Modulation of Macrophages TLR4-Mediated Transcriptional Response by Lacticaseibacillus rhamnosus CRL1505 and Lactiplantibacillus plantarum CRL1506

<i>Lacticaseibacillus rhamnosus</i> CRL1505 and <i>Lactiplantibacillus plantarum</i> CRL1506 increase the resistance of mice to Gram-negative pathogens infections. In this work, we advanced the characterization of the CRL1505 and CRL1506 immunomodulatory properties by evaluating their effect on the Toll-like receptor 4 (TLR4)-triggered immune response in macrophages. We performed experiments in murine RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS) to evaluate the transcriptomic changes induced by lactobacilli. These in vitro experiments were complemented with in vivo studies in mice to determine the effect of CRL1505 and CRL1506 strains on Peyer's patches and peritoneal macrophages. Microarray transcriptomic studies and qPCR confirmation showed that the CRL1505 and CRL1506 strains modulated the expression of inflammatory cytokines and chemokines as well as adhesion molecules in LPS-challenged RAW macrophages, making the effect of <i>L. rhamnosus</i> CRL1505 more remarkable. Lactobacilli also modulate regulatory factors in macrophages. <i>L. plantarum</i> CRL1506 increased <i>il10</i> and <i>socs2</i> while <i>L. rhamnosus</i> CRL1505 upregulated <i>il27</i>, <i>socs1</i>, and <i>socs3</i> in RAW cells, indicating a strain-specific effect. However, in vivo, both strains induced similar effects. Peyer's patches and peritoneal macrophages from mice treated with lactobacilli produced higher levels of tumor necrosis factor (TNF)-α, interferon (IFN)-γ, interleukin (IL)-6, and colony stimulating factor (CSF)-3 after LPS stimulation. This effect would allow improved protection against pathogens. In addition, both lactobacilli equally modulated <i>socs1</i> and <i>socs2</i> expressions and IL-10 and IL-27 production in Peyer's patches macrophages and <i>socs3</i> and IL-10 in peritoneal cells. Furthermore, lactobacilli reduced the production of IL-1β, IL-12, CSF2, C-C motif chemokine ligand (CCL)-2, and CCL8 in LPS-challenged macrophages. This differential modulation of regulatory and inflammatory factors would allow minimal inflammatory-mediated tissue damage during the generation of the innate immune response. This work provides evidence that <i>L. rhamnosus</i> CRL1505 and <i>L. plantarum</i> CRL1506 modulate macrophages' TLR4-mediated immunotranscriptomic response, helping to improve protection against Gram-negative bacterial infections.