Synthesis and Characterization of Ceramide-Containing Liposomes as Membrane Models for Different T Cell...

A fine balance of regulatory (T_reg) and conventional CD4⁺ T cells (T_conv) is required to prevent harmful immune responses, while at the same time ensuring the development of protective immunity against pathogens. As for many cellular processes, sphingolipid metabolism also crucially modulates the T_reg/T_conv balance. However, our understanding of how sphingolipid metabolism is involved in T cell biology is still evolving and a better characterization of the tools at hand is required to advance the field. Therefore, we established a reductionist liposomal membrane model system to imitate the plasma membrane of mouse T_reg and T_conv with regards to their ceramide content. We found that the capacity of membranes to incorporate externally added azide-functionalized ceramide positively correlated with the ceramide content of the liposomes. Moreover, we studied the impact of the different liposomal preparations on primary mouse splenocytes in vitro. The addition of liposomes to resting, but not activated, splenocytes maintained viability with liposomes containing high amounts of C₁₆-ceramide being most efficient. Our data thus suggest that differences in ceramide post-incorporation into T_reg and T_conv reflect differences in the ceramide content of cellular membranes.