Ex vivo generation of murine IL-10-producing B cells by fusokines.

Naïve B cells are crucial components of adaptive immunity. In addition to their capacity to produce immunoglobulins, a minor subset termed regulatory B cells or Bregs has been proven to modulate inflammation through the secretion of soluble mediators. The two main technical difficulties with their clinical use lie in their relatively low abundance in vivo and the scarcity of known methods for their ex vivo expansion. While studying the pharmacological properties of a novel bifunctional granulocyte macrophage-colony-stimulating factor (GM-CSF) and IL-15 fusion transgene (GIFT15) on unfractionated splenocytes in vitro, we observed that the GIFT15 fusokine had a remarkable and unprecedented effect on naïve B cells by converting them into suppressor cells of B-cell ontogeny (hereafter referred to as GIFT15 inducible (i)Bregs). Moreover, GIFT15 promoted iBreg proliferation. We present in this report a detailed protocol using the GIFT15 fusokine as a tool for the ex vivo generation of murine iBregs, which may serve as an immediate remedy to their abundance challenge in the clinic.