权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Development & Characterization of Tyrosinase Epitope Specific TCR Transgenic Mice

发展

基本信息

批准号：
7808041
负责人：
Shikhar Mehrotra
金额：
$ 16.43万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-18 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7808041
关键词：
Address Affinity Antigens Autoantigens Autologous Avidity Belief Breeding CD4 Positive T Lymphocytes CD8B1 gene Cell Therapy Cells Cellular biology Clinical Cytotoxic T-Lymphocytes Data Development Enzymes Epitopes Exhibits Frequencies Future Goals Growth HLA-A2 Antigen Human Immunotherapy In Vitro MHC Class I Genes Major Histocompatibility Complex Malignant Neoplasms Mature T-Lymphocyte Melanoma Cell Metastatic Melanoma Monophenol Monooxygenase Mus Mutate Names Patients Peptides Peripheral Physiologic pulse Pigmentation physiologic function Proteins Protocols documentation Reporting Role Self Tolerance Shapes Signal Transduction Source T cell response T-Cell Development T-Cell Receptor T-Lymphocyte T-Lymphocyte Subsets TCR Activation Testing Transgenic Mice Transgenic Organisms Tumor Antigens Tumor-Infiltrating Lymphocytes Vaccination Vaccines base cellular engineering clinically relevant design flexibility human tissue improved in vivo melanocyte melanoma mouse model peripheral tolerance public health relevance receptor receptor expression receptor-mediated signaling response tumor

项目摘要

DESCRIPTION (provided by applicant): Peptide epitopes derived from non-mutated differentiation proteins that are widely expressed by cells of the melanocyte lineage and naturally presented by class I major histocompatibility complex molecules provide targets for vaccination-induced, cytotoxic T lymphocyte (CTL)-based immunotherapy of malignant melanoma. One of the key enzymes involved in pigmentation is the tyrosinase (Tyr) molecule that is expressed not only by malignant melanomas and normal melanocytes, but also by a broad range of human tissues as well. Available data for anti-self human Tyr (hTyr)-based clinical vaccination trials however shows that limited responses were observed in melanoma patients probably due Tyr-specific self-tolerance on the T cell repertoire limited the frequency and avidity of Tyr- reactive CTL. Since T cell receptor (TCR) transgenic mice have contributed to many aspects in understanding T cell ontogeny and selection of the peripheral repertoire, we propose to develop a TCR transgenic mice using a tyrosinase TCR that was previously isolated from a MHC class I- restricted CD4+ T cell isolated from the tumor infiltrating lymphocytes (TILs) of a HLA-A2+ patient with metastatic melanoma. This particular TCR (referred as TIL1383i) has been well characterized and is a co-receptor independent high affinity TCR. The avidity of TIL 1383i for peptide pulsed targets is 10-100-fold lower than most melanoma-reactive CD8+ T cell clones. In our first submission we hypothesized that owing to its origin and high affinity, this TIL1383i TCR may get selected again on a CD4 cell. Indeed we were able to successfully generate the TIL1383i TCR transgenic and our hypothesis proved to be partially correct when we got TIL1383i TCR expression on both CD4 and CD8 T cells. Surprisingly however the T cells passed thymic selection in mice with C57BL/6J background. Now we propose to carry out comprehensive analysis of this new transgenic mouse in context of tumor immunotherapy with the following aims: 1). To characterize antigen recognition and the functional avidity of CD4+ and CD8+ T cells from TIL1383i TCR mice. 2). To determine if CD4+ and CD8+ T cells in TIL1383i TCR mice can control the growth of B16/A2/Kb tumors. We believe that the T cells obtained from the proposed TCR transgenic mice would closely mimic the human T cells and thus the results obtained from the proposed study would help improve us in designing future immunotherapy trials using this unique TCR. PUBLIC HEALTH RELEVANCE: The goal of this project is to develop and characterize a transgenic mice bearing high affinity T cell receptor (TCR) reactive to tumor associated epitope tyrosinase. The high affinity TCR that was used to develop this TCR transgenic mice was isolated from a unique class-I restricted CD4 cell expanded from tumor infiltrating lymphocytes (TILs) of an HLA-A2+ patient with metastatic melanoma (thus referred as TIL1383i TCR). The unusual origin of the TCR and the unique founder obtained on C57BL/6J mice exhibiting TIL1383i TCR expression on both CD4 and CD8 T cell subsets makes this proposal unique since we now propose to understand the role of TCR affinity in tumor response by altering the signal strength of the TIL1383i TCR. This objective will be accomplished by breeding our TIL1383i/Tg mice with commercially available HLA-A2 and HLA-A/H2-Db mice. Successful completion of this proposal would provide us information about the feasibility of using this TIL1383i TCR in T cell adoptive immunotherapy protocols.

描述（由申请人提供）：衍生自非突变分化蛋白的肽表位为恶性黑色素瘤的疫苗诱导的基于细胞毒性T淋巴细胞（CTL）的免疫疗法提供了靶点，所述非突变分化蛋白由黑素细胞谱系的细胞广泛表达并由I类主要组织相容性复合物分子天然呈递。参与色素沉着的关键酶之一是酪氨酸酶（Tyr）分子，其不仅由恶性黑色素瘤和正常黑色素细胞表达，而且也由广泛的人类组织表达。然而，基于抗自身人Tyr（hTyr）的临床疫苗接种试验的可用数据显示，在黑素瘤患者中观察到有限的应答，可能是由于T细胞库上的Tyr特异性自身耐受性限制了Tyr反应性CTL的频率和亲合力。由于T细胞受体（TCR）转基因小鼠在理解T细胞个体发育和外周库选择方面做出了贡献，我们提出使用酪氨酸酶TCR开发TCR转基因小鼠，所述酪氨酸酶TCR先前从MHC I类限制性CD 4 + T细胞分离，所述MHC I类限制性CD 4 + T细胞从患有转移性黑素瘤的HLA-A2+患者的肿瘤浸润淋巴细胞（TIL）分离。这种特定的TCR（称为TIL 1383 i）已被充分表征，并且是共受体非依赖性高亲和力TCR。TIL 1383 i对肽脉冲靶的亲合力比大多数黑素瘤反应性CD 8 + T细胞克隆低10-100倍。在我们的第一次提交中，我们假设由于其来源和高亲和力，该TIL 1383 i TCR可以在CD 4细胞上再次被选择。事实上，我们能够成功地产生TIL 1383 i TCR转基因，并且当我们在CD 4和CD 8 T细胞上都得到TIL 1383 i TCR表达时，我们的假设被证明是部分正确的。然而，令人惊讶的是，T细胞在具有C57 BL/6 J背景的小鼠中通过了胸腺选择。现在，我们建议在肿瘤免疫治疗的背景下对这种新的转基因小鼠进行全面分析，目的如下：1）。表征来自TIL 1383 i TCR小鼠的CD 4+和CD 8 + T细胞的抗原识别和功能性亲合力。2）。确定TIL 1383 i TCR小鼠中的CD 4+和CD 8 + T细胞是否可以控制B16/A2/Kb肿瘤的生长。我们相信，从所提出的TCR转基因小鼠中获得的T细胞将密切模仿人类T细胞，因此从所提出的研究中获得的结果将有助于我们设计未来使用这种独特TCR的免疫治疗试验。公共卫生相关性：本研究的目的是建立一种能与肿瘤相关表位酪氨酸酶反应的高亲和力T细胞受体（TCR）转基因小鼠。用于开发该TCR转基因小鼠的高亲和力TCR分离自从患有转移性黑素瘤的HLA-A2+患者的肿瘤浸润淋巴细胞（TIL）扩增的独特的I类限制性CD 4细胞（因此称为TIL 1383 i TCR）。TCR的不寻常起源和在CD 4和CD 8 T细胞亚群上均表现出TIL 1383 i TCR表达的C57 BL/6 J小鼠上获得的独特建立者使得该提议是独特的，因为我们现在提出通过改变TIL 1383 i TCR的信号强度来理解TCR亲和力在肿瘤应答中的作用。这一目标将通过将我们的TIL 1383 i/Tg小鼠与市售的HLA-A2和HLA-A/H2-Db小鼠交配来实现。该提案的成功完成将为我们提供关于在T细胞过继免疫治疗方案中使用该TIL 1383 i TCR的可行性的信息。