Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Kriso, V.
Characterisation of the lymphocytic infiltrate in skin biopsies of scleroderma patients before and during longtime B-cell-depletion treatment
Humanmedizin; [ Diplomarbeit ] Graz Medical University; 2018. pp. 73
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Moazedi-Fürst Florentine
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- Abstract:
- Objective: The aim of this thesis was to establish a staining technique in order to evaluate the amount of Th17 cells in the skin of patients suffering from Systemic Sclerosis before and during long time CD20 – depletion therapy with Rituximab and compare the results to healthy controls. Introduction: Systemic Sclerosis is a rare, however, devastating connective tissue disease, which untreated, can lead to an involvement of the inner organs (lung, heart) and death. As the immune system plays an important part in the pathogenesis of the disease by overreacting and stimulating excessive fibrosis, immunomodulatory therapeutics such as Rituximab are being used and show promising improvement of the clinical state of the patients. It is discussed in the literature that also the Th17 cells of the immune system contribute to the disease and therefore we decided to take a closer (microscopical) look at the skin lesions before and during the treatment with Rituximab regarding the presence of these cells. Material and Methods: The technique to perform a double immunohistochemistry on paraffin embedded skin sections for Th17 cells was established. Patients, having already provided a skin sample before the initiation of the CD20 – depletion treatment in the study (EK. number 24-184 ex 11/12), were asked to give another sample at the earliest one year after the first biopsy (EK-No.: 30-091 ex 17/18). The specimens were further processed in the laboratory of the department of Rheumatology and immunohistochemically double and single stained. Possible influencing factors (type of Systemic Sclerosis, additional immunosuppressive therapy) were assessed. The single stained specimens are going to be scanned and overlaid on the computer to find the double stained Th17 cells. Results: 7 patients participated in this pilot study. Due to the overlay of the chromogens on the surface (CD4) and cytoplasmic (IL-17) working antibodies, it was not possible to reliably and objectively interpret the results of the double staining immunohistochemistry. The single staining of the CD4+ cells showed that the amount of the T helper cells is increased in SSc patients compared to healthy controls. Regarding a statement concerning the Th17 subgroup, the computer scans need to be awaited and interpreted. However, as an additional finding we noticed IL7+ mast cells. Discussion: Despite great challenges, the immunohistochemical staining method was established. We found IL-17 expressing mast cells, which proves that the single staining worked and it is only a matter of time until interesting results regarding the Th17 cells will follow. However, the delay in schedule prevented possible findings from being presented in this thesis. They will be discussed in a separate scientific work. Additionally, it should be mentioned that it might be of interest to investigate the stained mast cells regarding their development during RTX-treatment in combination with the IL-17-levels.