Ex vivo synthetic immune tissues with T cell signals for differentiating antigen-specific, high affinity germinal center B cells.
Ex vivo synthetic immune tissues with T cell signals for differentiating antigen-specific, high affinity germinal center B cells.
复制标题
具有 T 细胞信号的体外合成免疫组织，用于分化抗原特异性、高亲和力生发中心 B 细胞。
DOI：
10.1016/j.biomaterials.2018.06.034
复制
发表时间：
2019
期刊：
影响因子：
14
通讯作者：
Singh,Ankur
中科院分区：
文献类型：
--
作者：
Purwada,Alberto;Shah,ShivemB;Béguelin,Wendy;August,Avery;Melnick,AriM;Singh,Ankur
大多数旨在驱动功能性B细胞反应的抗原发现和疫苗开发都依赖于小鼠免疫研究。到目前为止，还没有3Dex活体免疫组织能够驱动抗原特异性B细胞反应来快速确定抗原的体液免疫原性，了解细胞外基质在体液免疫中的作用，并产生高亲和力的抗体反应。这可能归因于生发中心(GC)反应中B细胞分化和亲和力成熟过程的复杂性，这使得这些高度特化的细胞在体内容易发生快速的凋亡。我们之前曾报道过免疫组织表现出活体GC样反应，然而是以非抗原特异性的方式。在这里，我们报告了一种马来酰亚胺(MAL)功能化的聚乙二醇基(PEG)设计的免疫组织，它在T细胞样信号存在的情况下调节B细胞分化并丰富抗原特异性GC B细胞。利用3D合成免疫组织平台，我们评估了控制VvivoGC反应的各种水凝胶设计参数。利用Ezh2fl/Flcγ1-cre转基因小鼠模型，我们证明了活体IgG1抗体类别的转换。利用AB1-8突变小鼠的免疫组织，代表来自4-羟基-3-硝基苯乙酰(NP)半抗原结合抗体(B1-8)的重组抗体可变区，我们证明了抗原特异性和选择性地以整合素配体依赖的方式在细胞表面和分泌水平上选择性地浓缩抗原特异性B细胞。EX VIVOVO抗原特异性平台技术提供了对免疫生物学、基质免疫学的科学理解，以及从抗原测试、疫苗开发到针对疾病抗体的生成的生物技术应用。
Most antigen discovery and vaccine development aimed at driving functional B cell responses rely on mouse immunizations studies. To date, there is no 3Dex vivoimmune tissues, which are capable of driving antigen-specific B cell responses to rapidly determine the humoral immunogenicity of antigens, understand the role of extracellular matrix in humoral immunity, and generate high affinity antibody responses. This can be attributed to the complexity of B cell differentiation and affinity maturation process in the germinal center (GC) reaction, which makes these highly specialized cells susceptible to rapid apoptosisex vivo. We have previously reported immune tissues that showex vivoGC-like response, however in a non-antigen specific manner. Here, we report a maleimide (MAL)-functionalized polyethylene glycol (PEG)-based designer immune tissues that modulate B cell differentiation and enriches antigen-specific GC B cells in the presence of T-cell like signals. With the 3D synthetic immune tissue platform, we assessed various hydrogel design parameters to controlex vivoGC reaction. Using anEzh2fl/flCγ1-cre transgenic mouse model, we demonstratedex vivoIgG1 antibody class switching. Using immune tissues developed from aB1-8himutant mouse that represents a recombined antibody variable region derived from a 4-hydroxy-3-nitrophenylacetyl (NP) hapten binding antibody (B1-8), we demonstrate antigen specificity and selective enrichment of antigen-specific B cells with high affinity at both cell surface and secreted levels in integrin ligand-dependent manner. Theex vivoantigen-specific platform technology offers use in scientific understanding of immunobiology, matrix immunology, and in biotechnology applications, ranging from the antigen testing, vaccine development, and generation of antibodies against diseases.