Skip to main content

Table 2 Endpoints of the study

From: Signatures of immune reprogramming in anti-CD52 therapy of MS: markers for risk stratification and treatment response

Primary Endpoints
• T cell subsets:
- CD4 and CD8 positive T cells: naïve T cells, T effector cells, T memory cells, regulatory T cells
- T-helper subsets: Th1, Th2, Th17
• B cell subsets:
- Recent bone marrow emigrants, mature naïve, memory B cells
- Plasma cells
• Natural killer cells:
- CD56bright, CD56dim
- Natural killer T cells
• Antigen-presenting cells:
- Dendritic cells: CD303+ plasmacytoid, CD11c+ and CD141+ myeloid dendritic cells
- Monocytes and macrophages
• Myeloid-derived suppressor cells
• Markers of autoimmunity in the serum (every 6 months):
- ANA, cANCA, pANCA, anti-dsDNA, anti-TSHR, anti-TPO, RF, anti-CCP, anti-GBM, anti-platelet antibodies
Secondary Endpoints
• Activation status of cell surface receptors assessed by flow cytometry:
- Relative and absolute change from baseline of mean fluorescence intensity (MFI) and the proportion of positive cells regarding CD25, HLA-DR, LFA-1, CD29, CD69, CD71 expression
• Expression of co-inhibitory molecules assessed by flow cytometry:
- Relative and absolute change from baseline of MFI and of the proportion of positive cells regarding PD-1 = CD279, ICOS = CD278, TIM-3, CTLA4 expression
• Effector functions of CD4 and CD8 positive T cells:
- Relative and absolute change from baseline of the results of cell proliferation assays assessed as the percentage of proliferated cells
- Relative and absolute change from baseline of cytokine production measurement assessed as concentration
- Relative and absolute change from baseline of the cytolytic activity assessed by flow cytometry measurement of MFI and proportion of positive cells regarding Granzyme B, Perforin and CD107a expression
- Relative and absolute change from baseline of intracellular calcium response assessed as a ratio
• Migrational capacity:
- Relative and absolute change from baseline MFI and the proportion of positive cells assessed by flow cytometry expression analysis of CD11a, CD31, CD44, CD49d, CCR5, CCR6, CCR7
- Absolute and relative change of cell numbers of migrated cells compared to baseline assessed in an in vitro model by flow cytometry analysis
• High-throughput TCR- and BCR sequencing of the T- and B cell rep-ertoire concerning the expansion of distinct clones:
- Qualitative comparison of the distribution of CDR3 sequences
• Regulatory T-cell function:
- Relative and absolute change from baseline in production of TGF-beta and IL-10 of CD4+CD25+FOXP3+ regulatory T cells
- Suppression of T cell proliferation: Relative and absolute change from baseline in responder T cell proliferation assessed by suppression assays