Lactobacillus plantarum 299v probiotic supplementation in men with stable coronary artery disease suppresses systemic inflammation

Benjamin C Hofeld, Venkata K Puppala, Sudhi Tyagi, Kwang Woo Ahn, Amberly Anger, Shuang Jia, Nita H Salzman, Martin J Hessner, Michael E Widlansky, Benjamin C Hofeld, Venkata K Puppala, Sudhi Tyagi, Kwang Woo Ahn, Amberly Anger, Shuang Jia, Nita H Salzman, Martin J Hessner, Michael E Widlansky

Abstract

Recent trials demonstrate that systemic anti-inflammatory therapy reduces cardiovascular events in coronary artery disease (CAD) patients. We recently demonstrated Lactobacillus plantarum 299v (Lp299v) supplementation improved vascular endothelial function in men with stable CAD. Whether this favorable effect is in part due to anti-inflammatory action remains unknown. Testing this hypothesis, we exposed plasma obtained before and after Lp299v supplementation from these subjects to a healthy donor's PBMCs and measured differences in the PBMC transciptome, performed gene ontological analyses, and compared Lp299v-induced transcriptome changes with changes in vascular function. Daily alcohol users (DAUs) (n = 4) had a significantly different response to Lp299v and were separated from the main analyses. Non-DAUs- (n = 15) showed improved brachial flow-mediated dilation (FMD) and reduced circulating IL-8, IL-12, and leptin. 997 genes were significantly changed. I.I.com decreased (1.01 ± 0.74 vs. 0.22 ± 0.51; P < 0.0001), indicating strong anti-inflammatory effects. Pathway analyses revealed downregulation of IL-1β, interferon-stimulated pathways, and toll-like receptor signaling, and an increase in regulator T-cell (Treg) activity. Reductions in GBP1, JAK2, and TRAIL expression correlated with improved FMD. In non-DAU men with stable CAD, post-Lp299v supplementation plasma induced anti-inflammatory transcriptome changes in human PBMCs that could benefit CAD patients. Future studies should delineate changes in circulating metabolites responsible for these effects.

Conflict of interest statement

Dr. Widlansky is supported in part by HL144098 that focuses on the impact of Lp299v supplementation. Probi (Lund, Sweden), the manufacturer of Lp299v, supports HL144098 through the donation of product. Probi did not donate any product for studies reported in this manuscript. Dr. Hessner is also supported in part by DK125014 which studies the effect of Lp299v on innate inflammation in type 1 diabetes.

Figures

Figure 1
Figure 1
Effects of Lp299v supplementation on Brachial Artery FMD, ILs-8, 12, and Leptin in Non-Daily Alcohol Drinkers: Lp299v supplementation resulted in (A) improved brachial flow mediated dilation (FMD)% (3.70 ± 0.51 vs. 4.58 ± 0.61%, n = 15; P = 0.016) while IL-8 (B, 13.8 ± 2.0 vs. 10.1 ± 1.1 pg/mL, n = 15; P = 0.032), IL-12 (C, 54.6 ± 8.1 vs. 33.0 ± 7.9 pg/mL, n = 15; P = 0.042), and leptin (D, 136 ± 25 vs. 111 ± 23 ng/mL, n = 15; P = 0.002) all significantly decreased. All statistical tests paired t-tests. All values given as mean ± SE. *p < 0.05.
Figure 2
Figure 2
Post-Lp299v Plasma-Induced PBMC Signatures‒Stratified by Daily and Non-Daily Alcohol Drinkers: (A) 1,443 probe sets representing 997 unique genes were altered by at least 20% with a qval FDR < 0.2) in 15 individuals with stable CAD who did not report daily alcohol use (left) and four individuals with stable CAD who did report daily alcohol use (right). The mean for each group is indicated and each subsequent column is a subject. Data are expressed as a fold of change post- vs pre-supplementation with Lp299v. Red represents upregulated probe sets (n = 505); green represents downregulated probe sets (n = 938). (B) Well annotated transcripts associated with inflammatory and regulatory activity that showed significant change with Lp299v supplementation in these subjects.
Figure 3
Figure 3
Impact of Lp299v supplementation on the Composite Inflammatory Index (I.I.com) for Non-Daily and Daily Alcohol Drinkers. (A) Exposure to post-Lp299v supplementation plasma from non-daily alcohol using subjects drove an anti-inflammatory gene expression profile in PBMCs compared to plasma obtained prior to Lp299v supplementations (0.22 ± 0.51 vs 1.01 ± 0.74, n = 15; P < 0.0001 by paired t-test). (B) Lp299v had no significant impact on the inflammatory gene transcription expression in subjects who reported drinking alcohol daily (0.49 ± 0.78 vs 1.01 ± 1.02, n = 4; P = 0.23 by paired t-test). All values given as mean ± SD.

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