Most antigen discovery and vaccine development aimed at driving functional B cell responses rely on mouse immunizations studies. To date, there is no 3D ex vivo immune 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 apoptosis ex vivo. We have previously reported immune tissues that show ex vivo GC-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 control ex vivo GC reaction. Using an Ezh2fl/fl Cγ1-cre transgenic mouse model, we demonstrated ex vivo IgG1 antibody class switching. Using immune tissues developed from a B1-8hi mutant 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. The ex vivo antigen-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.

中文翻译：

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具有 T 细胞信号的离体合成免疫组织，用于分化抗原特异性、高亲和力生发中心 B 细胞。

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。迄今为止，还没有3D离体免疫组织能够驱动抗原特异性B细胞反应，以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，并产生高亲和力抗体反应。这可归因于生发中心 (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易于离体快速凋亡。我们之前曾报道过显示出离体 GC 样反应的免疫组织，但是以非抗原特异性的方式。在此，我们报道了一种基于马来酰亚胺 (MAL) 功能化聚乙二醇 (PEG) 的设计免疫组织，该组织可在 T 细胞样信号存在的情况下调节 B 细胞分化并富集抗原特异性 GC B 细胞。利用 3D 合成免疫组织平台，我们评估了各种水凝胶设计参数以控制离体 GC 反应。使用 Ezh2fl/fl Cγ1-cre 转基因小鼠模型，我们演示了离体 IgG1 抗体类别转换。使用从 B1-8hi 突变小鼠开发的免疫组织（代表源自 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抗原选择性富集。特异性 B 细胞在细胞表面具有高亲和力，并以整合素配体依赖性方式分泌水平。离体抗原特异性平台技术可用于免疫生物学、基质免疫学和生物技术应用的科学理解，包括抗原测试、疫苗开发和抗疾病抗体的产生。