CytoKine measurement in Skin WaShfluid

Exposure to air pollution, such as ozone (O3), particulate matter (PM), carbon
monoxide (CO), sulfur dioxide (SO2), and nitrogen dioxide (NO2) is strongly
associated with development of diseases of the respiratory tract, the skin and
the eyes. We aim to characterize immune mediators in healthy and inflamed skin,
to identify immune-related biomarkers of interest to study in the context of
air pollution. We propose to develop a system that enables measurement of
immune mediators in skin in a non-invasive manner. As a prove of principle, we
chose to focus first on three pathological skin conditions that differ in
underlying immune pathogenesis and associated cytokines: 1) atopic dermatitis
(AD), a Th2-driven condition; 2) psoriasis, a Th17-driven condition and 3)
cutaneous SLE (CLE), a Th1-skewed interferon-driven disease and healthy
controls. Currently used methods to measure biomarkers in the skin include tape
stripping, and collecting suction blister fluid, both invasive methods. We
propose to use a customized skin wash device to non-invasively collect
biomarkers from skin in a reproducible manner.

Upregulation of the inflammatory response in skin of patients with atopic
dermatitis, psoriasis or cutaneous LE coincides with the expression and
secretion of a distinct set of proinflammatory markers, including cytokines,
chemokines and soluble receptors. We aim to develop a non-invasive method to
collect skin immune mediators.

Primary objectives:
1. To provide proof-of-principle of the skin wash system using recombinant
protein.
2. To identify biomarkers in skin of patients with atopic dermatitis, psoriasis
and cutaneous LE using our skin wash system.

1. To provide proof-of-principle of the skin wash system using recombinant
protein.
To show that we can successfully elute biomarkers of interest from the skin of
healthy controls, we will apply recombinant cytokine solution (IL-1β, IL-6,
IL-8, IL-10, IL-18 and TNFα) to the forearm skin of healthy control volunteers
(n=5). The solution will be allowed to dry, before placing the skin washer: the
well is attached to the arm using an adjustable bracelet. Cold PBS/0.05% T-20
containing the capture beads is added for 30 minutes and the well is closed
with a screw cap. Wash solution will be collected after 30 minutes and analysed
for the cytokines with Luminex technology. Only if we can successfully detect
the recombinant cytokines we move to objective 2.


2. To identify biomarkers in non-invasive skin washes of patients with atopic
dermatitis, psoriasis and cutaneous LE
We aim to include n=10 healthy controls, n=10 atopic dermatitis patients, n=10
psoriasis patients and n=10 cutaneous LE patients.
Patients and healthy controls will be asked to wear the skin wash device on
lesional skin (for patients) and on non lesional skin (both patients and
healthy contols), to demonstrate specific patterns of immune mediator
expression associated with AD. Psoriasis and CLE.
As described before, the well is attached to the arm using an adjustable
bracelet. Cold PBS/0.05% T-20 containing the capture beads is added for 30
minutes and the well is closed with a screw cap. Cytokine levels are determined
with luminex technology.

Participants will wear our skin wash device for 30 minutes, there is no risk
associated with wearing it, and no discomfort is expected.
Although we do not expect this, mild and transient skin irritation might be
caused by the recombinant protein. If this occurs, the even twill be reported.