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. 2025 Feb 20;15(3):329.
doi: 10.3390/life15030329.

Histopathologic Concerns and Diagnostic Challenges in Hirschsprung's Disease: An Eastern European Single-Center Observational Study

Emőke Horváth  1   2 , Zoltán Derzsi  3   4 , Eliza Löckli  5 , Gyopár-Beáta Molnár  2   6 , Zsolt Bara  3   4 , Evelyn Kovács  4 , Horea Gozar  3   4
Affiliations

Histopathologic Concerns and Diagnostic Challenges in Hirschsprung's Disease: An Eastern European Single-Center Observational Study

Emőke Horváth et al. Life (Basel). .

Abstract

Background: We proposed a comprehensive clinicopathological study involving the characterization of the study cohort and a comparative analysis of biopsies and surgical specimens from patients with Hirschsprung's disease. The study was complemented by the diagnostic value of calretinin, CD56, and S-100 immunohistochemistry.

Methods: Descriptive statistical analysis of diagnostic variables in the group of biopsy specimens (n = 32) and bowel resection specimens (n = 16) was performed. The pattern of calretinin and CD56 expression in Meissner's plexus elements was analyzed and the maximum thicknesses of the nerve fibers were measured using morphometry with S100-immunostained sections.

Conclusions: Coupled calretinin-CD56 immunohistochemistry is useful in diagnosing ganglion cell paucity biopsies or specimens with incomplete submucosa. In cases where there are no ganglion cells but there are calretinin-positive nerve fibrils in the lamina propria without nerve trunk (NT) hypertrophy, re-biopsy is the best solution. The significant differences in NT size between biopsies and surgical specimens highlight the importance of assessing NT diameter in all tissue samples examined.

Keywords: CD56 and S-100 immunohistochemistry; aganglionosis; calretinin; diagnostic algorithm in rectal biopsies; nerve trunk hypertrophy.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
(a–c) A histological section of a resected colon–aganglionic segment from a patient with HD. (a) Myenteric aganglionosis associated with nerve trunk hypertrophy (H&E staining; scale bar 100 µm). (b) The exact diameter of the nerve trunk was determined on S-100-immunostained sections using digital morphometry (×400 magnification). (c) Myenteric and submucosal nerve hypertrophy in aganglionosis (rectum defined as ≥40 µm in diameter) is considered to be supportive of the diagnosis of HD. Active mucosal inflammation (enterocolitis) may also be observed.
Figure 2
Figure 2
(a–d) Calretinin and CD56 immunohistochemistry in patients diagnosed with Hirschsprung’s disease. (a) A histological section of a resected colon: normal segment from a patient with HD showing calretinin/DAB-positive ganglion cells in the submucosa (red arrows) and their extended nerve fibers in the lamina propria (purple arrows). Scale bar: 50 µm. (b) CD56/DAB staining visualizes the ganglion cells of the muscularis propria and submucosa (yellow arrows). Scale bar: 100 µm. (b) CD56/DAB-positive ganglion cells of the muscularis propria and submucosa (yellow arrows). Scale bar: 100 µm. (c,d) Rectal biopsy of the patient showing disorganization of the muscularis mucosae and lymphocytic infiltrate in the mucosa (blue star), but with negative staining for calretinin (c) and CD56 immunohistochemistry (d). Scale bar: 500 µm. Note: m—mucosa; mm—muscularis mucosae; sm—submucosa.
Figure 3
Figure 3
Algorithm for the diagnosis of Hirschsprung’s disease in rectal biopsies.
See this image and copyright information in PMC

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