Immunomodulatory Effects of Probiotic Lactobacillus Strains on Mucosal Immunity
DOI:
https://doi.org/10.5281/zenodo.22687982Keywords:
probiotics, mucosal immunity, secretory IgA, regulatory T cells, gut microbiota, LactobacillusAbstract
Probiotic Lactobacillus strains are marketed on the assumption that immunomodulatory benefit is a genus-level
property, yet strain-to-strain variation in mechanistic studies is substantial enough to make that assumption difficult
to defend without direct comparison under matched conditions. We compared four Lactobacillus strains commonly
used in commercial probiotic formulations -- L. rhamnosus GG, L. plantarum WCFS1, L. reuteri DSM 17938 and L.
casei Shirota -- for their effects on mucosal immune parameters in specific-pathogen-free C57BL/6 mice following
21 days of daily oral gavage, using a vehicle-only gavage group as control. Faecal and intestinal secretory IgA
concentration increased significantly relative to control for three of the four strains (L. rhamnosus GG, L. plantarum
WCFS1 and L. reuteri DSM 17938; all P<0.01), but not for L. casei Shirota (P=0.34), despite comparable
colonisation levels across all four strains as assessed by strain-specific qPCR of faecal DNA. Flow cytometric
analysis of mesenteric lymph node and lamina propria lymphocytes found a significant increase in Foxp3+
regulatory T cell frequency for L. reuteri DSM 17938 alone (1.8-fold over control, P=0.002), while L. rhamnosus
GG and L. plantarum WCFS1 instead increased the frequency of RORgt+ Th17 cells without a corresponding
regulatory T cell shift. Cytokine profiling of colonic explant culture supernatant showed a broadly pro-regulatory
pattern (elevated IL-10, reduced IL-6 and TNF-alpha) specific to the L. reuteri DSM 17938 group, while the L.
rhamnosus GG and L. plantarum WCFS1 groups showed elevated IL-17A and IL-22 without a corresponding
change in IL-10. These results demonstrate that closely related Lactobacillus strains produce qualitatively distinct,
not merely quantitatively different, mucosal immune signatures, and argue against treating strain-level
immunomodulatory claims as interchangeable within the genus, a distinction with direct relevance to how probiotic
products are formulated, marketed and selected for specific clinical indications.
References
Hill C, Guarner F, Reid G, Gibson GR, Merenstein DJ, Pot B, Morelli L, Canani RB, Flint HJ, Salminen S (2014) Expert consensus document: the International Scientific Association for Probiotics and Prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nature Reviews Gastroenterology & Hepatology 11(8):506-514. https://doi.org/10.1038/nrgastro.2014.66
Sanders ME, Merenstein DJ, Reid G, Gibson GR, Rastall RA (2019) Probiotics and prebiotics in intestinal health and disease: from biology to the clinic. Nature Reviews Gastroenterology & Hepatology 16(10):605-616. https://doi.org/10.1038/s41575-019-0173-3
Suez J, Zmora N, Segal E, Elinav E (2019) The pros, cons, and many unknowns of probiotics. Nature Medicine 25(5):716-729. https://doi.org/10.1038/s41591-019-0439-x
Segers ME, Lebeer S (2014) Towards a better understanding of Lactobacillus rhamnosus GG - host interactions. Microbial Cell Factories 13(Suppl 1):S7. https://doi.org/10.1186/1475-2859-13-S1-S7
Kwon HK, Lee CG, So JS, Chae CS, Hwang JS, Sahoo A, Nam JH, Rhee JH, Hwang KC, Im SH (2010) Generation of regulatory dendritic cells and CD4+Foxp3+ T cells by a combination of Lactobacillus rhamnosus and low-dose transforming growth factor-beta. PNAS 107(5):2159-2164. https://doi.org/10.1073/pnas.0904055107
Liu Y, Fatheree NY, Mangalat N, Rhoads JM (2010) Human-derived probiotic Lactobacillus reuteri strains differentially reduce intestinal inflammation. American Journal of Physiology-Gastrointestinal and Liver Physiology 299(5):G1087-G1096. https://doi.org/10.1152/ajpgi.00124.2010
He B, Hoang TK, Wang T, Ferris M, Taylor CM, Tian X, Luo M, Tran DQ, Zhou J, Tatevian N (2017) Resetting microbiota by Lactobacillus reuteri inhibits T reg deficiency-induced autoimmunity via adenosine A2A receptors. Journal of Experimental Medicine 214(1):107-123. https://doi.org/10.1084/jem.20160961
Macpherson AJ, Uhr T (2004) Induction of protective IgA by intestinal dendritic cells carrying commensal bacteria. Science 303(5664):1662-1665. https://doi.org/10.1126/science.1091334
Pabst O, Slack E (2020) IgA and the intestinal microbiota: the importance of being specific. Mucosal Immunology 13(1):12-21. https://doi.org/10.1038/s41385-019-0227-4
Kabat AM, Srinivasan N, Maloy KJ (2014) Modulation of immune development and function by intestinal microbiota. Trends in Immunology 35(11):507-517. https://doi.org/10.1016/j.it.2014.07.010
Bermudez-Brito M, Plaza-Diaz J, Munoz-Quezada S, Gomez-Llorente C, Gil A (2012) Probiotic mechanisms of action. Annals of Nutrition and Metabolism 61(2):160-174. https://doi.org/10.1159/000342079
Lebeer S, Vanderleyden J, De Keersmaecker SC (2010) Host interactions of probiotic bacterial surface molecules: comparison with commensals and pathogens. Nature Reviews Microbiology 8(3):171-184. https://doi.org/10.1038/nrmicro2297
Atarashi K, Tanoue T, Shima T, Imaoka A, Kuwahara T, Momose Y, Cheng G, Yamasaki S, Saito T, Ohba Y (2011) Induction of colonic regulatory T cells by indigenous Clostridium species. Science 331(6015):337-341. https://doi.org/10.1126/science.1198469
Ivanov II, Frutos RD, Manel N, Yoshinaga K, Rifkin DB, Sartor RB, Finlay BB, Littman DR (2008) Specific microbiota direct the differentiation of IL-17-producing T-helper cells in the mucosa of the small intestine. Cell Host & Microbe 4(4):337-349. https://doi.org/10.1016/j.chom.2008.09.009
Vandenplas Y, Huys G, Daube G (2015) Probiotics: an update. Jornal de Pediatria 91(1):6-21. https://doi.org/10.1016/j.jped.2014.08.005
Sun J, Chang EB (2014) Exploring gut microbes in human health and disease: pushing the envelope. Genes & Diseases 1(2):132-139. https://doi.org/10.1016/j.gendis.2014.08.001
Round JL, Mazmanian SK (2009) The gut microbiota shapes intestinal immune responses during health and disease. Nature Reviews Immunology 9(5):313-323. https://doi.org/10.1038/nri2515
Belkaid Y, Hand TW (2014) Role of the microbiota in immunity and inflammation. Cell 157(1):121-141. https://doi.org/10.1016/j.cell.2014.03.011
Zeuthen LH, Fink LN, Frokiaer H (2008) Toll-like receptor 2 and nucleotide-binding oligomerization domain-2 play divergent roles in the recognition of gut-derived lactobacilli and bifidobacteria in dendritic cells. Immunology 124(4):489-502. https://doi.org/10.1111/j.1365-2567.2007.02800.x
Konieczna P, Groeger D, Ziegler M, Frei R, Ferstl R, Shanahan F, Quigley EM, Kiely B, Akdis CA, O'Mahony L (2012) Bifidobacterium infantis 35624 administration induces Foxp3 T regulatory cells in human peripheral blood. Gut 61(3):354-366. https://doi.org/10.1136/gutjnl-2011-300936
Yan F, Polk DB (2011) Probiotics and immune health. Current Opinion in Gastroenterology 27(6):496-501. https://doi.org/10.1097/MOG.0b013e32834baa4d
Kleerebezem M, Boekhorst J, van Kranenburg R, Molenaar D, Kuipers OP, Leer R, Tarchini R, Peters SA, Sandbrink HM, Fiers MW (2003) Complete genome sequence of Lactobacillus plantarum WCFS1. PNAS 100(4):1990-1995. https://doi.org/10.1073/pnas.0337704100
van Baarlen P, Troost F, van der Meer C, Hooiveld G, Boekschoten M, Brummer RJ, Kleerebezem M (2011) Human mucosal in vivo transcriptome responses to three lactobacilli indicate how probiotics may modulate human cellular pathways. PNAS 108(Suppl 1):4562-4569. https://doi.org/10.1073/pnas.1000079107
Goldenberg JZ, Yap C, Lytvyn L, Lo CK, Beardsley J, Mertz D, Johnston BC (2017) Probiotics for the prevention of Clostridium difficile-associated diarrhea in adults and children. Cochrane Database of Systematic Reviews 12:CD006095. https://doi.org/10.1002/14651858.CD006095.pub4
Basturk A, Artan R, Yilmaz A (2016) Efficacy of synbiotic, probiotic, and prebiotic treatments for irritable bowel syndrome in children: a randomized controlled trial. Turkish Journal of Gastroenterology 27(5):439-443. https://doi.org/10.5152/tjg.2016.16301
Rautava S, Kalliomaki M, Isolauri E (2002) Probiotics during pregnancy and breast-feeding might confer immunomodulatory protection against atopic disease in the infant. Journal of Allergy and Clinical Immunology 109(1):119-121. https://doi.org/10.1067/mai.2002.120273
Wildt S, Munck LK, Vinter-Jensen L, Hanse BF, Nordgaard-Lassen I, Christensen S, Avnstroem S, Rasmussen SN, Rumessen JJ (2011) Probiotic treatment of collagenous colitis: a randomized, double-blind, placebo-controlled trial with Lactobacillus acidophilus and Bifidobacterium animalis subsp. lactis. Inflammatory Bowel Diseases 17(1):125-132. https://doi.org/10.1002/ibd.21339
Fujimura KE, Sitarik AR, Havstad S, Lin DL, Levan S, Fadrosh D, Panzer AR, LaMere B, Rackaityte E, Lukacs NW (2016) Neonatal gut microbiota associates with childhood multisensitized atopy and T cell differentiation. Nature Medicine 22(10):1187-1191. https://doi.org/10.1038/nm.4176
Mazmanian SK, Liu CH, Tzianabos AO, Kasper DL (2005) An immunomodulatory molecule of symbiotic bacteria directs maturation of the host immune system. Cell 122(1):107-118. https://doi.org/10.1016/j.cell.2005.05.007
Chung H, Pamp SJ, Hill JA, Surana NK, Edelman SM, Troy EB, Reading NC, Villablanca EJ, Wang S, Mora JR (2012) Gut immune maturation depends on colonization with a host-specific microbiota. Cell 149(7):1578-1593. https://doi.org/10.1016/j.cell.2012.04.037
Smits HH, Engering A, van der Kleij D, de Jong EC, Schipper K, van Capel TM, Zaat BA, Yazdanbakhsh M, Wierenga EA, van Kooyk Y (2005) Selective probiotic bacteria induce IL-10-producing regulatory T cells in vitro by modulating dendritic cell function through dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin. Journal of Allergy and Clinical Immunology 115(6):1260-1267. https://doi.org/10.1016/j.jaci.2005.03.036
Foligne B, Zoumpopoulou G, Dewulf J, Ben Younes A, Chareyre F, Sirard JC, Pot B, Grangette C (2007) A key role of dendritic cells in probiotic functionality. PLOS ONE 2(3):e313. https://doi.org/10.1371/journal.pone.0000313
Rescigno M (2011) The intestinal epithelial barrier in the control of homeostasis and immunity. Trends in Immunology 32(6):256-264. https://doi.org/10.1016/j.it.2011.04.003
Ouwehand AC, Salminen S, Isolauri E (2002) Probiotics: an overview of beneficial effects. Antonie van Leeuwenhoek 82(1-4):279-289. https://doi.org/10.1023/A:1020620607611
Kailasapathy K, Chin J (2000) Survival and therapeutic potential of probiotic organisms with reference to Lactobacillus acidophilus and Bifidobacterium spp. Immunology and Cell Biology 78(1):80-88. https://doi.org/10.1046/j.1440-1711.2000.00886.x
Gill HS, Rutherfurd KJ, Cross ML, Gopal PK (2001) Enhancement of immunity in the elderly by dietary supplementation with the probiotic Bifidobacterium lactis HN019. American Journal of Clinical Nutrition 74(6):833-839. https://doi.org/10.1093/ajcn/74.6.833
Weiss G, Rasmussen S, Zeuthen LH, Nielsen BN, Jarmer H, Jespersen L, Frokiaer H (2010) Lactobacillus acidophilus induces virus immune defence genes in murine dendritic cells by a Toll-like receptor-2-dependent mechanism. Immunology 131(2):268-281. https://doi.org/10.1111/j.1365-2567.2010.03301.x
Ghadimi D, Folster-Holst R, de Vrese M, Winkler P, Heller KJ, Schrezenmeir J (2008) Effects of probiotic bacteria and their genomic DNA on TH1/TH2-cytokine production by peripheral blood mononuclear cells (PBMCs) of healthy and allergic subjects. Immunobiology 213(8):677-692. https://doi.org/10.1016/j.imbio.2008.02.001
Meijerink M, van Hemert S, Taverne N, Wels M, de Vos P, Bron PA, Savelkoul HF, van Bilsen J, Kleerebezem M, Wells JM (2010) Identification of genetic loci in Lactobacillus plantarum that modulate the immune response of dendritic cells using comparative genome hybridization. PLOS ONE 5(5):e10632. https://doi.org/10.1371/journal.pone.0010632
Wells JM (2011) Immunomodulatory mechanisms of lactobacilli. Microbial Cell Factories 10(Suppl 1):S17. https://doi.org/10.1186/1475-2859-10-S1-S17
Bron PA, Kleerebezem M (2018) Lactic acid bacteria for delivery of endogenous or engineered therapeutic molecules. Frontiers in Microbiology 9:1821. https://doi.org/10.3389/fmicb.2018.01821
Grimm V, Radulovic K, Riedel CU (2015) Colonization of C57BL/6 mice by a potential probiotic Bifidobacterium bifidum strain under germ-free and specific pathogen-free conditions and during experimental colitis. PLOS ONE 10(10):e0139935. https://doi.org/10.1371/journal.pone.0139935
Cebra JJ (1999) Influences of microbiota on intestinal immune system development. American Journal of Clinical Nutrition 69(5):1046S-1051S. https://doi.org/10.1093/ajcn/69.5.1046s
Cash HL, Whitham CV, Behrendt CL, Hooper LV (2006) Symbiotic bacteria direct expression of an intestinal bactericidal lectin. Science 313(5790):1126-1130. https://doi.org/10.1126/science.1127119
Kwon HK, Kim GC, Kim Y, Hwang W, Jash A, Sahoo A, Kim JE, Nam JH, Im SH (2013) Amelioration of experimental autoimmune encephalomyelitis by probiotic mixture is mediated by a shift in T helper cell immune response. Clinical Immunology 146(3):217-227. https://doi.org/10.1016/j.clim.2013.01.001
Zocco MA, Ainora ME, Gasbarrini G, Gasbarrini A (2007) Bacteroides thetaiotaomicron in the gut: molecular aspects of their interaction. Digestive and Liver Disease 39(8):707-712. https://doi.org/10.1016/j.dld.2007.04.003
Ashraf R, Shah NP (2014) Immune system stimulation by probiotic microorganisms. Critical Reviews in Food Science and Nutrition 54(7):938-956. https://doi.org/10.1080/10408398.2011.619671
Delcenserie V, Martel D, Lamoureux M, Amiot J, Boutin Y, Roy D (2008) Immunomodulatory effects of probiotics in the intestinal tract. Current Issues in Molecular Biology 10(1-2):37-54. https://doi.org/10.21775/cimb.010.037
Jang SO, Kim HJ, Kim YJ, Kang MJ, Kwon JW, Seo JH, Kim HY, Kim BJ, Yu J, Hong SJ (2012) Asthma prevention by Lactobacillus rhamnosus in a mouse model is associated with CD4+CD25+Foxp3+ T cells. Allergy, Asthma & Immunology Research 4(3):150-156. https://doi.org/10.4168/aair.2012.4.3.150
