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. 2024 Sep 6;16(17):3014.
doi: 10.3390/nu16173014.

Lipoteichoic Acid from Heyndrickxia coagulans HOM5301 Modulates the Immune Response of RAW 264.7 Macrophages

Shiqi Zhang  1   2   3 , Pinglan Li  1 , Xiao Zhang  2 , Yan Ding  2 , Tingting Wang  2 , Suwon Lee  2 , Ying Xu  2 , Chongyoon Lim  2 , Nan Shang  3   4
Affiliations

Lipoteichoic Acid from Heyndrickxia coagulans HOM5301 Modulates the Immune Response of RAW 264.7 Macrophages

Shiqi Zhang et al. Nutrients. .

Abstract

Heyndrickxia coagulans (formerly Bacillus coagulans) has been increasingly utilized as an immunomodulatory probiotics. Oral administration of H. coagulans HOM5301 significantly boosted both innate and adaptive immunity in mice, particularly by increasing the phagocytic capacity of monocytes/macrophages. Lipoteichoic acid (LTA), a major microbe-associated molecular pattern (MAMP) in Gram-positive bacteria, exhibits differential immunomodulatory effects due to its structural heterogeneity. We extracted, purified, and characterized LTA from H. coagulans HOM5301. The results showed that HOM5301 LTA consists of a glycerophosphate backbone. Its molecular weight is in the range of 10-16 kDa. HOM5301 LTA induced greater productions of nitric oxide, TNFα, and IL-6 in RAW 264.7 macrophages compared to Staphylococcus aureus LTA. Comparative transcriptome and proteome analyses identified the differentially expressed genes and proteins triggered by HOM5301 LTA. KEGG analyses revealed that HOM5301 LTA transcriptionally and translationally activated macrophages through two immune-related pathways: cytokine-cytokine receptor interaction and phagosome formation. Protein-protein interaction network analysis indicated that the pro-inflammatory response elicited by HOM5301 LTA was TLR2-dependent, possibly requiring the coreceptor CD14, and is mediated via the MAPK and NF-kappaB pathways. Our results demonstrate that LTA is an important MAMP of H. coagulans HOM5301 that boosts immune responses, suggesting that HOM5301 LTA may be a promising immunoadjuvant.

Keywords: Heyndrickxia coagulans; boosting immunity; immunoadjuvant; immunomodulatory mechanism; lipoteichoic acid; macrophages.

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Conflict of interest statement

Authors Shiqi Zhang, Xiao Zhang, Yan Ding, Tingting Wang, Suwon Lee, Ying Xu and Chongyoon Lim were employed by the company Food & Biotech R&D Center, Coree Beijing Co., Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Properties of LTA isolated from H. coagulans HOM5301. (A) HPLC-SEC-RID analysis of the purified LTA. (B) MALDI-TOF mass spectrum of purified LTA (10,000–100,000 m/z). (C) FT-IR spectra of purified LTA and SaLTA. (D) 1H-NMR spectra of purified LTA and SaLTA in D2O solvent (600 MHz).
Figure 2
Figure 2
Proliferative effects of LTA on macrophages. * indicates significantly higher than the negative control (NC) (p < 0.05); △ indicates significantly lower than the NC (p < 0.05).
Figure 3
Figure 3
Effects of LTA on the production of NO (A), TNFα (B), IL-6 (C), and IL-10 (D) in RAW 264.7 cells. The different letter labels on error bars represent statistically significant differences among the tested samples (p < 0.05).
Figure 4
Figure 4
Differential expression patterns of mRNAs and proteins between the negative control and LTA-treated RAW 264.7 macrophages. (A) Venn diagram of genes co-expressed between the NC and LTA-treated groups. (B) Venn diagram of proteins co-expressed between the NC and LTA-treated groups. (C) Volcano plot of all sequenced genes from the NC and LTA-treated groups. (D) Volcano plot of all sequenced proteins from the NC and LTA-treated groups. (E) Heat map of differentially expressed genes (DEGs) between the NC and LTA-treated groups. (F) Heat map of differentially expressed proteins (DEPs) between the NC and LTA-treated groups.
Figure 5
Figure 5
Functional enrichment analysis of DEGs and DEPs based on KEGG pathway category. (A) Scatterplot of significantly enriched KEGG pathways in DEGs (p-adjust < 0.05). (B) Scatterplot of significantly enriched KEGG pathways in DEPs (p-adjust < 0.05). The color and size of the dots represent the p-adjust values and DEP numbers, respectively.
Figure 6
Figure 6
PPI network of DEPs associated with TLR signaling and cytokine–cytokine interaction pathways. The network nodes represent the DEPs, and the lines indicate associations between the linked DEPs.
Figure 7
Figure 7
Relative expression levels of the nine selected genes in the PPI network with significant differences between the negative control and LTA-treated RAW 264.7 macrophages. (A) Tlr2. (B) Cd14. (C) Myd88. (D) Il1β. (E) Cxcl2. (F) Ccl4. (G) Tnf. (H) Cd40. (I) Tollip. Quantification results were normalized using transcripts per million. * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. NC.
Figure 8
Figure 8
The expression levels of the nine selected proteins in the PPI network with significant differences between the negative control and LTA-treated RAW 264.7 cells. (A) TLR2. (B) CD14. (C) MyD88. (D) IL1β. (E) CXCL2. (F) CCL4. (G) TNF. (H) CD40. (I) TOLLIP. * p < 0.05 and *** p < 0.001 vs. NC.
Figure 9
Figure 9
Proposed mechanisms for the immunomodulatory activity of HOM5301 LTA on macrophages.
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References

    1. Liang J., Li C., Chen Z., Guo F., Dou J., Wang T., Xu Z.S. Progress of research and application of Heyndrickxia coagulans (Bacillus coagulans) as probiotic bacteria. Front. Cell Infect. Microbiol. 2024;14:1415790. doi: 10.3389/fcimb.2024.1415790. - DOI - PMC - PubMed
    1. Anaya-Loyola M.A., Enciso-Moreno J.A., Lopez-Ramos J.E., Garcia-Marin G., Orozco Alvarez M.Y., Vega-Garcia A.M., Mosqueda J., Garcia-Gutierrez D.G., Keller D., Perez-Ramirez I.F. Bacillus coagulans GBI-30, 6068 decreases upper respiratory and gastrointestinal tract symptoms in healthy Mexican scholar-aged children by modulating immune-related proteins. Food Res. Int. 2019;125:108567. doi: 10.1016/j.foodres.2019.108567. - DOI - PubMed
    1. Hoffman J.R., Hoffman M.W., Zelicha H., Gepner Y., Willoughby D.S., Feinstein U., Ostfeld I. The effect of 2 weeks of inactivated probiotic Bacillus coagulans on endocrine, inflammatory, and performance responses during self-defense training in soldiers. J. Strength Cond. Res. 2019;33:2330–2337. doi: 10.1519/JSC.0000000000003265. - DOI - PubMed
    1. Majeed M., Nagabhushanam K., Arumugam S., Majeed S., Ali F. Bacillus coagulans MTCC 5856 for the management of major depression with irritable bowel syndrome: A randomised, double-blind, placebo controlled, multi-centre, pilot clinical study. Food Nutr. Res. 2018;62:1218. doi: 10.29219/fnr.v62.1218. - DOI - PMC - PubMed
    1. Jensen G.S., Benson K.F., Carter S.G., Endres J.R. GanedenBC30 cell wall and metabolites: Anti-inflammatory and immune modulating effects in vitro. BMC Immunol. 2010;11:1–14. doi: 10.1186/1471-2172-11-15. - DOI - PMC - PubMed

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