'AAV'
Conditions
Interventions
None listed
Sponsors
Universiteit Maastricht
Eligibility
Age
18 Years to 99 Years
Inclusion criteria
Inclusion criteria: Patients: - Patient of the Maastricht University Medical Center (MUMC+) in Maastricht, The Netherlands, or of the Zuyderland Medical Center in Sittard-Geleen, The Netherlands - Diagnosis of respective disease (MS, SLE, AAV, or IBD) - Disease duration
Exclusion criteria
Exclusion criteria: Patients: - Proven infections two weeks prior to blood sampling - Strong immunomodulation and/or immunosuppression in the last 3 months - Other diseases: cancer (malignancies) - Pregnancy;Healthy Controls: - Proven infection two weeks prior to blood sampling - Immune-related diseases - Other diseases: cancer (malignancies) - Pregnancy
Design outcomes
Primary
| Measure | Time frame |
|---|---|
| The primary outcomes of the current study are the following: First, we would like to assess the degree of in vitro proliferation of MACS-isolated regulatory T cells, obtained from patients with autoimmune or inflammatory disease as specified in the hypothesis, in response to the currently applied ex vivo expansion protocol, i.e. interleukin (IL)-2, monoclonal antibodies anti-CD3 and anti-CD28, rapamycin, and the monoclonal anti-TNFR2 antibody MR2-1. The degree of proliferation of Tregs from healthy blood donors will serve as the standard to which the proliferation of Tregs from patients with autoimmune disease will be compared. Second, the suppressive capacity of the expanded Tregs will be analyzed by means of a suppression assay, in which different ratios of Tregs and responder T cells are incubated together and the degree of proliferation of the responder cells is measured. The suppressive capacity of the Tregs can then be expressed as a function dependent on the ratio of Tregs to responder cells (dose-response curve). This function will facilitate the identification of the effective dose (ED) necessary to inhibit 50% of the responder cells from proliferating (ED50). Furthermore, the supernatant of the suppression assay will be screened for the presence of cytokines. | — |
Secondary
| Measure | Time frame |
|---|---|
| The secondary outcomes of the current study are the following: The phenotype (intra-/extracellular) of the Tregs will be analyzed in order to describe the Treg population obtained from the diseased individual and to determine the degree of homogeneity before and after expansion. In order to do so, several characteristic surface and intracellular markers will be investigated. Furthermore, the stability of the expanded Tregs will be assessed by (i) exposing the expanded cells to an artificial pro-inflammatory environment and tracking the cytokine secretion, (ii) analyzing the degree of demethylation of the DNA region *T-Cell Specific Demethylated Region* (TSDR), and (iii) evaluation of expression of characteristic, effector T-cell-associated transcription factors in response to exposure to a pro-inflammatory environment. | — |
Countries
Netherlands
Outcome results
None listed