Figure 1. Protocol Diagram for Culturing Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit
B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit were cultured in complete Mouse B Cell Expansion Medium as described in Directions For Use (steps 1 - 7). B Cells were harvested on day 9 for analysis. Cells can also be harvested at earlier time points depending on different applications.
Figure 2. Expansion of Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit
B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit were cultured as described in Figure 1. Fold expansion of viable cells is shown with bar graphs representing the mean ± SEM (n = 8). B cells expanded 176.9 ± 29.8-fold after 9 days of culture.
Figure 3. Maturation of Mouse B Cells with ImmunoCult™ Mouse B Cell Expansion Kit
B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit were cultured as described in Figure 1. Following staining using the protocol by Pracht et al. (Eur J Immunol, 2017), the expression of A) B220 and CD138 and B) TACI (CD267) and CD86 were analyzed by flow cytometry at several time points (data represents mean ± SEM, n = 8). An increase in CD86 cell surface expression indicates B cell activation; a decrease in B220 and an increase in CD138 and TACI cell surface expression indicate maturation of B cells to plasmablasts or plasma cells.
Figure 4. Robust Growth of Mouse B Cells when Cultured with ImmunoCult™ Mouse B Cell Expansion Kit
B cells isolated from mouse spleen using EasySep™ Mouse Pan-B Cell Isolation Kit were cultured as described in Figure 1. Cells were imaged at A) 10X magnification on day 0, B) 10X magnification on day 3, C) 40X magnification on day 6, and D) 40X magnification on day 9.
Mouse B cells engineered to express an anti-HPV antibody elicit anti-tumor T cell responses M. G. Bracha et al.
Frontiers in Immunology 2025 Jul
Abstract
Transplantation of engineered B cells has demonstrated efficacy in HIV disease models. B cell engineering may also be utilized for the treatment of cancer. Recent studies have highlighted that B cell activity is associated with favorable clinical outcomes in oncology. In mice,polyclonal B cells have been shown to elicit anti-cancer responses. As a potential novel cell therapy,we demonstrate that engineering B cells to target a tumor-associated antigen enhances polyclonal anti-tumor responses. We observe that engineered B cells expressing an anti-HPV B cell receptor internalize the antigen,enabling subsequent activation of oncoantigen-specific T cells. Secreted antibodies from engineered B cells form immune complexes,which are taken up by antigen-presenting cells to further promote T cell activation. Engineered B cells hold promise as novel,multi-modal cell therapies and open new avenues in solid tumor targeting.
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