Antigen-specific immune reactions to de novo protein expression upon EB therapy (Gratz-Murauer 1)Completed
|Project lead||Assoc. Prof Iris Gratz and Dr Eva Murauer|
|Organisation||EB House Austria, Salzburg, AUSTRIA|
|Project budget||EUR 159,880.00|
|Start date / Duration||01. Mar 2014 / 31 months|
|Funder(s) / Co-Funder(s)||DEBRA Austria, MSAP/EBEP Recommended|
|Research area||Molecular therapy, Cellular therapy, Immunology|
Publications related to the projectsHuman CD4+CD103+ cutaneous resident memory T cells are found in the circulation of healthy individuals Taking the lead – how keratinocytes orchestrate skin T cell immunity A novel humanized mouse model to study human antigen-specific cutaneous T cell responses in vivo Organ-Specific and Memory Treg Cells: Specificity, Development, Function, and Maintenance
Short lay summary
In this project, we developed a novel mouse model to study immune cells of human donors responding to protein in the skin in order to in the future test EB patient’s immune reactions to therapeutic molecules present in the skin. In this model, we transfer human blood-derived immune cells into mice that lack their own immune system (i.e. are immuno-deficient), and in parallel we graft these mice with human skin cells that were cultured in vitro, using a procedure similar to that used in ex vivo gene therapy approaches. By combining immune cells with the human skin tissue, we can test the immune reaction to a protein in the skin (e.g. microbial components or the therapeutic protein such as collagen VII for the therapy of RDEB).
In this work, we aimed to address the immunological consequences of EB therapy. Research to therapeutically treat EB is advancing rapidly and multiple therapies are now on the horizon. Faulty genes which result in the absence or malfunction of specific skin proteins cause EB, and all therapeutic approaches aim to introduce the correct skin protein to EB patients. However, we have learned from EB animal models and various settings of causative therapy that the newly introduced protein can activate the immune system, leading to inflammation and rejection of the therapy. This happens because the patients’ immune cells are unfamiliar with the corrected protein introduced by therapy (protein, gene or cell). Thus, in order to gauge the immune reaction to the specific protein prior to any therapy we wished to develop a model to test immune responses of skin-tropic T cells of any given patient.
- DEBRA strategic goal: several goals of DEBRA Research, particularly molecular and cellular therapies for EB, cancer research and research into skin fibrosis will be promoted by the use of in vivo systems to model human skin disease. Thus, we aim to provide the groundwork and scientific rationales to develop therapies for various symptoms of EB.
- Project goal: to investigate skin immunity by human donors.
- Preceding/ follow-on projects, and related projects: The mouse model developed in this project is now utilized in an NIH funded project (R01AI127726) as well as preclinical cancer therapy research performed in collaboration with GammaDeltaTherapeutics.
What did this project achieve?
Over the course of the project we have achieved the following milestones:
- We have optimized the human immune cell engraftment into recipient mice to give these a human immune system.
- We have established a simple grafting method to generate human skin on recipient mice.
- We have developed methods to follow the migration of immune cells to the human skin graft of recipient mice.
- We have found that human immune cells can react against foreign antigen (such as the fungus Candida albicans) when it is present in the human skin graft.
- We have extended this method to test the reaction against collagen VII in the skin.
- We have established a patient’s sample library that contains blood-immune cells and isolated skin cells from 11 RDEB and 3 DDEB patients, which is a higher number than we had initially proposed.
- We have immortalized RDEB and DDEB keratinocytes to allow virtually unlimited growth in vitro prior to gene correction.
We will now use this mouse model to study the role of special immune cells in the skin, particularly focussing on their function in skin wound healing as well as skin cancer.