Immuno-engineered organoids for regulating the kinetics of B-cell development and antibody production

Developers

Alberto Purwada and Ankur Singh.

Description of the technology

Induction of B-cell immunity against infection depends on the initiation of the germinal center (GC) reaction in secondary lymphoid organs. Ex vivo recapitulation of the GC reaction in 2D cultures results in transient cell growth, with poor yield and short-term survival. Furthermore, no reported 2D ex vivo system can modulate the kinetics of a GC-like phenotype or the rate of antibody class switching.

This protocol, proposed by this technology, describes a methodology for developing immune organoids that partially mimic the B-cell zone of a lymphoid tissue, for efficient and rapid generation of B cells with a GC-like phenotype from naive murine B cells. The organoid is composed of a bioadhesive protein, gelatin, that is transformed into an ionically cross-linked hydrated network using biocompatible silicate nanoparticles (SiNPs). The protocol explains how to establish the immune organoid culture to sustain immune cell proliferation and transformation into a GC-like phenotype. Starting with cell encapsulation in digested lymphoid tissues, clusters of proliferating B cells with a GC-like phenotype can be generated in the organoids at controlled rates, within ~1 week. The culture methodology described in the protocol is currently the only one that allows the accelerated induction of a GC-like phenotype in B cells and supports a controllable immunoglobulin class-switching reaction. This technology can be easily implemented in a typical tissue culture room by personnel with standard mammalian cell culture expertise.

Practical application

This technology has a high potential for the use in both fundamental investigations of immunity and immunotherapy. The engineering of an ex vivo immune organoid according to this technology gives the possibility to overcome the limitations of using live-animal models, 2D systems, and existing 3D models. The immune organoids will enable researchers to reproduce immunological events with tunable parameters to better understand mechanisms of GC-like B-cell phenotype development and antibody class switching, as well as factors that may accelerate malignant lymphoid transformations and differentiation of B-cells into a GC-like B-cell phenotype. At the same time, the 3D organoids may enable studies that will aid in understanding the spatial localization of B cells undergoing controlled differentiation ex vivo in the presence of CD40L and other factors. In the long term immune organoids will grant researchers the ability to identify factors that have the potential to enhance antibody responses against diseases, mechanistic understanding of B-cell hematological malignancies, and immunity in the elderly, and that will allow developing the adequate methods of immunotherapy.

Laboratories

• Meinig School of Biomedical Engineering, Cornell University, Ithaca (USA)
• Sibley School of Mechanical and Aerospace Engineering, Cornell University, Ithaca (USA)

Links

Publications

• Purwada, A. and Singh, A. «Immuno-engineered organoids for regulating the kinetics of B-cell development and antibody production." 12 Nature Protocols (2017): 168–182.
• Purwada, A. et al. «Self-assembly protein nanogels for safer cancer immunotherapy." 5 Adv. Healthc. Mater. (2016): 1413–1419.
• Purwada, A. et al. «Ex vivo engineered immune organoids for controlled germinal center reactions." 63 Biomaterials (2015): 24–34.