Quantitative pathology characterization of T cells, B cells, and neutrophils in gastric biopsies with Helicobacter pylori infection

Helicobacter pylori (HP) infection is a major driver of chronic gastritis and is strongly associated with the development of preneoplastic lesions and gastric cancer. The host immune response, including both activation and regulatory mechanisms, plays a critical role in disease progression and tissue damage. To characterize the quantitative profile of T lymphocytes, B lymphocytes, and neutrophils in gastric biopsies with and without HP infection using digital quantitative pathology. A cross-sectional analytical observational study included 154 patients with gastric biopsy specimens classified as HP -positive or negative. Dual immunohistochemistry was performed for CD4/CD8, CD20, and MPO, with HP immunohistochemistry used as part of infection assessment. Whole-slide images were digitized and analyzed in QuPath for automated cell quantification. A multivariable logistic regression model was applied with HP status as the dependent variable. A total of 154 patients were included, of whom 76 were HP-positive. In multivariable analysis, activity (neutrophilic infiltration) (OR: 28.5; 95% CI: 11.1–73.5; p < 0.001) and the presence of lymphoid follicles (OR: 3.43; 95% CI: 1.33–8.88; p = 0.011) were significantly associated with infection. HP-positive cases showed a higher proportion of diffuse CD20 + B cells in the restricted non-aggregate analysis (64.54% vs. 29.79%, p < 0.001). The CD4:CD8 ratio was numerically higher in HP-positive than in HP-negative patients (median, 3.73 vs. 3.16), although this difference was not statistically significant ( p = 0.756); similarly, CD4 + and CD8 + cell percentages did not differ significantly between groups. HP infection was associated with increased neutrophilic infiltration and B-cell-associated changes. Although the CD4:CD8 ratio was numerically higher in HP-positive patients, the difference was not statistically significant, and CD4 + and CD8 + cell percentages were also comparable between groups. Quantitative digital pathology provides an objective approach for characterizing inflammatory-cell populations in the gastric mucosa and may complement conventional histopathological assessment.