Lipopolysaccharide-induced inflammatory modulation of the apoptotic pathways in human primary aniridia limbal stromal cells, in vitro.

Authors

Shanhe Liu, Shuailin Li, Shao-Lun Hsu, Fabian N Fries, Zhen Li, Swarnali Kundu, Berthold Seitz, Maryam Amini, Shweta Suiwal, Julia Zimmermann, Simon Trusen, Tanja Stachon, Nóra Szentmáry

Published in

PloS one. Volume 21. Issue 10. Pages e0361130. Epub Oct 27, 2026.

Abstract

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.
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.
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.
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.

PMID:
42945177
Bibliographic data and abstract were imported from PubMed on 28 Oct 2026.

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