Assessment of immunogenicity and antigenicity of different human cell types in natural and 3D-printed allografts

评估天然和 3D 打印同种异体移植物中不同人类细胞类型的免疫原性和抗原性

基本信息

批准号：
10353416
负责人：
JORDAN S POBER
金额：
$ 20.94万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-16 至 2024-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10353416
关键词：
3-Dimensional 3D Print Acute Address Adoptive Transfer Adult Alloantigen Allogenic Allografting Anatomy Animal Model Antigens Biological Assay Biological Models Biomedical Engineering Blood Cells Blood Vessels CRISPR/Cas technology Cell Line Cell Transplantation Cells Clinical Complement Complex Dendritic Cells Dermal Development Dose Drug Delivery Systems Drug Side Effects Effector Cell Endothelial Cells Endothelium Epithelial Cells Fibroblasts Genes Genetic Engineering Goals Graft Rejection Hematopoietic stem cells Human Imaging technology Immune Immune response Immune system Immunity Immunofluorescence Immunologic Immunologics Immunologist In Situ Hybridization Individual Injury Interleukin-2 Intervention Laboratories Left Leukocytes Longevity Mediating Memory Mesenchymal Methods Morphology Mus Myeloid Cells Natural Killer Cells Organ Organ Transplantation Pathway interactions Peptide/MHC Complex Peptides Pericytes Pharmacology Polymers Population Process Property Regulatory T-Lymphocyte Research Support Rest Risk Rodent Role Scientist Signal Transduction Sirolimus Skin Skin graft Stromal Cells T-Lymphocyte Technology Testing Therapeutic Tissue Engineering Tissues Transplantation Umbilical Cord Blood Ursidae Family Work allograft rejection allotransplant anergy cell killing cell type cytokine design dimensional analysis effector T cell high dimensionality human tissue immunogenic immunogenicity improved insight isoimmunity keratinocyte monocyte mouse model nanomedicine nanoparticle neonate new technology penis foreskin progenitor protein complex response success tool uptake

项目摘要

7. PROJECT SUMMARY/ABSTRACT Allograft rejection remains an issue in long term success of allotransplantation and is also likely to negatively impact the function and longevity of tissue engineered grafts being explored as a possible solution to the shortage of available organs. In the direct pathway of allorecognition, rejection is typically initiated by activation of resting host T effector memory cells that cross-react with non-self MHC/peptide complexes, a process that requires graft cells to provide additional signals such as co-stimulators and cytokines. Graft cells capable of doing this are “immunogenic”. Such T cells then differentiate into effector cells that are capable of killing cells that express the same non-self MHC/peptide complexes. Graft cell types that express the same alloantigens and can be recognized and killed by effector T cells but are unable to initiate the differentiation of resting effector memory into effector T cells are said to be “antigenic” rather than immunogenic. Two immunogenic human cell types implicated in initiating rejection are graft dendritic cells (DCs) and graft endothelial cells (ECs). While DCs (and other passenger leukocytes) can be purged from a natural graft or left out of a bioengineered graft, ECs are essential for lining the blood vessels that sustain graft viability. We have proposed strategies to reduce rejection by limiting the immunogenicity of the graft to complement the current practice of suppressing the host immune response. It is unclear if removing DCs and altering ECs to be non- immunogenic will be sufficient to protect a graft from rejection. Here we propose to answer this question. Addressing this question requires that it be performed with human ECs because commonly used rodent ECs do not exhibit the same immunogenic capabilities as their human counterparts. To do so, we will bring to bear 5 novel technologies: a). genetic engineering of ECs to render them non-immunogenic and non-antigenic; b). nanomedicine to modulate the immunogenic capacity of ECs; c). 3D printing of a human skin composed of multiple different cell types as a target for rejection; d). a state-of-the-art human immune system mouse as a graft recipient; and e). cutting edge high dimensional serial immunofluorescence to determine the effects of our interventions on the rejection process. We will initially test our model mouse model and optimize our assays using natural human skin (aim 1) and then proceed to test our 3D skin constructs (aim 2) in which we will modify cell types to determine the role, if any, of human cells other than ECs and DCs. We acknowledge that there is some risk in a proposal that merges multiple technologies which have not been previously combined and therefore have chosen to use the R21 exploratory mechanism to support this research. However, the successful conduct of this project will not only provide an initial answer to an important question for human skin cells, but will also create a platform for further studies of allotransplantation with more complex human tissues and provide valuable insights for bioengineering of replacement organs.

7。项目摘要/摘要在长期的同倍移植成功中，Alllograft拒绝仍然是一个问题，也可能会负面影响探索组织工程移植物的功能和寿命，作为可能的解决方案可用器官短缺。在同种识别的直接途径中，通常通过激活来启动拒绝与非自我MHC/肽复合物交叉反应的静止宿主T效应记忆细胞的需要移植细胞提供其他信号，例如共刺激剂和细胞因子。能够这样做是“免疫原性的”。然后，这样的T细胞分化为能够杀死细胞的效应细胞表达相同的非自我MHC/肽复合物。表达相同同种抗原的移植细胞类型并且可以被效应T细胞识别和杀死，但无法引发静止的分化效应记忆中的效应T细胞被认为是“抗原”，而不是免疫原性。两个免疫原性启动排斥反应实施的人类细胞类型是移植树突状细胞（DC）和移植物内皮细胞（EC）。虽然可以从天然移植物中清除DC（和其他乘客白细胞）生物工程移植物，ECS对于维持移植能力的血管内是必不可少的。我们有提出的策略通过限制移植物的免疫原性来减少排斥的策略抑制宿主免疫反应的实践。目前尚不清楚是否删除DC并将EC改变为非 - 免疫原性足以保护移植物免于排斥。在这里，我们建议回答这个问题。解决这个问题要求它可以与人类EC进行，因为常用的啮齿动物ECS 不要表现出与人类对应物相同的免疫原性能力。为此，我们将带来 5个新技术：A）。 EC的基因工程使它们具有非免疫原性和非抗原性； b）。纳米医学调节EC的免疫原性； c）。由人类皮肤的3D打印多种不同的细胞类型作为拒绝目标； D）。最先进的人类免疫系统鼠标尖端高维连续免疫荧光以确定我们的影响拒绝过程的干预措施。我们最初将测试我们的模型鼠标模型并优化我们的测定使用天然人皮肤（AIM 1），然后继续测试我们的3D皮肤构建体（AIM 2）修改细胞类型以确定EC和DC以外的人类细胞的作用（如果有）。我们承认这一点一项提案中有一些合并多种技术的风险，这些技术以前尚未合并因此，选择使用R21探索机制来支持这项研究。但是，该项目的成功行为不仅将为人类皮肤的重要问题提供初步答案细胞，但还将创建一个平台，用于进一步研究同种异体植入和更复杂的人体组织并为替代组织的生物工程提供宝贵的见解。