Teipel, Flavio Jan; Teipel, Flavio: Influence of a dietary supplement with conjugated linoleic acid (CLA) on systemic immune responses in patients with multiple sclerosis. 2020
Inhalt
- Abbreviations
- 1 Introduction
- 1.1 Multiple sclerosis
- 1.2 Conjugated linoleic acid
- 1.2.1 Chemical aspects
- 1.2.2 Occurrence in food items and natural intake
- 1.2.3 Effects in the context of inflammatory diseases
- 1.2.3.1 Effects on the immune system
- 1.2.3.2 Effects on inflammatory diseases
- 1.2.3.3 Effects in the context of CNS autoimmunity
- 1.2.4 Discussed modes of action
- 1.3 Aim of the thesis
- 2 Material
- 2.1 Devices
- 2.2 Disposables
- 2.3 Study medication
- 2.4 Constituents of media and buffers
- 2.5 Composition of media and buffers
- 2.6 Chemicals and reagents
- 2.7 Assay kits
- 2.8 Antibodies for cell activation
- 2.9 Antibodies for flow cytometry
- 2.10 Software
- 3 Methods
- 3.1 Biological sample acquisition
- 3.1.1 Clinical study
- 3.1.1.1 Inclusion and exclusion criteria
- 3.1.1.2 Study design
- 3.1.1.3 Analysis of baseline demographical and clinical data
- 3.1.1.4 Analysis of natural fat and CLA intake
- 3.1.2 HC samples
- 3.2 Cell culture
- 3.2.1 PBMC isolation
- 3.2.2 PBMC thawing
- 3.2.3 Determination of cell concentrations
- 3.2.4 Adjustment of cell concentrations
- 3.2.5 CD4+ T cell isolation
- 3.2.6 Cell activation
- 3.2.6.1 PHA activation
- 3.2.6.2 LAC activation
- 3.2.6.3 PMA / ionomycin activation
- 3.2.6.4 anti-CD3 and anti-CD28 activation
- 3.2.7 in vitro CLA Treatment
- 3.3 Analytical methods
- 3.3.1 Enzyme-linked immunosorbent assay (ELISA)
- 3.3.2 Luminex® assay
- 3.3.3 Propidium iodide (PI) staining
- 3.3.4 Flow cytometry
- 3.3.4.1 Surface staining at 4 C
- 3.3.4.2 Surface staining at 37 C
- 3.3.4.3 Combined surface and intracellular staining at 4 C
- 3.3.4.4 Combined surface and intracellular staining at 37 C / 4 C
- 3.3.4.5 Combined surface and intracellular staining at 4 C after LAC stimulation
- 3.3.4.6 Data analysis
- 3.3.5 Metabolism analysis
- 3.4 Statistical methods
- 4 Results
- 4.1 in vitro experiments
- 4.1.1 Cytotoxicity investigation
- 4.1.2 CD4+ T cell cytokine production upon activation
- 4.1.3 CD4+ T cell metabolism
- 4.2 Clinical study
- 4.2.1 Baseline demographical and clinical data
- 4.2.2 Natural fat and CLA intake
- 4.2.3 Clinical study endpoints
- 4.2.3.1 Adherence to study medication
- 4.2.3.2 Relapses
- 4.2.3.3 EDSS during follow-up
- 4.2.3.4 Paraclinical data
- 4.2.3.5 Correlation of relapses, EDSS during follow-up and paraclinical data
- 4.2.3.6 Adverse events
- 4.2.4 In-depth functional immune phenotyping
- 4.2.5 CD4+ T cell cytokine production upon activation
- 4.2.6 CD4+ T cell metabolism
- 5 Discussion
- 5.1 Clinical data
- 5.1.1 Baseline demographical and clinical data
- 5.1.2 Natural fat and CLA intake
- 5.1.3 Adherence to study medication
- 5.1.4 Relapses, EDSS and paraclinical development during follow-up
- 5.1.5 Adverse events
- 5.2 In-depth functional immune phenotyping
- 5.2.1 CD4+ T cells
- 5.2.2 CD8+ T cells
- 5.2.3 B cells
- 5.2.4 CD4+ Treg cells
- 5.2.5 Methodological limitations
- 5.3 CD4+ T cell cytokine production upon activation
- 5.4 CD4+ T cell metabolism
- 5.5 Study limitations
- 5.6 Gut microbiota
- 5.7 Outlook
- 6 Summary
- Figures
- Tables
- Literature
- Acknowledgments
- Curriculum vitae
- Appendix