Treg to Th17 conversion in the dynamics of inflammation of the human skin.

Recruitment & Eligibility

Status: Completed

Sex: Not specified

Target Recruitment: 30

Inclusion Criteria

Healthy volunteers must meet the following criteria:
• Adults older than 18 years of age
• Volunteers must be willing to give a written informed consent
• Volunteers must be able to adhere to the visit schedule;Patients must meet the following criteria:
• Adults older than 18 years of age
• All patients have to have actinic keratosis
• The doctor and the patient decided that the best treatment option is field-treatment with imiquimod 5%
• Patients must be willing to give a written informed consent
• Patients must be able to adhere to the visit schedule

Exclusion Criteria

Volunteers will be excluded from this study when any of the following criteria listed below are met:
• Children or adolescents younger than 18 years of age
• Volunteers with a history or signs of psoriasis
• Volunteers with a history or signs of other inflammatory skin diseases, for example atopic dermatitis
• Volunteers exposed to large amounts of sunlight or UV-radiation in the last week
• Volunteers using immunosuppressive agents like prednisone or methotrexate
• Volunteers with relevant co-morbidities
• Volunteers with a current condition involving an activated immune system, such as the flue or a recent vaccination;Patients will be excluded from this study when any of the following criteria listed below are met:
• Children or adolescents younger than 18 years of age
• Patients with a history or signs of psoriasis
• Patients with a history or signs of other inflammatory skin diseases, for example
atopic dermatitis
• Patients exposed to large amounts of sunlight or UV-radiation in the last week
• Patients using immunosuppressive agents like prednisone or methotrexate
• Patients with relevant co-morbidities
• Patients with a current condition involving an activated immune system, such as the
flue or a recent vaccination

Study & Design

Study Type: Observational invasive

Study Design: Not specified

Primary Outcome Measures

Name	Time	Method
<p>The main study endpoint will be the presence of IL-17 producing Tregs in the<br /><br>tissues obtained from healthy volunteers (after tape-stripping and LTB4<br /><br>application) and patients with actinic keratosis treated with imiquimod 5%.<br /><br>Tissues will be analyzed using multicolour immunohistochemistry and<br /><br>immunofleorescence. We developed a sensitive triple staining that is based on<br /><br>the detection of CD4 and Foxp3 expression and IL-17 production. Co-localization<br /><br>of surface differentiation markers, transcription factors and cytokines will be<br /><br>verified by confocal-microscopy. Tissues will be visualized using microscopy,<br /><br>photographed and analysed using computer software (Image J).</p><br>

Secondary Outcome Measures

Name	Time	Method
<p>We will use the mentioned in vivo models for human skin inflammation to analyze<br /><br>the balance of master transcription factors Foxp3 and RORγt through<br /><br>immunohistochemistry and immunofluorescence. We will study co-localization and<br /><br>expression of Foxp3, RORγt and IL-17 by confocal microscopy. </p><br>