Local Mast Cells and Neutrophils in Colorectal Cancer | Areti Barmpa
Distribution and prognostic value of tumor-infiltrating mast cells and neutrophils in colorectal cancer patients with liver metastases
Authors: Areti Barmpa, Denis Kucharik, Wenjing Ye (1), Filip Ambrożkiewicz (1), Václav Liška (1, 2),
Kari Hemminki (1)
Supervisor: Andriy Trailin (1)
(1) Biomedical Center, Faculty of Medicine in Pilsen, Charles University (2) Department of Surgery,
Faculty of Medicine in Pilsen, Charles University and University Hospital, Pilsen
State-of-the-Art: Colorectal cancer (CRC) is among the most common and lethal cancers globally.
The presence of liver metastasis (LM) is associated with a significantly poorer prognosis, exposing the limitations of current prognostic tools. While immune profiling of T-cells in tumor microenvironment has improved prognostication, the role of innate immune cells in CRC and adjacent normal mucosa (NM) remains underexplored. Both mast cells and neutrophils play crucial roles in the NM of the colon, where they contribute to immune surveillance and tissue homeostasis. In CRC, both mast cells and neutrophils have been shown to play dual roles, either promoting or inhibiting tumor progression.
Objective: We aimed to quantitatively assess the distribution of mast cells and neutrophils in
the NM and primary tumor of CRC patients with synchronous LM, and to establish prognostic
associations of immune cell densities.
Material and Methods: We enrolled into this retrospective cohort study 55 patients, who underwent
resection of both primary CRC (pCRC) and synchronous LM. Formalin-fixed paraffin-embedded tissue sections from NM and pCRC were stained by immunoperoxidase method for mast cells with anti-CD117 antibodies and for neutrophils using anti-CD66b antibodies. Densities of immune cells per mm2 of the tissue were assessed in NM and tumor center (TC), inner margin (IM), outer margin (OM) and peritumor region (PT) of pCRC by using the QuPath image analysis software. Then cell densities were tested as prognostic variables for disease-free survival (DFS) by Cox regression analysis. A two-sided p value <0.05 was considered statistically significant.
Results & Discussion: Mast cell density was significantly greater in NM (median: 212 cells/mm²)
compared to TC (36 cells/mm²), as well as in OM (76 cells/mm²) and PT (85 cells/mm²) of pCRC vs
TC and IM (30 cells/mm²). Neutrophil density was significantly greater in the TC (56 cells/mm²)
compared to NM (12 cells/mm²). Within pCRC neutrophil density did not differ significantly between
TC, IM (58 cells/mm²), OM (42 cells/mm²) and PT (36 cells/mm²). We observed significant correlation between neutrophils and mast cells in OM (Spearman Rho=0.32) and PT (Rho=0.30) of pCRC. High densities of neutrophils in PT of pCRC were associated with longer DFS (Hazard ratio 0.44, 95% confidence interval 0.22–0.88, p=0.021), but we didn’t find associations between mast cell densities in any region with outcomes.
Conclusion: Our study found differential distribution of innate immune cells between NM and
CRC with mast cells predominating in NM and neutrophils in TC of pCRC. Neutrophil infiltration
in the PT region of pCRC in patients with synchronous LM confers a survival benefit, warranting
further investigation.
Funding: AZV NW24-03-00521
Study program: Master´s degree – General Medicine | Year of study: 4
ID: 1110
Dendritic Cells in Colorectal Cancer | Nishant Kumar
Distribution of immature and mature dendritic cells in colorectal cancer patients with liver metastases
Authors: Nishant Kumar (1), Jehnnia Mikaelly Liver Freitas (1), Esraa Ali (1), Sergii Pavlov (1), Aylin Sare Naz Uysal (1), Filip Ambrożkiewicz (1), Václav Liška (1,2), Dmytro Robota (3), Kari Hemminki (1)
Supervisor: Andriy Trailin (1)
(1) Biomedical Center, Faculty of Medicine in Pilsen, Charles University (2) Department of Surgery, Faculty of Medicine in Pilsen, Charles University and University Hospital, Pilsen (3) Zaporizhzhia State Medical and Pharmaceutical University
State-of-the-Art: Colorectal cancer (CRC) is a leading cause of cancer-related deaths in Europe, with metastatic disease being the main driver of mortality. The liver is the most common site of metastases from CRC. T-cell infiltration in primary CRC significantly impacts clinical outcomes. Owing to their ability to orchestrate tolerogenic or immunogenic responses to tumor antigens, antigen-presenting dendritic cells (DCs) play a pivotal role in anti-cancer immunity. Multifaceted functions of DCs are acquired through a complex, multistage process called maturation. Immature DCs facilitate tolerance, while mature DCs strongly promote anti-tumor immunity. With impact on infiltration of other leukocytes, tumor associated DCs have been correlated with patient prognosis in CRC.
Objective: We aimed to explore the distribution of immature and mature DCs in primary CRC along with adjacent normal mucosa (NM) and to investigate the prognostic value of DCs in CRC patients with synchronous liver metastases.
Material and Methods: Paired formalin-fixed paraffin-embedded specimens of primary CRC and tumor-adjacent NM were collected from patients who underwent resection of both primary tumor and synchronous liver metastases (n = 55). Immunohistochemical staining followed by computer-assisted image analysis were performed to quantify CD1a+ immature and CD208+ mature DCs in the NM, tumor center, inner margin, outer margin, and peritumor regions of the primary CRC. DC densities across these regions were investigated and tested as prognostic variables for time to recurrence, disease-free survival and overall survival.
Results & Discussion: CD1a+ DCs displayed greater densities in TC compared to the NM (median 6 vs. 0 cells/mm², P<0.001), whereas density of CD208+ DCs was non-significantly greater in NM (13 vs 8 cells/mm², P=0.06). For CD1a+ DCs, the IM exhibited greater densities than the TC and OM (10 vs. 6 and 5 cells/mm² respectively, P<0.05), whereas the PT harbored the lowest densities (1 cell/mm²). As for CD208+ DCs, the OM displayed significantly greater densities (21 cells/mm²) compared to other regions of pCRC (TC: 8, IM: 6, PT: 13 cells/mm², P<0.05). A higher infiltration of CD1a+ DCs significantly correlated with greater densities of CD208+ DCs in the TC, IM, and OM, highlighting an interplay between these cell types. However, no significant associations were found between survival outcomes and either type of DC
Conclusion: Our study has profiled DCs in normal mucosa and primary CRC tissue showing significant heterogeneity for both immature and mature cell types. Assessing the prognostic value of DCS requires their detailed profiling in liver metastases.
Funding: AZV NW24-03-00521.
Study program: Master´s degree – General Medicine | Year of study: 4
ID: 1125
Lymphocytes and Endometriosis | Žaneta Míšková
Lymphocyte Subpopulations in Women with Endometriosis
Authors: Žaneta Míšková (1), Zuzana Prášilová (1), Barbora Sedláčková (1), Lukáš Prajka (2), Monika
Holubová (3,4)
Supervisor: Miroslava Čedíková (1,3)
(1) Department of Physiology, Faculty of Medicine in Pilsen, Charles University (2) Department
of Gynaecology and Obstetrics, Faculty of Medicine in Pilsen, Charles University and University
Hospital, Pilsen (3) Laboratory of Tumor Biology and Immunotherapy, Biomedical Center, Faculty
of Medicine in Pilsen (4) Department of Haematology and Oncology, Faculty of Medicine in
Pilsen, Charles University and University Hospital, Pilsen
State-of-the-Art: Endometriosis is a chronic, oestrogen-dependent, inflammatory and systemic
disease characterized by the presence of endometrial glands and stroma outside the uterine cavity. It affects approximately 10–15% of women of reproductive age and significantly reduces quality of life due to pain and infertility. Despite its benign nature, the exact cause of endometriosis remains unclear. Increasing evidence points to an important role of the immune system in its pathogenesis, with abnormalities observed in both local and systemic immune responses. However, the precise mechanisms of immune involvement in disease development are still not fully understood.
Objective: This study aimed to analyse lymphocyte subpopulations in the peripheral blood of
women with ovarian endometriosis vs. healthy controls, focusing on T cell distribution/activation
and inhibitory KIR receptor expression on NK cells to reveal immune differences.
Material and Methods: Peripheral blood samples were obtained from 24 women with confirmed
ovarian endometriosis and 29 healthy female controls. All participants completed a standardized questionnaire collecting clinical and gynaecological history, including age, weight, height, body mass index, age at menarche, menstrual cycle length, and duration of menstrual bleeding. Peripheral blood mononuclear cells (PBMCs) were isolated and analysed by flow cytometry to determine the representation and activation status of T lymphocyte subpopulations (naive, memory, cytotoxic CD8⁺, helper CD4⁺, double-negative CD4⁻CD8⁻, and regulatory T cells) and the expression of selected inhibitory KIR receptors (2DL1, 2DL2, 2DL3, 2DL5, 3DL1, 3DL2) on NK cells.
Results & Discussion: No statistically significant differences were found in demographic or gynaecological
characteristics between the two groups. The distribution of T lymphocyte subpopulations did not significantly differ; however, the control group showed a higher proportion of activated T cells, suggesting possible functional impairment in the patient group. In contrast, the expression of the inhibitory receptor KIR2DL2 on NK cells was significantly increased in patients with endometriosis, which may contribute to reduced NK cell cytotoxic activity and facilitate the implantation and survival of ectopic endometrial tissue.
Conclusion: These findings suggest that immune alterations in endometriosis may be more
related to impaired cell functionality than to absolute cell counts. Reduced T cell activation and
increased inhibitory NK receptors may enable persistence of ectopic endometrial tissue.
Funding: This study was supported by the grant of the Ministry of Health of the Czech Republic –
Conceptual Development of Research Organization (Faculty Hospital in Pilsen – FNPl, 00669806).
Study program: Master´s degree – General Medicine | Year of study: 3
ID: 1134
A Decade of Imported Infectious Diseases | Viktorie Goncharuk
Imported Infectious Diseases in a Faculty Hospital: A 10-Year Retrospective Study
Author: Viktorie Goncharuk (1)
Supervisor: Dalibor Sedláček (1)
(1) Department of Infectious Diseases and Travel Medicine, Faculty of Medicine in Pilsen, Charles University and University Hospital, Pilsen
State-of-the-Art: Imported infectious diseases are a significant part of healthcare systems, especially with increasing global travel. While the COVID-19 pandemic temporarily reduced travel and imported infections, case numbers have risen again post-pandemic. This study analyzes data from a faculty hospital over the past 10 years highlighting notable cases to enhance awareness and preparedness in clinical practice.
Objective: To analyze the occurrence of imported infectious diseases treated at a faculty hospital over a 10-year period (2015–2024) and highlight clinically relevant cases that may serve as educational examples for improving diagnostic vigilance.
Material and Methods: This retrospective study utilized data extracted from the hospital‘s electronic medical records from January 2015 to December 2024. Diagnoses were identified based on codes associated with imported infectious diseases. For each case, the following parameters were recorded: year, diagnosis, outpatient vs. inpatient care and frequency. Statistics (absolute frequencies, trend visualization in line charts) were used to analyze temporal changes. Selected cases of malaria, giardiasis, dengue, and hepatitis E were further described due to their clinical significance.
Results & Discussion: The analysis of the data revealed a noticeable increase in imported infectious diseases in post-COVID era, particularly cases of giardiasis, malaria, dengue, hepatitis E, and enterotoxigenic E. coli infections. Diagnosing imported infectious diseases can be challenging due to their diverse presentations, but it’s important for clinicians to be aware of the most common ones seen in our hospital to ensure timely and accurate diagnosis.
Conclusion: The sharp rise in dengue and malaria cases in the last three years corresponds with global post-pandemic travel recovery. Hepatitis E remains the most common imported infection, likely linked to food-related exposure during travel. Our hospital’s data aligns with trends seen in other European center.
Study program: Master´s degree – General Medicine | Year of study: 3
ID: 1113