LPS induced inflammatory responses in human peripheral blood mononuclear cells is mediated through NOX4 and Giα dependent PI-3kinase signalling
Open accessAbstract
COPD is a disease of innate immunity and bacterial infections are a dominant cause of exacerbations in the later stages resulting in poor health and high mortality. The pathogen-associated molecular pattern (PAMP) lipopolysaccharide (LPS) is sensed by immune cells through activation of the toll-like receptor 4 (TLR4). This leads to the activation of NADPH oxidase (NOX) and NF-κB which together drive COPD inflammation. In this study we show in human PBMCs that LPS stimulated proinflammatory cytokine release (CXCL8 and IL6) was inhibited by approximately 50% by the broad specificity phosphatidylinositol 3-kinase (PI3K) inhibitor, wortmannin. Our results also demonstrate that activation of PI3K following LPS stimulation is mediated by a NOX4 dependent mechanism releasing endogenous H 2 O 2, as the NOX4 inhibitor apocynin blocked LPS induced AKT phosphorylation. Moreover, LPS-induced PI3K activation was inhibited by the anti-oxidant N-acetylcysteine in a concentration dependent manner (IC 50~100 μM). In addition, our data demonstrated that inhibition of small G proteins, by pre-treatment with pertussis toxin, inhibited LPS-induced AKT phosphorylation. Furthermore, the G-protein inhibitors pertussis toxin and mastoparan both inhibited LPS-induced CXCL8 and IL-6 release by approximately 50%. Together, these data indicate there is a mechanism in human PBMCs where TLR4 activation by LPS leads to ROS generation through NOX4 and activation of the PI3K pathway. This effect is apparently mediated through small G proteins facilitating the release of pro-inflammatory cytokines.
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Treatment of chimeric mice with LPS led to a similar increase in the number of Trp53-mutant PB myeloid cells and LSKs.
“Treatment of chimeric mice with LPS (Fig. 5a), which induces an inflammatory response mediated through the release of IL1β and IL6 (ref. 37), among others, led to a similar increase in the number of Trp53-mutant PB myeloid cells and LSKs (Fig. 5f,g).”
From Single-cell multi-omics identifies chronic inflammation as a driver of TP53-mutant leukemic evolution (2023) · cites this paper as [37]
The citation is attached to the explanation that LPS induces inflammation through IL-1β and IL-6 release, not to the citing authors’ finding about Trp53-mutant cells. The Results and Discussion report LPS-induced IL-6 release from human PBMCs, but not IL-1β release; the Introduction says only that LPS mimics some effects of IL-1β. No page numbers are available in our copy. (LLM, conf=0.88)
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The phosphoinositide 3-kinase pathway
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Rodriguez-Meira, A., Norfo, R., Wen, S., Chédeville, A. L., Rahman, H., O’Sullivan, J., Wang, G., Louka, E., Kretzschmar, W. W., Paterson, A., et al. (2023) · Nature Genetics · doi:10.1038/s41588-023-01480-1 · Cited by 158 · see citing works
Cataldi, S., Bonavolontà, V., Poli, L., Clemente, F. M., De Candia, M., Carvutto, R., Silva, A. F., Badicu, G., Greco, G., & Fischetti, F. (2022) · Biology · doi:10.3390/biology11030479 · Cited by 68 · see citing works
- Authors
- Ngkelo, A., Meja, K., Yeadon, M., Adcock, I., & Kirkham, P. A.
- Affiliations
- Imperial College London (GB)
- University College London (GB)
- Pfizer (United Kingdom) (GB)
- Corresponding author
- Kirkham, Paul A
- Venue
- Journal of Inflammation
- Published
- 2012-01-12
- Volume
- 9
- Issue
- 1
- Pages
- 1-1
- ISSN
- 1476-9255 (ISSN-L)
- PMC
- PMC3293082
- OpenAlex
- W2125496800
- Cited by
- 238
- References
- 1
- Citations by year
- 2026: 7 · 2025: 19 · 2024: 29 · 2023: 26 · 2022: 21 · 2021: 30 · 2020: 20 · 2019: 9 · 2018: 10 · 2017: 11 · 2016: 16 · 2015: 15 · 2014: 13 · 2013: 7 · 2012: 4
- FWCI
- 5.46
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Journal of Inflammation (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Spiral (Imperial College London) (submittedVersion, other-oa)
- Funders
- Pfizer
- Wellcome Trust
- National Institute for Health and Care Research (NF-SI-0508-10212)
- Medical Research Council
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
- Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- lipopolysaccharide, NOX4, PI3K signaling, small GTPases, pertussis toxin, mastoparan, Akt phosphorylation, reactive oxygen species, hydrogen peroxide, TLR4, chronic obstructive pulmonary disease, peripheral blood mononuclear cells, interleukin-8, interleukin-6, inflammatory cytokines, wortmannin, apocynin, N-acetylcysteine, innate immunity
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