Influence of early gut microbiota on the maturation of childhood mucosal and systemic immune responses.
BACKGROUND
Among sensitized infants, those with high, as compared with low levels, of salivary secretory IgA (SIgA) are less likely to develop allergic symptoms. Also, early colonization with certain gut microbiota, e.g. Lactobacilli and Bifidobacterium species, might be associated with less allergy development. Although animal and in vitro studies emphasize the role of the commensal gut microbiota in the development of the immune system, the influence of the gut microbiota on immune development in infants is unclear.


OBJECTIVE
To assess whether early colonization with certain gut microbiota species associates with mucosal and systemic immune responses i.e. salivary SIgA and the spontaneous Toll-like receptor (TLR) 2 and TLR4 mRNA expression and lipopolysaccharide (LPS)-induced cytokine/chemokine responses in peripheral blood mononuclear cells (PBMCs).


METHODS
Fecal samples were collected at 1 week, 1 month and 2 months after birth from 64 Swedish infants, followed prospectively up to 5 years of age. Bacterial DNA was analysed with real-time PCR using primers binding to Clostridium difficile, four species of bifidobacteria, two lactobacilli groups and Bacteroides fragilis. Saliva was collected at age 6 and 12 months and at 2 and 5 years and SIgA was measured with ELISA. The PBMCs, collected 12 months after birth, were analysed for TLR2 and TLR4 mRNA expression with real-time PCR. Further, the PBMCs were stimulated with LPS, and cytokine/chemokine responses were measured with Luminex.


RESULTS
The number of Bifidobacterium species in the early fecal samples correlated significantly with the total levels of salivary SIgA at 6 months. Early colonization with Bifidobacterium species, lactobacilli groups or C. difficile did not influence TLR2 and TLR4 expression in PBMCs. However, PBMCs from infants colonized early with high amounts of Bacteroides fragilis expressed lower levels of TLR4 mRNA spontaneously. Furthermore, LPS-induced production of inflammatory cytokines and chemokines, e.g. IL-6 and CCL4 (MIP-1 beta), was inversely correlated to the relative amounts of Bacteroides fragilis in the early fecal samples.


CONCLUSIONS
Bifidobacterial diversity may enhance the maturation of the mucosal SIgA system and early intense colonization with Bacteroides fragilis might down-regulate LPS responsiveness in infancy.
Sjögren
Y M
YM
Department of Immunology, Wenner-Gren Institute, Stockholm University, Stockholm, Sweden. ylva.sjogren@imun.su.se
Tomicic
S
S
Lundberg
A
A
Böttcher
M F
MF
Björkstén
B
B
Sverremark-Ekström
E
E
Jenmalm
M C
MC
D001441:Bacteroides fragilis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D001644:Bifidobacterium / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D054407:Chemokine CCL4 / Q000378:metabolism; D002675:Child, Preschool; D016360:Clostridioides difficile / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology; D005260:Female; D015870:Gene Expression / Q000235:genetics / Q000276:immunology; D006801:Humans; D007107:Immune System / Q000254:growth & development* / Q000276:immunology; D007109:Immunity / Q000276:immunology*; D018928:Immunity, Mucosal / Q000276:immunology*; D007071:Immunoglobulin A, Secretory / Q000276:immunology; D007223:Infant; D007231:Infant, Newborn; D007371:Interferon-gamma / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D007378:Interleukins / Q000378:metabolism; D007422:Intestines / Q000276:immunology / Q000382:microbiology*; D007778:Lactobacillus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008137:Longitudinal Studies; D008297:Male; D054892:Metagenome / Q000276:immunology*; D010835:Phytohemagglutinins / Q000494:pharmacology; D012463:Saliva / Q000276:immunology; D051197:Toll-Like Receptor 4 / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D054407:Chemokine CCL4; C508600:IL6 protein, human; D007071:Immunoglobulin A, Secretory; D015850:Interleukin-6; D007378:Interleukins; D008070:Lipopolysaccharides; D010835:Phytohemagglutinins; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma