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. 2025 Apr 28:18:1562954.
doi: 10.3389/fnmol.2025.1562954. eCollection 2025.

Traumatic brain injury and autophagy: a pilot study about the immunohistochemical expression of LC3B, Beclin 1, p62, and LAMP2A in human autoptic samples

Tommaso Livieri  1 , Letizia Alfieri  2 , Emiliana Giacomello  1 , Djordje Alempijević  3 , Tijana Petrovic  3 , Yanko Georgiev Kolev  4 , Davide Radaelli  1 , Margherita Neri  2 , Stefano D'Errico  1
Affiliations

Traumatic brain injury and autophagy: a pilot study about the immunohistochemical expression of LC3B, Beclin 1, p62, and LAMP2A in human autoptic samples

Tommaso Livieri et al. Front Mol Neurosci. .

Abstract

Introduction: Autophagy is a cellular stress response that has been shown in the literature to be active in cerebral cells after a traumatic brain injury (TBI). The aim of this study is to investigate the potential use of four proteins involved in autophagy (LC3B, Beclin 1, p62, and LAMP2A), as a forensic diagnostic marker for TBI.

Methods: We analyzed histological samples obtained from the frontal lobe of 10 subjects who died within 1 h of a TBI (Group A), 13 who died between 1 h and 32 days post-TBI (Group B), and a control group of 10 subjects who died without head trauma (Group C). Immunohistochemical (IHC) staining using anti-LC3B, anti-Beclin 1, anti-p62 and anti-LAMP2A antibodies was performed.

Results and discussion: The results show that LC3B staining was the only one that show a statistically significant difference between groups. In particular, the percentage of neurons displaying an autophagic pattern was calculated from six random acquisitions per subject, and the results were compared across groups using one way ANOVA. Significant differences were observed between Groups A and B, and between Groups B and C, with p-values of 0.0055 and 0.0035, respectively. While the difference between Groups A and C was not statistically significant (p-value of 0.9845). These findings suggest that LC3B may serve as a useful diagnostic marker for TBI in cases where death is not immediate and open the door for further research.

Keywords: Beclin 1; LAMP2A; LC3B; autophagy; forensic; immunohistochemistry; p62; traumatic brain injury.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.

Figures

Figure 1
Figure 1
Three brain sections stained with anti-LC3B antibody. (A) Section from a case of Group A showing neurons with diffuse cytoplasmic staining (green arrows). (B) Section from a case of Group B, displaying neurons with staining indicative of an ongoing autophagic process (black arrows), characterized by vesicular cytoplasmic staining and perinuclear reinforcement (blue arrows). (C) Section from a control sample (group C), showing neurons with diffuse cytoplasmic staining (red arrows). (D) Distribution of mean percentage of neurons exhibiting an autophagic pattern in LC3B staining for each group (**p < 0.01).
Figure 2
Figure 2
Three brain sections stained with anti-p62 antibody. (A) Section from a case of Group A showing neurons with diffuse cytoplasmic staining (green arrows). (B) Section from a case of Group B, displaying neurons with staining potentially indicative of an ongoing autophagic process (black arrows), characterized by vesicular pattern localized to a single pole of the cytoplasm (blue arrows). (C) Section from a control sample (group C), showing neurons with diffuse cytoplasmic staining (red arrows). (D) Distribution of mean percentage of neurons exhibiting an autophagic pattern in p62 staining for each group (**p < 0.01).
See this image and copyright information in PMC

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