In the current *proof of principle* study, it is hypothesized that ex vivo expansion of Tregs originating from patients with the prototypic examples from the spectrum of autoimmune or inflammatory diseases (SLE, AAV, MS or CD) can be accomplished by…
ID
Source
Brief title
Condition
- Gastrointestinal inflammatory conditions
- Autoimmune disorders
- Demyelinating disorders
Synonym
Research involving
Sponsors and support
Intervention
Outcome measures
Primary outcome
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 outcome
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.
Background summary
In autoimmune and inflammatory disease, current immunosuppressive interventions
focus mainly on the inhibition of the pro-inflammatory response. Unfortunately,
this highly effective approach also entails potentially severe side effects.
The ex vivo expansion and subsequent re-administration of autologous regulatory
T lymphocytes (Tregs) seems promising in the search for an alternative, less
toxic, and more specific therapeutic approach. However, the current ex vivo
expansion protocol has been shown to lead to expanded Tregs that lack
sufficient stability and homogeneity, therefore hindering the transition of
this therapeutic approach to the clinic. Recently, the agonistic monoclonal
anti-TNFR2 antibody MR2-1 has been identified to be an important addition to
the ex vivo expansion protocol due to its capability to enhance ex vivo
proliferation, suppressive capacity, homogeneity, and stability. So far, the
advantageous effects of MR2-1 have only been shown in Tregs from healthy
individuals. Therefore, its full potential needs to be verified in Tregs from
diseased individuals. In the current *proof of principle* study, it is
hypothesized that ex vivo expansion of Tregs originating from patients with
autoimmune or inflammatory disease (systemic lupus erythematosus (SLE),
ANCA-associated vasculitides (AAV), multiple sclerosis (MS) or Crohn*s disease
(CD)) by means of the TNFR2 agonist MR2-1 can be accomplished to the same
extent as seen in Tregs from healthy individuals and will lead to improved
proliferation, highly suppressive capacity, high stability and increased
homogeneity.
Study objective
In the current *proof of principle* study, it is hypothesized that ex vivo
expansion of Tregs originating from patients with the prototypic examples from
the spectrum of autoimmune or inflammatory diseases (SLE, AAV, MS or CD) can be
accomplished by means of the monoclonal TNFR2 agonist MR2-1 to the same extent
as seen in Tregs from healthy individuals. Furthermore, it is expected that the
addition of MR2-1 to the ex vivo expansion protocol will lead to a highly
suppressive capacity, increased stability, and increased homogeneity,
independent of the underlying disease.
Study design
A *proof of principle* study will be performed, in which four prototypic
autoimmune-diseased or immune-dysfunctional study groups will be investigated,
i.e. SLE, AAV, MS, and CD, each consisting of 12 participants (Ntotal = 48
patients). A fifth study group will consist of 12 individuals as healthy
controls. Of each participant, venous blood will be sampled once (100 mL): the
blood sample will serve as the source for (i) regulatory T cells
(CD4+CD25+CD127low) for the ex vivo expansion (duration 7 days) and (ii)
responder T cells required for the suppression assay (duration 4 days)
following the expansion of Tregs. In this suppression assay, the responder T
cells will be co-incubated with the expanded regulatory T cells by which the
suppressive capacity of the expanded regulatory T cells is assessed.
Furthermore, the regulatory T cells will be analyzed according to the secondary
outcomes. Other outcomes will consist of analysis of blood values (CRP, ESR,
WBC, vitamin D status) and description of disease activity status (SLEDAI,
BVAS, EDSS, Montreal classification/SES-CD).
The study duration will approximately be 24 months, considering that (i)
processing and analyzing the regulatory T cells will take 11 days per
participant and (ii) blood from a maximum of 3-4 participants can be processed
simultaneously.
Study burden and risks
Burden: Single time venipuncture for blood sampling (phlebotomy), performed at
the MUMC+/ZMC.
Risks: Venipuncture is associated with a low risk of bruising and pain.
Universiteitssingel 50
Maastricht 6229ER
NL
Universiteitssingel 50
Maastricht 6229ER
NL
Listed location countries
Age
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 <5 years
- Ethnicity: Caucasian
- Untreated at the time of blood sampling or prednisone treatment of <10 mg/day
- Age: 18 * 65 years;Healthy Controls:
- Untreated at the time of blood sampling
- Ethnicity: Caucasian
- Age: 18 * 65 years
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
Recruitment
Followed up by the following (possibly more current) registration
No registrations found.
Other (possibly less up-to-date) registrations in this register
No registrations found.
In other registers
| Register | ID |
|---|---|
| CCMO | NL59692.068.16 |