Lipopolysaccharide-induced inflammatory modulation of the apoptotic pathways in human primary aniridia limbal stromal cells, in vitro
- 1Dr. Rolf M. Schwiete Center for Limbal Stem Cell and Congenital Aniridia Research, Saarland University, Homburg/Saar, Germany
- 2Department of Experimental Ophthalmology, Saarland University, Homburg/Saar, Germany
- 3Department of Ophthalmology, Saarland University Medical Center, Homburg/Saar, Germany
Abstract
Purpose: This study investigated apoptosis-related responses to a standardized 24-hour lipopolysaccharide (LPS) challenge in primary limbal stromal cells (LSCs) derived from patients with congenital aniridia (AN-LSCs) and control donors. Methods: LSCs were isolated from limbal tissue obtained from patients with congenital aniridia and control donors. Cells were treated with 0, 2.5, 10, and 17.5 µg/mL LPS for 24 hours. Apoptosis was assessed using Annexin V-APC/PI staining and flow cytometry. Apoptosis-related mRNA levels were quantified by qPCR, cell-associated protein fluorescence was assessed by flow cytometry, and secreted TNF-α was measured by ELISA. Results: The mean basal apoptosis rate was numerically higher in AN-LSCs than in control LSCs (2.10% vs. 0.52%), although the donor-level difference was not statistically significant. The exploratory linear mixed-effects model showed a significant main effect of LPS condition and a significant Group × LPS interaction. Apoptosis increased significantly at 10 and 17.5 µg/mL LPS compared with the corresponding untreated condition in both groups, and the increase from the untreated condition to 17.5 µg/mL LPS was significantly greater in AN-LSCs than in control LSCs (adjusted P = 0.003). qPCR and flow-cytometry analyses identified marker-specific LPS-associated changes, but no molecular Group × LPS interaction reached statistical significance. Cell-associated TNF-α fluorescence was distinguished from secreted TNF-α measured by ELISA. Conclusions: Acute LPS exposure increased apoptosis in both groups, with a greater increase in AN-LSCs at the highest concentration. Molecular changes were marker- and concentration-specific without evidence of differential molecular response patterns between groups. Given the small number of independent donors, the marked donor-age imbalance, and the acute nature of the model, these findings should be considered exploratory and hypothesis-generating and require confirmation in a larger independent cohort of appropriately age-matched donors.
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BibTeX
@article{liu2026lipopolysaccharideinduced,
title = {{Lipopolysaccharide-induced inflammatory modulation of the apoptotic pathways in human primary aniridia limbal stromal cells, in vitro}},
author = {Shanhe Liu and Shuailin Li and Shao-Lun Hsu and Fabian N. Fries and Zhen Li and Swarnali Kundu and Berthold Seitz and Maryam Amini and Shweta Suiwal and Julia Zimmermann and Simon Trusen and Tanja Stachon and Nóra Szentmáry},
journal = {PLOS One},
year = {2026},
month = oct,
doi = {10.1371/journal.pone.0361130},
url = {https://doi.org/10.1371/journal.pone.0361130},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Lipopolysaccharide-induced inflammatory modulation of the apoptotic pathways in human primary aniridia limbal stromal cells, in vitro
AU - Shanhe Liu
AU - Shuailin Li
AU - Shao-Lun Hsu
AU - Fabian N. Fries
AU - Zhen Li
AU - Swarnali Kundu
AU - Berthold Seitz
AU - Maryam Amini
AU - Shweta Suiwal
AU - Julia Zimmermann
AU - Simon Trusen
AU - Tanja Stachon
AU - Nóra Szentmáry
PY - 2026
DA - 2026/10/27
JO - PLOS One
DO - 10.1371/journal.pone.0361130
UR - https://doi.org/10.1371/journal.pone.0361130
AB - Purpose: This study investigated apoptosis-related responses to a standardized 24-hour lipopolysaccharide (LPS) challenge in primary limbal stromal cells (LSCs) derived from patients with congenital aniridia (AN-LSCs) and control donors. Methods: LSCs were isolated from limbal tissue obtained from patients with congenital aniridia and control donors. Cells were treated with 0, 2.5, 10, and 17.5 µg/mL LPS for 24 hours. Apoptosis was assessed using Annexin V-APC/PI staining and flow cytometry. Apoptosis-related mRNA levels were quantified by qPCR, cell-associated protein fluorescence was assessed by flow cytometry, and secreted TNF-α was measured by ELISA. Results: The mean basal apoptosis rate was numerically higher in AN-LSCs than in control LSCs (2.10% vs. 0.52%), although the donor-level difference was not statistically significant. The exploratory linear mixed-effects model showed a significant main effect of LPS condition and a significant Group × LPS interaction. Apoptosis increased significantly at 10 and 17.5 µg/mL LPS compared with the corresponding untreated condition in both groups, and the increase from the untreated condition to 17.5 µg/mL LPS was significantly greater in AN-LSCs than in control LSCs (adjusted P = 0.003). qPCR and flow-cytometry analyses identified marker-specific LPS-associated changes, but no molecular Group × LPS interaction reached statistical significance. Cell-associated TNF-α fluorescence was distinguished from secreted TNF-α measured by ELISA. Conclusions: Acute LPS exposure increased apoptosis in both groups, with a greater increase in AN-LSCs at the highest concentration. Molecular changes were marker- and concentration-specific without evidence of differential molecular response patterns between groups. Given the small number of independent donors, the marked donor-age imbalance, and the acute nature of the model, these findings should be considered exploratory and hypothesis-generating and require confirmation in a larger independent cohort of appropriately age-matched donors.
SN - 1932-6203
ER -