Immune profiling by high throughput sequencing of complete antigen receptors

适应性免疫系统有能力对几乎无限的病原体和环境威胁做出反应。通过T和B淋巴细胞中抗原受体基因片段的重排和组合缔合，产生了识别这些不同靶标所需的广泛分子多样性。这些可变基因片段的DNA序列决定编码的异源二聚体T细胞受体和B细胞免疫球蛋白的抗原特异性。尽管免疫应答并不完全由特异性决定，但抗原受体作为淋巴细胞识别和接合的主要位点，直接将这组复杂的遗传信息与宿主防御、疫苗接种和自身免疫中的功能联系起来。因此，免疫球蛋白和T细胞受体库的全面序列清单是理解，也许操纵适应性免疫应答的关键一步。然而，确定复杂淋巴细胞群体内不同抗原受体基因的不同序列提出了许多挑战，最显著的是在获得有助于异二聚体抗原受体特异性的两个亚基链的配对序列信息方面。本申请的目标是开发一种高通量策略，其克服了这些挑战并能够进行成对抗原受体亚基的序列分析。结合分子生物学、微流控技术和高通量测序，我将建立一种快速分析T细胞受体和B细胞免疫球蛋白序列的有效方法。这种方法将用于稳态和抗原攻击反应中淋巴细胞库的分子和功能研究。它也将被用来调查的贡献，特别是抗原受体特异性自身免疫发病机制的系统性红斑狼疮模型。这些研究将大大促进我们对健康和疾病中适应性免疫系统的理解。此外，该项目中开发的工具将在研究感染，疫苗接种和自身免疫等领域的免疫功能方面具有广泛的实用性。它们还可以适用于在各种生物系统中以单细胞分辨率分析复杂群体

DESCRIPTION (provided by applicant): The adaptive immune system has the capacity to respond to a virtually limitless array of pathogens and environmental threats. The extensive molecular diversity required to recognize such varied targets is generated through the rearrangement and combinatorial association of antigen receptor gene segments in T and B lymphocytes. The DNA sequence of these variable gene segments determines the antigen specificity of encoded heterodimeric T cell receptors and B cell immunoglobulins. Although an immune response is not dictated by specificity alone, the antigen receptor serves as the principal site for lymphocyte recognition and engagement, directly linking this complex set of genetic information to functions in host defense, vaccination, and autoimmunity. Therefore, a comprehensive sequence inventory of the immunoglobulin and T cell receptor repertoire is a key step towards understanding, and perhaps manipulating, the adaptive immune response. However, determining the distinct sequences of diverse antigen receptor genes within a complex lymphocyte population presents a number of challenges, most notably in acquiring paired sequence information for both of the subunit chains that contribute to the specificity of th heterodimeric antigen receptor. The goal of this application is to develop a high throughput strategy that overcomes these challenges and enables sequence analysis of paired antigen receptor subunits. Using a combination of molecular biology, microfluidics technology, and high throughput sequencing, I will establish an effective method for rapidly profiling T cell receptor and B cell immunoglobulin sequences. This approach will be used for molecular and functional studies of the lymphocyte repertoire in the steady-state and in response to antigenic challenge. It will also be used to investigate the contribution of particular antigen receptor specificities t autoimmune pathogenesis in a model of systemic lupus erythematosus. These studies will significantly advance our understanding of the adaptive immune system in health and disease. Furthermore, the tools developed in this project will have broad utility for studying immune function in areas such as infection, vaccination, and autoimmunity. They can also be adapted for profiling complex populations at single cell resolution in a wide variety of biological systems