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. 2025 Feb 5;26(3):1349.
doi: 10.3390/ijms26031349.

Histology of the Upper Gastrointestinal Tract, Morphometry and Lymphocyte Subpopulations of the Duodenal Mucosa: Insights from Healthy Individuals

Albert Martín-Cardona  1   2 , Anna Carrasco  1   2 , Carme Ferrer  3 , Clarisa González-Mínguez  3 , Luis Luizaga-Velasco  3 , Xavier Tarroch  3 , Gerardo Gonzalez-Puglia  1 , Eva Tristán  1   2 , Natalia Berenice Cardozo-Rembado  1 , Natàlia Pallarès  4 , Cristian Tebé  4 , Beatriz Arau  1   2 , Isabel Salvador  1 , Ingrid Fajardo  1 , Raimon Rifà  1 , Laura Ruiz  1   2 , Pablo Ruiz-Ramírez  1   2 , Sònia Fernández-Herrera  1 , Agnès Raga  1 , Montserrat Aceituno  1   2 , Yamile Zabana  1   2 , Carme Loras  1   2 , Mireia Fonolleda  5 , Jordi Roigé  6 , Fernando Fernández-Bañares  1   2 , Maria Esteve  1   2
Affiliations

Histology of the Upper Gastrointestinal Tract, Morphometry and Lymphocyte Subpopulations of the Duodenal Mucosa: Insights from Healthy Individuals

Albert Martín-Cardona et al. Int J Mol Sci. .

Abstract

The upper oesophagogastrointestinal (UEGI) tract histology, intestinal morphometry and lymphocyte subpopulations of healthy people is scarcely known. In research studies of inflammation involving the UEGI tract, there is a lack of adequate healthy controls. Aims: To evaluate the histology of the UEGI tract and the duodenal lymphocyte subpopulations of healthy volunteers and patients with gastroesophageal reflux disease (GERD), the latter to assess if it could replace healthy subjects. Healthy individuals were excluded if they had symptoms, comorbidities, pregnancy, toxics, medications or abnormal blood analysis. Subjects in both groups with abnormal duodenal intraepithelial lymphocyte (IEL) counts were also excluded. A total of 280 subjects were assessed, and 37 were included (23 healthy and 14 with GERD). The GERD group showed a higher IEL count (median [IQR]: 19.5 [17-22]), than healthy group: (15 [12-18]), p = 0.004. Eosinophils, mast cells and intestinal morphometry were similar in both groups. In the lamina propria, CD4+ T cells decreased (p = 0.008), and CD8+ T cells increased (p = 0.014). The total innate lymphoid cells (ILC) and CD3- cells decreased (p = 0.007) in GERD group compared to healthy controls. At the intraepithelial level, NKT cells increased (p = 0.036) and ILC3 decreased (p = 0.049) in the GERD group. This is the first study to comprehensively map the histology, morphometry and duodenal subpopulations of healthy volunteers to help define a "gold standard" of normality. The differences found between both groups suggest that, whenever possible, healthy subjects should be included in research studies. Alternatively, we can consider a well-defined homogenous group with GERD to serve as the control group.

Keywords: eosinophils; flow cytometry; healthy mucosa; innate lymphoid cells; intestinal morphometry; intraepithelial lymphocytes; mast cells; natural killer cells; γδ+ cells.

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Conflict of interest statement

A. Martín-Cardona has received financial support for conference attendance, educational activities and research support from AbbVie, Biogen, Faes Farma, Ferring, Jannsen, MSD, Pfizer, Takeda, Dr. Falk Pharma, Lilly and Tillotts. R. Rifà has served as a speaker for Advanz and has received financial support for conference attendance and educational activities from Gilead, AbbVie, Roche, Advanz and Kern Pharma. Y. Zabana has received support for conference attendance, speaker fees, research support and consulting fees from: AbbVie, Adacyte, Alfa-Sigma, Almirall, Amgen, Boehringer Ingelheim, Dr Falk Pharma, FAES Pharma, Fresenius Kabi, Ferring, Galapagos, Janssen, J&J, Kern, Lilly, MSD, Otsuka, Pfizer, Sanofi, Shire, Takeda, Tillots. M. Esteve has received financial support for conference attendance and research support from Abbvie, Biogen, Faes Farma, Ferring, Jannsen, MSD, Pfizer, Takeda and Tillotts. F. Fernández-Bañares has received financial support for conference attendance and research support from AbbVie, Biogen, Faes Farma, Ferring, Jannsen, MSD, Pfizer, Takeda and Tillotts. The remaining authors report no conflicts of interest.

Figures

Figure 1
Figure 1
Study flow chart. Abbreviations: GERD: gastroesophageal reflux disease; NSAIDs: nonsteroidal anti-inflammatory drugs.
Figure 2
Figure 2
Histological images of duodenal samples from healthy individuals. (A) Immunohistochemical staining of CD3+ lymphocytes in the duodenal mucosa (Marsh 0). (B) H&E staining showing sparse eosinophils in the duodenal mucosa (Marsh 0). (C) Immunohistochemical staining of C-kit showing sparse mast cells in the duodenal mucosa (Marsh 0). Abbreviations: H&E: haematoxylin and eosin.
Figure 3
Figure 3
Strip chart of villus height (μm) (A), crypt depth (μm) (B), villus height-to-crypt depth ratio (VCR) (C) and intraepithelial lymphocytes (D) in the villi of healthy individuals compared with those in the gastroesophageal reflux disease (GERD) group. The red dot indicates the median.
Figure 4
Figure 4
Haematoxylin and eosin staining of duodenal mucosa from healthy individuals (Marsh 0). The architecture of villi and crypts is preserved. The black arrows indicate two examples of villus height measurements (µm), and the orange arrows indicate two measurements of crypt depth (µm) to assess duodenal morphometry.
Figure 5
Figure 5
Main intestinal lymphocyte subpopulations in healthy individuals compared to those in the gastroesophageal reflux disease (GERD) group and the overlap between them.
Figure 6
Figure 6
Intestinal cytometry panel of major intestinal lymphocyte subpopulations in healthy individuals compared to the gastroesophageal reflux disease (GERD) group.
See this image and copyright information in PMC

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