Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Gschanes-Schweiger, P.
The role of neutrophil granulocytes on T cell function in
bronchial carcinoma (NSCLC)
Humanmedizin; [ Diplomarbeit ] Medizinische Universität Graz; 2022. pp. 86
[OPEN ACCESS]
FullText
- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Heinemann Akos
- Kargl Julia
- Altmetrics:
- Abstract:
- Introduction and Objectives: The presence of neutrophil granulocytes in the TME has been repeatedly described. Recent studies of patients with NSCLC have shown that tumor-associated neutrophils (TANs) show a negative correlation with T lymphocytes and have an immunosuppressive effect in which T lymphocytes are inhibited in their function to fight against tumor cells. The aim of this study was to complement existing data, which have verified the promotion of tumor growth by inhibition of CD8+ T cells through TANs, by performing co-culture experiments of human T cells together with neutrophils in vitro as part of an experimental setup. Materials and methods: Blood samples were collected from healthy donors and T cells and neutrophils were isolated for co-culture experiments. T lymphocyte-neutrophil co-cultures were measured after three and six days in a proliferation assay and after 24 hours in an apoptosis assay. Before seeding, naïve neutrophils were treated with supernatants of cell lines either from lung cancer (A549) or normal human bronchial epithelium (B2B) to polarize them towards TANs; for comparison, a negative control of mono-cultured T lymphocytes was performed. After culturing, T cells were stained for CD4 and CD8 to separate T cell subsets and with eF450 proliferation dye for proliferation experiments or with PI/Annexin V apoptosis dye for apoptosis experiments. T cell proliferation and apoptosis were analyzed using flow cytometry. For both assays, four samples under identical experimental conditions were used for statistical analysis. Results: The co-cultures differed from the negative control only to a small extent. Neither statistically significant differences in proliferation behavior nor in enhanced apoptosis counts in CD4+ or CD8+ T cells could be observed. Co-cultures with A549-treated neutrophils showed the biggest disruption of T lymphocytes in proliferation behavior and in the absolute percentage of apoptosis. Overall, the analysis delivered contradictory results. Discussion: These observations indicate that the methods are still too immature to substantiate a possible negative correlation between neutrophils and T cells in a human model. Furthermore, the p-value had suffered from low sample size. Particularly paradoxical results may point to necessary improvements in terms of single reagents, experimental setup or work steps.