Dosing a synbiotic of human milk oligosaccharides and B. Infantis leads to reversible engraftment in healthy adult microbiomes without antibiotics. Cell Host Microbe. 2022; 30: 712-725.e717.

[7] Victora CG, Bahl R, Barros AJ, et al. Breastfeeding in the 21st century: epidemiology, mechanisms, and lifelong effect. Lancet (London, England). 2016; 387: 475-490.

[8] Atyeo C, Alter G. The multifaceted roles of breast milk antibodies. Cell. 2021; 184: 1486-1499.

[9] Jungersen M, Wind A, Johansen E, Christensen J, Stuer-Lauridsen B, Eskesen D. The science behind the probiotic strain Bifidobacterium animalis subsp. lactis BB-12(®). Microorganisms. 2014; 2: 92-110.

[10] Reuter G. The lactobacillus and Bifidobacterium microflora of the human intestine: composition and succession. Curr Issues Intest Microbiol. 2001; 2: 43-53.

[11] Nocerino R, de Filippis F, Cecere G, et al. The therapeutic efficacy of Bifidobacterium animalis subsp. lactis BB-12(®) in infant colic: a randomised, double blind, placebo-controlled trial. Aliment Pharmacol Ther. 2020; 51: 110-120.

[12] Chouraqui JP, Van Egroo LD, Fichot MC. Acidified milk formula supplemented with bifidobacterium lactis: impact on infant diarrhea in residential care settings. J Pediatr Gastroenterol Nutr. 2004; 38: 288-292.

[13] Donald K, Finlay BB. Early-life interactions between the microbiota and immune system: impact on immune system development and atopic disease. Nat Rev Immunol. 2023; 23: 735-748.

[14] Bach JF. The hygiene hypothesis in autoimmunity: the role of pathogens and commensals. Nat Rev Immunol. 2018; 18: 105-120.

[15] Gaisford W, Cooke A. Can infections protect against autoimmunity? Curr Opin Rheumatol. 2009; 21: 391-396.

[16] Rizzardini G, Eskesen D, Calder PC, Capetti A, Jespersen L, Clerici M. Evaluation of the immune benefits of two probiotic strains Bifidobacterium animalis ssp. lactis, BB-12® and lactobacillus paracasei ssp. paracasei, L. casei 431® in an influenza vaccination model: a randomised, double-blind, placebo-controlled study. Br J Nutr. 2012; 107: 876-884.

[17] Taipale T, Pienihäkkinen K, Isolauri E, et al. Bifidobacterium animalis subsp. lactis BB-12 in reducing the risk of infections in infancy. Br J Nutr. 2011; 105(3): 409-416.

[18] Van Rossum T, Haiß A, Knoll RL, et al. Bifidobacterium and lactobacillus probiotics and gut dysbiosis in preterm infants: the PRIMAL randomized clinical trial. JAMA Pediatr. 2024; 178: 985-995.

[19] Wang X, Tian R, Liang C, et al. Biomimetic nanoplatform with microbiome modulation and antioxidant functions ameliorating insulin resistance and pancreatic β-cell dysfunction for T2DM management. Biomaterials. 2025; 313: 122804.

[20] Chen H, Wan X, He Q, et al. Single-cell RNA sequencing reveals cellular dynamics and therapeutic effects of astragaloside IV in slow transit constipation. Biomolecules & Biomedicine. 2024; 24: 871-887.

[21] Mebius RE, Kraal G. Structure and function of the spleen. Nat Rev Immunol. 2005; 5: 606-616.

[22] Fike AJ, Nguyen LT, Kumova OK, Carey AJ. Characterization of CD31 expression on murine and human neonatal T lymphocytes during development and activation. Pediatr Res. 2017; 82: 133-140.

[23] Natalini A, Simonetti S, Favaretto G, et al. OMIP-079: cell cycle of CD4(+) and CD8(+) naïve/memory T cell subsets, and of Treg cells from mouse spleen. Cytometry Part A. 2021; 99: 1171-1175.

[24] Nakajima Y, Chamoto K, Oura T, Honjo T. Critical role of the CD44(low)CD62L(low) CD8(+) T cell subset in restoring antitumor immunity in aged mice. Proc Natl Acad Sci U S A. 2021; 118.

[25] Youngblood B, Hale JS, Ahmed R. T-cell memory differentiation: insights from transcriptional signatures and epigenetics. Immunology. 2013; 139: 277-284.

[26] Cibrián D, Sánchez-Madrid F. CD69: from activation marker to metabolic gatekeeper. Eur J Immunol. 2017; 47: 946-953.

[27] Ivanov II, Tuganbaev T, Skelly AN, Honda K. T cell responses to the microbiota. Annu Rev Immunol. 2022; 40: 559-587.

[28] Sefik E, Geva-Zatorsky N, Oh S, et al. MUCOSAL IMMUNOLOGY. Individual intestinal symbionts induce a distinct population of RORγ⁺ regulatory T cells. Science (New York, NY). 2015; 349: 993-997.

[29] Ashby KM, Hogquist KA. A guide to thymic selection of T cells. Nat Rev Immunol. 2024; 24: 103-117.

[30] Nunes-Alves C, Nobrega C, Behar SM, Correia-Neves M. Tolerance has its limits: how the thymus copes with infection. Trends Immunol. 2013; 34: 502-510.

[31] Laouar A. Maternal leukocytes and infant immune programming during breastfeeding. Trends Immunol. 2020; 41: 225-239.

[32] Olivares-Villagómez D, Van Kaer L. Intestinal intraepithelial lymphocytes: sentinels of the mucosal barrier. Trends Immunol. 2018; 39: 264-275.

[33] Cheroutre H, Lambolez F, Mucida D. The light and dark sides of intestinal intraepithelial lymphocytes. Nat Rev Immunol. 2011; 11: 445-456.

[34] Agus A, Planchais J, Sokol H. Gut microbiota regulation of tryptophan metabolism in health and disease. Cell Host Microbe. 2018; 23: 716-724.

[35] Lavelle A, Sokol H. Gut microbiota-derived metabolites as key actors in inflammatory bowel disease. Nat Rev Gastroenterol Hepatol. 2020; 17: 223-237.

[36] Cervenka I, Agudelo LZ, Ruas JL. Kynurenines: Tryptophan's metabolites in exercise, inflammation, and mental health. Science (New York, NY). 2017; 357(6349): eaaf9794.

[37] Miao H, Wang YN, Yu XY, et al. Lactobacillus species ameliorate membranous nephropathy through inhibiting the aryl hydrocarbon receptor pathway via tryptophan-produced indole metabolites. Br J Pharmacol. 2024; 181(1): 162-179.

[38] Tian P, Chen Y, Zhu H, et al. Bifidobacterium breve CCFM1025 attenuates major depression disorder via regulating gut microbiome and tryptophan metabolism: a randomized clinical trial. Brain Behav Immun. 2022; 100: 233-241.

[39] Wu H, Singer J, Kwan TK, et al. Gut microbial metabolites induce donor-specific tolerance of kidney allografts through induction of T regulatory cells by short-chain fatty acids. J Am Soc Nephrol. 2020; 31: 1445-1461.

[40] Geuking MB, McCoy KD, Macpherson AJ. Metabolites from intestinal microbes shape Treg. Cell Res. 2013; 23: 1339-1340.

[41] Wu H, Herr D, MacIver NJ, Rathmell JC, Gerriets VA. CD4 T cells differentially express cellular machinery for serotonin signaling, synthesis, and metabolism. Int Immunopharmacol. 2020; 88: 106922.

[42] Schmidt RM, Pilmann Laursen R, Bruun S, et al. Probiotics in late infancy reduce the incidence of eczema: a randomized controlled trial. Pediatr Allergy Immunol. 2019; 30: 335-340.

[43] Lee Y, Ba Z, Roberts RF, et al. Effects of Bifidobacterium animalis subsp. lactis BB-12(®) on the lipid/lipoprotein profile and short chain fatty acids in healthy young adults: a randomized controlled trial. Nutr J. 2017; 16: 39.

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2025 The Author(s). Animal Models and Experimental Medicine published by John Wiley & Sons Australia, Ltd on behalf of The Chinese Association for Laboratory Animal Sciences.