TitleEx vivo synthetic immune tissues with T cell signals for differentiating antigen-specific, high affinity germinal center B cells.
Publication TypeJournal Article
Year of Publication2019
AuthorsPurwada, Alberto, Shah Shivem B., Béguelin Wendy, August Avery, Melnick Ari M., and Singh Ankur
Date Published2019 Apr
KeywordsAnimals, Antigens, B-Lymphocytes, Biocompatible Materials, Cell Differentiation, Cells, Cultured, Female, Germinal Center, Immunoglobulin G, Male, Maleimides, Mice, Inbred C57BL, Polyethylene Glycols, T-Lymphocytes, Tissue Engineering

<p>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 Ezh2 Cγ1-cre transgenic mouse model, we demonstrated ex vivo IgG1 antibody class switching. Using immune tissues developed from a B1-8 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.</p>

Alternate JournalBiomaterials
PubMed ID30041943
PubMed Central IDPMC6355359
Grant ListR35 CA220499 / CA / NCI NIH HHS / United States
R01 CA187109 / CA / NCI NIH HHS / United States
R01 AI132738 / AI / NIAID NIH HHS / United States
S10 RR025502 / RR / NCRR NIH HHS / United States
T32 EB023860 / EB / NIBIB NIH HHS / United States