Overcoming

Aging-Associated Poor Influenza Vaccine Responses with CpG 1018 Adjuvant

Xinliang Kang, Yibo LI, Yiwen Zhao, Xinyuan Chen

Aging is associated with diminished immune system function, which renders old people vulnerable to influenza infection and also less responsive to influenza vaccination. This study explored whether the CpG 1018 adjuvant was effective in enhancing influenza vaccine efficacy in aged mice equivalent to human beings in their late 50s to early 60s. Using the influenza pandemic 2009 H1N1 (pdm09) vaccine as a model, we found that the CpG 1018 adjuvant could significantly enhance the pdm09 vaccine-induced serum antibody titer, while the pdm09 vaccine alone failed to elicit significant antibody titer. In contrast, the pdm09 vaccine alone elicited significant antibody titer in young adult mice. Antibody subtype analysis found that the pdm09 vaccine alone elicited Th2-biased antibody responses in young adult mice, while incorporation of the CpG 1018 adjuvant promoted the elicitation of potent Th1-biased antibody responses in aged mice. The pdm09 vaccine alone was further found to induce significant expansion of Th2 cells in young adult mice, while incorporation of the CpG 1018 adjuvant stimulated significant expansion of Th1 cells in aged mice. The CpG 1018 adjuvant also stimulated vaccine-specific cytotoxic T lymphocytes in aged mice. The pdm09 vaccine in the presence of CpG 1018 elicited significant protection against lethal viral challenges, while the pdm09 vaccine alone failed to confer significant protection in young adult or aged mice. Our study provided strong evidence to support the high effectiveness of the CpG 1018 adjuvant to boost influenza vaccination in aged mouse models.

Comparative tissue proteomics reveals unique action mechanisms of vaccine adjuvants

Yibo Li, Zhuofan Li, Xinyuan Chen

Radiofrequency adjuvant (RFA) was recently developed to boost influenza vaccination without the safety concerns of chemical adjuvants due to their physical nature. Yet, the action mechanisms of RFA remain largely unknown. Omics techniques offer new opportunities to identify molecular mechanisms of RFA. This study utilized comparative tissue proteomics to explore molecular mechanisms of the physical RFA. Comparison of RFA and chemical adjuvant (Alum, AddaVax, MPL, MPL/Alum)-induced tissue proteome changes identified 14 exclusively induced proteins by RFA, among which heat shock protein (HSP) 70 was selected for further analysis due to its known immune-modulating functions. RFA showed much weakened ability to boost ovalbumin and pandemic influenza vaccination in HSP70 knockout than wild-type mice, hinting crucial roles of HSP70 in RFA effects. This study supports comparative tissue proteomics to be an effective tool to study molecular mechanisms of vaccine adjuvants.

Impact of Advanced Glycation Endproduct Precursors on Erastin-Induced Ferroptosis in Human Keratinocytes: A Proteomic-based Ingenuity Pathways Analysis

Huifang Li, Ni Deng, Tess Puopolo, NavindraP. Seeram, Chang Liu, Hang Ma

Carbonyls are reactive compounds generated from the metabolism of carbohydrates, lipids, and amino acids, and can react with DNA to form glyoxal (GO) / methylglyoxal (MGO)-DNA adducts, causing oxidative DNA damage1. GO and MGO can also disrupt cell membranes, enzymes, and signaling pathways, leading to elevated oxidative stress and various forms of cell death, including ferroptosis2. Ferroptosis is a recently discovered iron-dependent form of cell death characterized by accumulated lipid peroxidation3. Herein, we aimed to depict the overall impact of GO and MGO on erastin-induced ferroptosis in human keratinocytes and investigated through a proteomics-based approach to identify differentially expressed proteins and pathways associated with pathological skin conditions.

Membrane vesicles as microbial delivery systems for mediating bacterial competition

Arvie Grace B. Masibag, Ololade Gbadebo, Marta Gomez-Chiarri, David R. Nelson, David C. Rowley

Membrane vesicles (MVs) are nanosized single- or double-membraned particles produced by bacteria through blebbing and, for some, explosive cell rupture. MVs may contain biologically active components, such as lipopolysaccharides (LPS), phospholipids, and outer membrane proteins (OMPs), as well as periplasmic components, allowing MVs to be involved in cell-to-cell communication, stress responses, antimicrobial resistance, horizontal gene transfer, and immune evasion. MVs can also package virulence factors and may function as delivery vesicles, contributing to host colonization and infection-associated pathology. MVs have been proposed as a vaccine platform technology to limit the severity of infectious disease outbreaks. In this study, MVs were isolated and purified from the marine bacteria Pseudoalteromonas piscicida JC3, Vibrio corallilyticus RE22, Vibrio parahaemolyticus PSU5579, and Phaeobacter inhibens S4. The MVs were subjected to an array of in vitro assays to assess their possible roles in microbial interactions. Spot plate antibacterial assays showed that P. piscicida JC3 possesses antibacterial potential against P. piscicida PSU5579 and Staphylococcus aureus. MVs from V. parahaemolyticus PSU5579 were found to reduce biofilm production by P. inhibens S4. Additionally, MVs from P. piscicida JC3 and P. inhibens S4 demonstrated potential for iron acquisition via siderophore production. These results demonstrate the production of MVs in phylogenetically diverse marine bacteria and indicate possible ecological roles for mediating microbial interactions and nutrient acquisition.

Effectiveness of Low-Dose Naltrexone as an Analgesic in the Treatment of Fibromyalgia: Systematic Review and Meta-analysis.

Abiodun Ologunowa, Marianne Otoo

Background: Fibromyalgia is a condition associated with pain, fatigue, sleep disturbance, and emotional and cognitive distress. Over 4 million individuals are affected by fibromyalgia in the US. Fibromyalgia is more prevalent in women compared to men(1). Naltrexone HCl is an opioid antagonist approved for treating alcohol and opioid use disorders. Low-dose naltrexone (LDN) is defined as Naltrexone HCl doses <10mg. LDN is currently used as an off-label medication in the management of fibromyalgia.

Method: This study was performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines in March -April 2023. Electronic databases (PubMed, Embase, Scopus, Cochrane Library) and gray literature (ProQuest, Clinicaltrial.gov) were searched using “Fibromyalgia” and “Low-dose naltrexone” search terms.

Results: 565 articles were identified from the database search. 8 articles (6 full papers and 2 abstracts) were identified for inclusion in the systematic review and meta-analyses (Five RCTs, and Three Observational Studies). In the analysis of pain score, there was a significant reduction in the post-LDN group relative to the baseline pain score by 0.72 [95%CI: -1.21, -0.22; I2= 64%]. Pain score was reduced by 0.31 in the LDN group compared to the placebo group [95%CI: -0.61, -0.01; I2=0%]. In the analysis of FIQ scores, the FIQ score was reduced by -1.08 [95%CI: -1.62, -0.55; I2=63%] post-LDN relative to the baseline. There was a non-significant reduction in FIQ score by 0.46 [-1.48, 0.56; I2=86%] in the LDN group compared to the placebo. In the subgroup analysis by full-text publications, the Mean Pain score and FIQ score were reduced by 0.59 [95%CI: -1.04, -0.14; I2=0%] and 0.82 [95% CI:-1.21, -0.42; I2=0%] respectively comparing the post-LDN to the pre-LDN.

Conclusion: Low-dose naltrexone is significantly associated with a reduction in pain and FIQ scores.

Uncovering the anti-inflammatory mechanisms of phenolic-enriched maple syrup extract in lipopolysaccharide-induced peritonitis in mice: Insights from data-independent acquisition proteomics analysis

Tess Puopolo, Huifang Li, Hang Ma, Joseph Schrader, Chang Liu, Navindra P. Seeram

Our group has previously reported on the phytochemical composition and biological activities of a phenolic-enriched maple syrup extract (MSX), which showed promising anti-inflammatory effects in several disease models including diabetes and Alzheimer’s disease. However, the efficacious doses of MSX and its molecular targets involved in the anti-inflammatory effects are not fully elucidated. Herein, the efficacy of MSX in a peritonitis mouse model was evaluated in a dose-finding study and the underlying mechanisms were explored using data-independent acquisition (DIA) proteomics assay. MSX (at 15, 30 and 60 mg/kg) alleviated lipopolysaccharide-induced peritonitis by reducing the levels of pro-inflammatory cytokines including interleukin-1 beta (IL-1β), IL-6, and tumor necrosis factor alpha (TNF-α) in the serum and major organs of the mice. Furthermore, DIA proteomics analyses identified a panel of proteins that were significantly altered (both up- and down-regulated) in the peritonitis group, which were counteracted by the MSX treatments. MSX treatment also modulated several inflammatory upstream regulators including interferon gamma and tumor necrosis factor. Ingenuity pathway analysis suggested that MSX may modulate several signaling pathways in the processes of initiation of cytokine storm, activation of liver regeneration, and suppression of hepatocyte apoptosis. Together, these proteomic and in-vivo findings indicate that MSX could regulate inflammation signaling pathways and modulate inflammatory markers and proteins, providing critical insight into its therapeutic potential.