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. 项目概要/摘要同种异体移植排斥仍然是同种异体移植长期成功的一个问题，并且也可能产生负面影响影响组织工程移植物的功能和寿命，正在探索作为解决该问题的可能解决方案在同种异体识别的直接途径中，可用器官的短缺，排斥通常是由激活引发的。静息宿主 T 效应记忆细胞与非自身 MHC/肽复合物发生交叉反应，这一过程需要移植细胞提供额外的信号，例如共刺激剂和细胞因子。这样做是“免疫原性的”，然后此类 T 细胞分化成能够杀死细胞的效应细胞。表达相同的非自身 MHC/肽复合物的移植细胞类型，表达相同的同种异体抗原。可以被效应 T 细胞识别并杀死，但无法启动静息 T 细胞的分化效应T细胞的效应记忆被认为是“抗原性的”而不是免疫原性的。参与引发排斥反应的人类细胞类型是移植物树突状细胞 (DC) 和移植物内皮细胞（EC），而 DC（和其他过客白细胞）可以从自然移植物中清除或从移植物中去除。生物工程移植物中，EC 对于维持移植物活力的血管内壁至关重要。提出了通过限制移植物的免疫原性来减少排斥反应的策略，以补充当前的免疫原性目前尚不清楚去除 DC 和改变 EC 是否是非-抑制宿主免疫反应的做法。免疫原性足以保护移植物免受排斥。在这里我们建议回答这个问题。解决这个问题需要用人类 EC 来执行，因为常用的啮齿动物 EC 不表现出与人类捐赠者相同的免疫原性能力为此，我们将采取行动。 5 项新技术：a) 使 EC 具有非免疫原性和非抗原性；纳米药物调节 EC 的免疫原性；c). 多种不同的细胞类型作为排斥目标；d). 移植受体；和e)。我们将首先测试我们的模型小鼠模型并优化我们的测定使用天然人体皮肤（目标 1），然后继续测试我们的 3D 皮肤结构（目标 2），其中我们将修改细胞类型以确定除 EC 和 DC 之外的人类细胞的作用（如果有）。合并先前未合并的多种技术的提案存在一些风险因此选择使用R21探索机制来支持这项研究。该项目的成功实施不仅将为人类皮肤的一个重要问题提供初步答案细胞，但也将为进一步研究更复杂的人体组织的同种异体移植创建一个平台并为替代器官的生物工程提供有价值的见解。